title: A Survey of County Health Departments of Kansas Regarding COVID-19

text: INTRODUCTION: SARS-CoV-2 virus (severe acute respiratory syndrome coronavirus 2) causing COVID-19 (Coronavirus Disease 2019) initially was identified in China in December 2019. It has resulted in a pandemic with increasing spread of the virus in the U.S. The county health departments around U.S. are spearheading the response to contain the spread of this virus. METHODS: This project was a survey of county health departments in the state of Kansas with data collection period from April 15 to April 24, 2020. This study evaluated the staffing, resources, and funding of these health departments and how it was affecting the efforts to contain COVID-19. Descriptive statistics were used to summarize the responses. RESULTS: A total of 75% of the county health departments in Kansas responded to the survey. In 89% of locations, the staffing had not increased. Most health departments had an average of five people and the four largest ones had 30 to 98 staff working on COVID-19. Most locations used the Kansas Department of Health and Environment criteria for testing and used a combination of state or private laboratories. The results of the tests were available three days or longer in 62% and after five days in 14% of sites. All locations were active in contact tracing, but most had one to three people for this purpose and in 90% the contact tracing interview was via phone calls. There was no change in funding in 21% and decreased funding in 8.5% of health departments. Most locations had an average of five nasopharyngeal swabs on the day of the survey. The most common needs expressed were help to increase testing capability, more public education, more personal protective equipment, increased personnel, and assistance with contract tracing. CONCLUSION: There is an urgent need in Kansas to increase support to county health departments for testing capability, personal protective equipment, increased number of staff, increased help with contact tracing, and especially increase support for public education.


COVID-19: A global public health disaster


COVID-19: Lost opportunities and lessons for the future


COVID-19: Utilizing local experience to suggest optimal global strategies to prevent and control the pandemic


title: Artificial Intelligence-Powered Search Tools and Resources in the Fight Against COVID-19

text: Emerging technologies are set to play an important role in our response to the COVID-19 pandemic. This paper explores three prominent initiatives: COVID-19 focused datasets (e.g., CORD-19); Artificial intelligence-powered search tools (e.g., WellAI, SciSight); and contact tracing based on mobile communication technology. We believe that increasing awareness of these tools will be important in future research into the disease, COVID-19, and the virus, SARS-CoV-2.


COVID-19: Armageddon before Light?


title: Thrombus in Transit and Impending Pulmonary Embolism Detected on POCUS in a Patient with COVID-19 Pneumonia

text: Coronavirus disease 2019 (COVID-19) is a pandemic that started in China in December 2019 and carries a high risk of morbidity and mortality. To-date (4-22-2020) it affected over 2.6 million people and resulted in nearly 200,000 death worldwide mainly due to severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2). Among the major underlying pathophysiologic mechanisms in COVID 19 is hypercoagulability, leading to increased risk for deep vein thrombosis and pulmonary embolism that contribute to increased morbidity and mortality. In this report, we present the case of a 55-year-old man who presented with COVID-19 pneumonia, and was found to have a thrombus in transit by routine point of care ultrasound (POCUS). While computer tomography (CT) angiography is the test of choice, the utilization of point of care ultrasound (POCUS) has gained traction as an adjunctive means of surveillance for the development of VTE in patients with COVID-19. In this report, we discuss the clinical utility of POCUS in diagnosing thrombus in transit in COVID 19 populations.


title: Correlation between Chest Computed Tomography Scan Findings and Mortality of COVID-19 Cases; a Cross sectional Study

text: INTRODUCTION: Predicting the outcomes of COVID-19 cases using different clinical, laboratory, and imaging parameters is one of the most interesting fields of research in this regard. This study aimed to evaluate the correlation between chest computed tomography (CT) scan findings and outcomes of COVID-19 cases. METHODS: This cross sectional study was carried out on confirmed COVID-19 cases with clinical manifestations and chest CT scan findings based on Iran's National Guidelines for defining COVID-19. Baseline and chest CT scan characteristics of patients were investigated and their correlation with mortality was analyzed and reported using SPSS 21.0. RESULTS: 380 patients with the mean age of 53.62 ± 16.66 years were evaluated (66.1% male). The most frequent chest CT scan abnormalities were in peripheral (86.6%) and peribronchovascular interstitium (34.6%), with ground glass pattern (54.1%), and round (53.6%) or linear (46.7%) shape. There was a significant correlation between shape of abnormalities (p = 0.003), CT scan Severity Score (CTSS) (p <0.0001), and pulmonary artery CT diameter (p = 0. 01) with mortality. The mean CTSS of non-survived cases was significantly higher (13.68 ± 4.59 versus 8.72 ± 4.42; <0.0001). The area under the receiver operating characteristic (ROC) curve of CTSS in predicting the patients’ mortality was 0.800 (95% CI: 0.716-0.884). The best cut off point of chest CTSS in this regard was 12 with 75.82% (95% CI: 56.07%-88.98%) sensitivity and 75.78% (95% CI: 70.88%-80.10%) specificity. The mean main pulmonary artery diameter in patients with CTSS ≥ 12 was higher than cases with CTSS < 12 (27.89 ± 3.73 vs 26.24 ± 3.14 mm; p < 0.0001). CONCLUSION: Based on the results of the present study it seems that there is a significant correlation between chest CT scan characteristics and mortality of COVID-19 cases. Patients with lower CTSS, lower pulmonary artery CT diameter, and round shape opacity had lower mortality.


title: Dividing the Emergency Department into Red, Yellow, and Green Zones to Control COVID-19 Infection; a Letter to Editor

text: COVID-19, in certain respects, can be viewed as a CBRN (chemical, biological, radiological, or nuclear) event due to being a consequence of SARS-CoV2 virus (the “contaminant”). We, thus, reorganized our emergency department (ED) into 3 distinct zones (red, yellow, and green) for the purpose of infection control. Patients with high or medium risk of COVID-19 infection are managed in the red zones. Low-risk patients are managed in the yellow zones. All patients are prohibited to enter the green zones. Green zones are used by healthcare providers (HCPs) for personal protective equipment (PPE) donning, inventory, planning, and dining. Only HCPs who work in the red zones are required to use full level PPE (aerosol precaution). HCPs working in the yellow zones require less PPE (contact and droplet precaution). No PPE is required in the green zones. Establishing red, yellow, and green zones in the ED can be helpful in reducing cross-infections and minimizing demand for PPE.


title: Massive Pulmonary Thromboembolism in Patients with COVID-19; Report of Three Cases

text: COVID-19 is a novel infectious disease, which has challenged people all around the world. As of today, healthcare practitioners and researchers have made great effort to understand the characteristics and clinical presentations of the disease; however, the existing literature is still incomplete in this regard. A growing body of evidence indicates that coagulopathies and thromboembolic events are of utmost importance in COVID-19 patients and are related to poor prognosis. Here, we report three ICU admitted cases of COVID-19, in which massive pulmonary thromboembolism (PTE) occurred a few days after disease onset. Unfortunately, one of the patients did not survive and two were treated; one with thrombectomy and other with antithrombotic agents. It seems that severe cases of COVID-19 are at risk for developing PTE and in-charge physicians should be prepared and plan for anticoagulant prophylaxis using low-molecular-weight heparin (LMWH).


Applications of Traditional Chinese Medicine in Antiviral and Anticancer Drug Development: An Interview with Dr. Yung-Chi (Tommy) Cheng, PhD


Ten Drugs: How Plants, Powders, and Pills Have Shaped the History of Medicine


title: Management of ST-Elevation Myocardial Infarction in the COVID-19 Era: The Role of Thrombosis and Anticoagulation Strategy

text: Cardiac manifestations of COVID-19 include myocarditis, demand ischemia, myocardial infarction and arrhythmias with prothrombotic state being a major underlying pathogenetic mechanism. In this report we present a case of a 57-year-old, otherwise healthy, woman who presented with chest pain and nausea and was found to have an inferior wall ST-elevation myocardial infarction (STEMI) in the setting of an active COVID-19 infection. Angiography revealed tortuous coronary arteries with a 100% right coronary artery occlusion with high thrombus burden and normal left coronary system. In light of the available literature regarding the pro-thrombotic effects of this novel corona virus, we continued full dose anticoagulation with Enoxaparin after the cardiac catheterization and transitioned to rivaroxaban and we also continued the patient on dual antiplatelet therapy prior to discharge.


title: Kansans in the Middle of the Pandemic: Risk Perception, Knowledge, Compliance with Preventive Measures, and Primary Sources of Information about COVID-19

text: INTRODUCTION: As we conduct this study, the world is in the grasp of a deadly pandemic. In less than six months since its first diagnosis in Wuhan, China, the COVID-19 infectious disease due to the novel coronavirus has infected over 5,000,000 people and claimed over 350,000 lives. In the United States, most of the cases are in large urban settings along the coasts, but the disease is slowly progressing through the mainland. Kansas, with its particular location in the midwest United States, has seen a relatively small number of cases, but these are increasing. The Kansas government took radical measures to prevent the spread of the disease. According to the Health Beliefs Model, an individual’s perception of risk will dictate engagement with preventive behaviors. Knowledge about the disease and preventive measures drive the risk assessment. Knowledge is dependant on the sources of information used. This study explored these metrics in a sample of Kansans living in the times of the COVID-19 pandemic. METHODS: A combination of snowball samples and random distribution through social media was used to recruit participants to an online survey. The risk and knowledge instrument was developed and validated by WHO Europe. Data collection lasted 96 hours. RESULTS: The attitudes and behaviors of Kansans concerning COVID-19 were consistent with its location in an area of the country with a relatively lower incidence of the disease. Participants had good knowledge about the disease and preventive measures and were willing to comply with recommendations from local authorities. CONCLUSION: Localized information sources that cater to the community are often primary, while social media is not a valuable source for information pertinent to COVID-19.


Statins as early therapy to mitigate COVID-19 (SARS-CoV-2)-associated ARDS and cytokine storm syndrome – time is of the essence


title: Whole Genome Analysis and Targeted Drug Discovery Using Computational Methods and High Throughput Screening Tools for Emerged Novel Coronavirus (2019-nCoV)

text: A novel coronavirus designated as SARS-CoV-2 in February 2020 by World Health organization (WHO) was identified as main cause of SARS like pneumonia cases in Wuhan city in Hubei Province of China at the end of 2019. This been recently declared as Global Pandemic by WHO. There is a global emergency to identify potential drugs to treat the SARS-CoV-2. Currently, there is no specific treatment against the new virus. There is a urgency to identifying potential antiviral agents to combat the disease is urgently needed. An effective and quick approach is to test existing antiviral drugs against. Whole genome analysis and alignment carried out using BLASTn, SMART BLAST and WebDSV 2.0 had shown more than 238 ORF’s coding for proteins mostly origin from Bat SARS coronavirus and root genomic origin from Archaea. Molecular docking results against protein targets Furin, papain like proteases, RdRp and Spike glycoprotein had shown paritaprevir, ritonavir, entecavir and chloroquine derivatives are the best drugs to inhibit multi targets of coronavirus infection including natural compounds corosolic acid, baicalin and glycyrrhizic acid with minimal inhibitory concentrations. Thus we propose use of paritaprevir, entecavir, ritonavir and chloroquine derivatives as best drug combination along with niacinamide, folic acid and zinc supplements to treat novel coronavirus infection. We also propose use of plant protease inhibitors (PI’s) and Anti-IL8, IL-6, IL-2 as future drug models against coronavirus.


Traditional Healing Practices Involving Psychoactive Plants and the Global Mental Health Agenda: Opportunities, Pitfalls, and Challenges in the “Right to Science” Framework


title: Addressing the Problem of Severe Underinvestment in Mental Health and Well-Being from a Human Rights Perspective

text: Throughout the world, mental health remains a neglected priority, low on the agenda of policy makers and funders at the national and international levels. While this is shifting somewhat, there remains a considerable need to address the underprioritization of mental health and well-being, perhaps even more so in the wake of the COVID-19 pandemic. However, given the history of mental health interventions—which have overemphasized the biomedical model and have thus resulted in coercion, denial of life in the community, and unnecessary pathologization of human experience—there is also a need to ensure that increased funding does not simply replicate these mistakes. This is particularly true in the current landscape, where efforts to “scale up” mental health and to reduce “treatment gaps” are gaining momentum and where post-pandemic responses are still being formulated. As the potential for global mechanisms for funding mental health increases, national and international funders should look to practices that are rights affirming and contextually relevant. In this paper, I explore the current landscape of mental health financing, in terms of both national resource allocation and development assistance. I then outline the momentum in global mental health that is likely to materialize through increased funding, before considering ways in which that funding might be utilized in a manner that promotes human rights.


title: A Key, Not a Straitjacket: The Case for Interim Mental Health Legislation Pending Complete Prohibition of Psychiatric Coercion in Accordance with the Convention on the Rights of Persons with Disabilities

text: The practice of coercion on the basis of psychosocial disability is plainly discriminatory. This has resulted in a demand from the Committee on the Rights of Persons with Disabilities (the CRPD Committee) for a paradigm shift away from the traditional biomedical model and a global ban on compulsion in the psychiatric context. However, that has not occurred. This paper considers conflicting pronouncements of the CRPD Committee and other United Nations bodies. Assuming the former’s interpretations of the Convention on the Rights of Persons with Disability (CRPD) are accurate, involuntary psychiatric detention and enforced treatment on the basis of psychosocial disability are prima facie discriminatory and unlawful practices. However, dedicated mental health legislation both permits discrimination and protects and enhances rights. This paper proposes a practical way out of the present impasse: the global introduction of interim “holding” legislation lacking full compliance with the CRPD. While imperfect, such a framework would facilitate a move toward a complete ban on psychiatric coercion. The paper outlines four essential ingredients that any interim legislation ought to contain, including clear timebound targets for full CRPD implementation. It concludes by urging the CRPD Committee to take the unprecedented step of issuing a general comment providing reluctant “permission” for the progressive realization of respect for articles 12 and 14 of the CRPD.


title: Understanding US Immigration Detention: Reaffirming Rights and Addressing Social-Structural Determinants of Health

text: A crisis of mass immigration detention exists in the United States, which is home to the world’s largest immigration detention system. The immigration detention system is legally classified as civil, rather than criminal, and therefore non-punitive. Yet it mimics the criminal incarceration system and holds detained individuals in punitive, prison-like conditions. Within immigration detention centers, there are increasing reports and recognition of civil and human rights abuses, including preventable in-custody deaths. In this paper, we propose understanding the health impacts of detention as an accumulation of mental and physical trauma that take place during the entirety of a detained immigrant’s experience, from migration to potential deportation and removal. Further, we explore the social-structural determinants of health as they relate to immigration detention, contextualize these determinants within a human rights framework, and draw parallels to the larger context of US mass incarceration. Realizing the right to health requires addressing these social-structural determinants of health. For the care of immigrant patients to be effective, clinicians and public health professionals must incorporate an awareness of the health risks of the immigration detention system into trauma- and human rights-informed models of care during and after detention.


COVID-19 and Detention: Respecting Human Rights


title: Global Health in the Age of COVID-19: Responsive Health Systems Through a Right to Health Fund

text: We propose that a Right to Health Capacity Fund (R2HCF) be created as a central institution of a reimagined global health architecture developed in the aftermath of the COVID-19 pandemic. Such a fund would help ensure the strong health systems required to prevent disease outbreaks from becoming devastating global pandemics, while ensuring genuinely universal health coverage that would encompass even the most marginalized populations. The R2HCF’s mission would be to promote inclusive participation, equality, and accountability for advancing the right to health. The fund would focus its resources on civil society organizations, supporting their advocacy and strengthening mechanisms for accountability and participation. We propose an initial annual target of US$500 million for the fund, adjusted based on needs assessments. Such a financing level would be both achievable and transformative, given the limited right to health funding presently and the demonstrated potential of right to health initiatives to strengthen health systems and meet the health needs of marginalized populations—and enable these populations to be treated with dignity. We call for a civil society-led multi-stakeholder process to further conceptualize, and then launch, an R2HCF, helping create a world where, whether during a health emergency or in ordinary times, no one is left behind.


Reimagining the Mental Health Paradigm for Our Collective Well-Being


2019 NOVEL CORONAVIRUS


COVID-19 Pandemic and the New State of Oncology Practice: An Editorial


title: The Fgl2/fibroleukin prothrombinase contributes to immunologically mediated thrombosis in experimental and human viral hepatitis.

text: Fibrin deposition and thrombosis within the microvasculature is now appreciated to play a pivotal role in the hepatocellular injury observed in experimental and human viral hepatitis. Importantly, the pathways by which fibrin generation is elicited in viral hepatitis may be mechanistically distinct from the classical pathways of coagulation induced by mechanical trauma or bacterial lipopolysaccharide (LPS). In the setting of murine hepatitis virus strain-3 (MHV-3) infection, a member of the Coronaviridae, activated endothelial cells and macrophages express distinct cell-surface procoagulants, including a novel prothrombinase, Fgl2/fibroleukin, which are important for both the initiation and localization of fibrin deposition. To assess the role of Fgl2/fibroleukin in murine viral hepatitis we generated a Fgl2/fibroleukin-deficient mouse. Peritoneal macrophages isolated from Fgl2/fibroleukin-/- mice did not generate a procoagulant response when infected with MHV-3. Fibrin deposition and liver necrosis were markedly reduced, and survival was increased in mice infected with MHV-3. To address the relevance of Fgl2/fibroleukin in human chronic viral hepatitis we studied patients with minimal and marked chronic hepatitis B. We detected robust expression of Fgl2/fibroleukin mRNA transcripts and protein in liver tissue isolated from patients with marked chronic hepatitis B. Fibrin deposition was strongly associated with Fgl2/fibroleukin expression. Collectively, these data indicate a critical role for Fgl2/fibroleukin in the pathophysiology of experimental and human viral hepatitis.


WHO issues guidelines to manage any future SARS outbreak.


Fatal outcome after inadvertent injection of topical EPINEPHrine.


title: Winter dysentery in dairy herds: electron microscopic and serological evidence for an association with coronavirus infection.

text: Faeces and, or, paired sera were collected from cows in six dairy herds with classical winter dysentery. Similar samples were collected from cows in three other dairy herds experiencing non-haemorrhagic diarrhoea during the survey period. Coronavirus was the only enteric pathogen identified by immune electron microscopy (IEM) in all six outbreaks, occurring in 26 of 29 (90 per cent) of the affected cows and in one of 11 normal cows from the same herds. Nineteen of 26 affected cows (73 per cent) developed greater than four-fold increases in neutralising antibody titres to the Mebus strain of bovine coronavirus, compared with two of eight normal cows in the same herds. No cows showed greater than four-fold increases in antibody titres to bovine virus diarrhoea virus. None of the cows from the three herds with non-haemorrhagic diarrhoea shed coronavirus in faeces detectable by IEM or developed greater than two-fold rises in coronavirus antibody titres in paired sera. No enteric pathogens were identified in two of the herds. However, two cows in the third herd shed a group B rotavirus detected by IEM. These findings provide additional evidence for a possible role for bovine coronavirus in the aetiology of winter dysentery. Furthermore, this is the first report of a group B rotavirus associated with diarrhoea in adult cattle.


title: Seromonitoring in small laboratory animal colonies. A five year survey: 1984-1988.

text: From 1984 to 1988 one thousand serologic investigations of laboratory animal colonies originating from 10 different European countries were performed. The most prevalent infections in mouse stocks were caused by Mouse hepatitis virus (MHV), Minute virus of mice (MVM), Theiler's encephalomyelitis virus (TEMV), Reovirus type 3 (Reo3), Sendaivirus, and Pneumonia virus of mice (PVM). In mice no infections with Lymphocytic choriomeningitis virus (LCM), Polyomavirus, Mouse adenovirus, and K-virus were recorded. Only two colonies were infected by Ectromelia virus. The first six virus infections of mice were also found in rat colonies as well as the rat-specific Coronaviruses (Sialodacryoadenitisvirus--SDA, Rat corona virus--RCV) and Parvovirus (Kilham rat virus--KRV, Toolan H-1 virus) being endemic. Antibodies to Bacillus piliformis were detectable in about 50% of rat stocks screened. This is in contrast to the mouse, where only about 10% of the colonies were found to be positive. A similar picture was seen for M. pulmonis which is primarily an infection of the rat. In mice no case was detected during the last two years. The number of investigations performed from guineapig, hamster and rabbit colonies was relatively low. Nevertheless, antibodies against the following antigens were detectable: In guineapig stocks: Reo3, PVM, Sendai, Simian virus 5 (SV5) and B. piliformis; in rabbits: Reo3, Sendavirus, SV5, and B. piliformis; in hamsters: PVM, LCM and B. piliformis. The overall contamination rate showed a continuous decrease until 1988. Nevertheless, about 50% of mouse and rat stocks still exhibited antibodies to one or more viral infections.


title: Serological screening of Tick-borne encephalitis (TBE) among Malaysian encephalitis patients.

text: Tick-borne encephalitis (TBE) is a viral infection of the central nervous system and is caused by tick bites, usually after travel to rural or forested areas. The disease is prevalent in Scandinavia, Western Europe, Central Europe and the former Soviet Union and East Asia including Japan. In Malaysia, so far there are no reported cases of TBE. In the present time, many illnesses have been attributed to traveling to other parts of the world. Thus it is important to carry out TBE prevalence study to determine whether the virus is present among Malaysian population. Samples (sera and CSF) from patients admitted to major MOH hospitals in Peninsular Malaysia and Sabah with a clinical diagnosis of encephalitis but is IgM negative for JE, were tested for TBEV IgM ELISA and TBEV IgG ELISA (DRG, Germany). Out of the 600 samples screened for TBEV IgG, all were non-reactive. In addition, out of the 100 samples screened for TBEV IgM, all the samples were also non-reactive. Our results indicate that currently TBE is not present in the Malaysian population. Among the reasons for this could be lack of the infection agent, absence of the suitable vector or subjects selected for the study did not fit the criteria of possible exposure to TBE infections. Hence we recommend that for any future study, the selection of subjects should include those who returned from tick-infested forested areas.


title: Frequent flyer business travelers: major exposure hazards.

text: Bagshaw (2004) notes "the modern commercial aircraft cabin is maintained with adequate environmental control for the comfort of most healthy individuals" (p. 417). Occupational health nurses frequently deal with a population that may include unhealthy individuals or those with pre-existing conditions. It is critical for occupational health nurses to stay current with major hazards faced by frequent flyer business travelers to assist in identifying and preventing adverse health effects associated with these exposures.


title: Duplex ultrasound assisted endovascular revascularization of chronic internal carotid artery occlusion: technical note.

text: Carotid artery occlusion is a major risk factor for ischemic stroke. Endovascular treatment of carotid artery occlusion in high-risk patients offers a therapeutic option to reduce subsequent ischemic events. The technical success and avoidance of complications of the endovascular recanalization of occluded carotid artery is contingent on proper visualization of the vessel wall. To highlight the value of duplex ultrasound guidance, we herein described the procedural details of the use of ultrasound guidance for characterization and visualization of the endovascular devices as well as the vessel wall during endovascular revascularization. Patient was a 54-year-old woman, diagnosed with carotid occlusion with recurrent symptomatic episodes. Endovascular revascularization was successfully performed by ultrasound assistance upon traversing the occluded segment and accurate deployment of stent. The patient was followed for 3 months and has not experienced any ischemic symptoms. Duplex ultrasound guidance helps preventing the vessel wall injury and dissection during revascularization of chronically occluded carotid artery. Ultrasound guidance may improve the outcomes of the endovascular procedures in high-risk patients with carotid occlusion.


title: Outbreaks following wild poliovirus importations --- Europe, Africa, and Asia, January 2009-September 2010.

text: The Global Polio Eradication Initiative (GPEI) began in 1988. By 2006, indigenous transmission of wild poliovirus (WPV) had been interrupted in all but four countries (Afghanistan, India, Nigeria, and Pakistan). However, outbreaks following WPV importations into previously polio-free countries remain an ongoing risk until polio is eradicated. The GPEI Strategic Plan for 2010-2012 set the following two goals for outbreak control: 1) end outbreaks occurring in 2009 by mid-2010 and 2) end outbreaks occurring during 2010 to mid-2012 within 6 months of confirmation. This report describes new outbreaks that have occurred in the World Health Organization (WHO) European Region and updates previous reports on the status of outbreaks in Africa and Asia. In 2010, the first WPV importation into the European Region since the region was declared polio-free in 2002 resulted in 476 confirmed cases: 458 in Tajikistan, 14 in Russia, three in Turkmenistan, and one in Kazakhstan. In Africa and Asia, 11 new importations into six countries were observed in 2010; 30 WPV importations that occurred during 2008-2009 resulted in 215 cases in 15 African countries during 2009-2010. An outbreak is considered interrupted if 6 months have elapsed since the latest confirmed case and surveillance performance indicators meet WHO standards. All 2009 outbreaks in Africa appear to have been interrupted, and 2010 outbreaks in three countries appear to have been interrupted. Maintaining high routine vaccination coverage and sensitive surveillance at all times and rapidly instituting additional immunization programs to control outbreaks are key to limiting and stopping the spread of WPV.


Detection of human coronavirus NL63 and OC43 in children with acute respiratory infections in Niigata, Japan, between 2010 and 2011.


1802: DISSEMINATED BLASTOMYCOSIS PRESENTING WITH ACUTE NEUROLOGIC SYMPTOMS DUE TO CEREBELLAR ABSCESSES.


title: Evolutionary insights into the ecology of coronaviruses.

text: Although many novel members of the Coronaviridae have recently been recognized in different species, the ecology of coronaviruses has not been established. Our study indicates that bats harbor a much wider diversity of coronaviruses than any other animal species. Dating of different coronavirus lineages suggests that bat coronaviruses are older than those recognized in other animals and that the human severe acute respiratory syndrome (SARS) coronavirus was directly derived from viruses from wild animals in wet markets of southern China. Furthermore, the most closely related bat and SARS coronaviruses diverged in 1986, an estimated divergence time of 17 years prior to the outbreak, suggesting that there may have been transmission via an unknown intermediate host. Analysis of lineage-specific selection pressure also indicated that only SARS coronaviruses in civets and humans were under significant positive selection, also demonstrating a recent interspecies transmission. Analysis of population dynamics revealed that coronavirus populations in bats have constant population growth, while viruses from all other hosts show epidemic-like increases in population. These results indicate that diverse coronaviruses are endemic in different bat species, with repeated introductions to other animals and occasional establishment in other species. Our findings suggest that bats are likely the natural hosts for all presently known coronavirus lineages and that all coronaviruses recognized in other species were derived from viruses residing in bats. Further surveillance of bat and other animal populations is needed to fully describe the ecology and evolution of this virus family.


title: Nebulized versus Standard Local Application of Lidocaine during Flexible Bronchoscopy: A Randomized Controlled Trial.

text: BACKGROUND Endobronchial administration of local anesthetics such as lidocaine is often used for cough suppression during bronchoscopy. To achieve a better distribution of lidocaine in the tracheobronchial tree, spray catheters have been developed, allowing nebulization of the local anesthetic solution. However, there are little data on the efficacy and safety of this approach, or on the consumption of sedative drugs and lidocaine during nebulized administration. OBJECTIVES To investigate the tolerability of nebulized lidocaine compared to conventional lidocaine administration via syringe through the working channel of the bronchoscope in patients undergoing bronchoscopy. Consumption of sedative drugs and lidocaine was also compared between the two lidocaine administration approaches. METHODS Patients requiring bronchoscopy with endobronchial or transbronchial biopsy were randomly assigned to receive topical lidocaine either via syringe or via nebulizer. Endpoints were consumption of lidocaine and sedative drugs, as well as patient tolerance and safety. RESULTS Thirty patients were included, 15 in each group. Patients in the nebulizer group required lower doses of endobronchial lidocaine (184.7 ± 67.98 vs. 250.7 ± 21.65 mg, p = 0.0045) and intravenous fentanyl (0.033 ± 0.041 vs. 0.067 ± 0.045 mg, p = 0.0236) than those in the syringe group; midazolam or propofol dosages did not differ between the two groups. In addition, there were no between-group differences in patient tolerance or safety (all p > 0.05). CONCLUSION Endobronchial administration of lidocaine during bronchoscopy via nebulizer was found to be well tolerated and safe and was associated with reduced lidocaine and fentanyl dosages compared to administration via syringe.


title: Virus-specific and bystander CD8 T cells recruited during virus-induced encephalomyelitis.

text: Neurotropic coronavirus-induced encephalitis was used to evaluate recruitment, functional activation, and retention of peripheral bystander memory CD8+ T cells. Mice were first infected with recombinant vaccinia virus expressing a non-cross-reactive human immunodeficiency virus (HIV) epitope, designated p18. Following establishment of an endogenous p18-specific memory CD8+ T-cell population, mice were challenged with coronavirus to directly compare recruitment, longevity, and activation characteristics of both primary coronavirus-specific and bystander memory populations trafficking into the central nervous system (CNS). HIV-specific memory CD8+ T cells were recruited early into the CNS as components of the innate immune response, preceding CD8+ T cells specific for the dominant coronavirus epitope, designated pN. Although pN-specific T-cell numbers gradually exceeded bystander p18-specific CD8+ T-cell numbers, both populations peaked concurrently within the CNS. Nevertheless, coronavirus-specific CD8+ T cells were preferentially retained. By contrast, bystander CD8+ T-cell numbers declined to background numbers following control of CNS virus replication. Furthermore, in contrast to highly activated pN-specific CD8+ T cells, bystander p18-specific CD8+ T cells recruited to the site of inflammation maintained a nonactivated memory phenotype and did not express ex vivo cytolytic activity. Therefore, analysis of host CD8+ T-cell responses to unrelated infections demonstrates that bystander memory CD8+ T cells can comprise a significant proportion of CNS inflammatory cells during virus-induced encephalitis. However, transient CNS retention and the absence of activation suggest that memory bystander CD8+ T cells may not overtly contribute to pathology in the absence of antigen recognition.


title: Apical budding of a recombinant influenza A virus expressing a hemagglutinin protein with a basolateral localization signal.

text: Influenza virions bud preferentially from the apical plasma membrane of infected epithelial cells, by enveloping viral nucleocapsids located in the cytosol with its viral integral membrane proteins, i.e., hemagglutinin (HA), neuraminidase (NA), and M2 proteins, located at the plasma membrane. Because individually expressed HA, NA, and M2 proteins are targeted to the apical surface of the cell, guided by apical sorting signals in their transmembrane or cytoplasmic domains, it has been proposed that the polarized budding of influenza virions depends on the interaction of nucleocapsids and matrix proteins with the cytoplasmic domains of HA, NA, and/or M2 proteins. Since HA is the major protein component of the viral envelope, its polarized surface delivery may be a major force that drives polarized viral budding. We investigated this hypothesis by infecting MDCK cells with a transfectant influenza virus carrying a mutant form of HA (C560Y) with a basolateral sorting signal in its cytoplasmic domain. C560Y HA was expressed nonpolarly on the surface of infected MDCK cells. Interestingly, viral budding remained apical in C560Y virus-infected cells, and so did the location of NP and M1 proteins at late times of infection. These results are consistent with a model in which apical viral budding is a shared function of various viral components rather than a role of the major viral envelope glycoprotein HA.


title: Replication of murine coronaviruses in mouse embryonic stem cell lines in vitro.

text: Replication of murine coronaviruses in eight mouse embryonic stem (ES) cell lines of several genetic backgrounds was examined. Both mouse hepatitis virus (MHV) type 2 and MHV, strain A59 replicated well with no or minimal cytopathic effect in all the ES cell lines tested. The results suggest the possibility that MHV-infected ES cells may disseminate MHV in mouse colonies due to embryo manipulation.


Acute immunodeficiency, multiple organ injury, and the pathogenesis of SARS.


title: Nucleocapsid or spike protein-specific CD4+ T lymphocytes protect against coronavirus-induced encephalomyelitis in the absence of CD8+ T cells.

text: To investigate the antiviral CD4+ T cell response in coronavirus MHV-JHM-induced encephalomyelitis, spleen and thymic lymphocytes from diseased rats were stimulated in culture with virus Ag, expanded and tested for their specificity to viral proteins and nucleocapsid (N) and spike (S) proteins that had been expressed in bacteria. A strong T cell response specific for N was measurable during acute disease, whereas S-specific T cells were only detectable in rats with a later onset of disease. CD4+ T cell lines with specificity for virus and either N or S protein were established and their influence on the course of a mouse hepatitis virus-JHM infection was investigated. All lines were of the CD4+ phenotype. Both N and S protein-specific CD4+ T cells conferred protection to infected Lewis rats and reduced the amount of infectious virus in the central nervous system. After transfer of CD4+ T cells and challenge with virus, an increase in the antiviral IgM response occurred, but neutralizing antibodies were not detectable during the period of virus clearance. Previous CD8+ cell depletion did not abrogate protection mediated by CD4+ T cell line transfer.


title: The biophysics of renal sympathetic denervation using radiofrequency energy.

text: Renal sympathetic denervation is currently performed in the treatment of resistant hypertension by interventionists who otherwise do not typically use radiofrequency (RF) energy ablation in their clinical practice. Adequate RF lesion formation is dependent upon good electrode-tissue contact, power delivery, electrode-tissue interface temperature, target-tissue impedance and the size of the catheter's active electrode. There is significant interplay between these variables and hence an appreciation of the biophysical determinants of RF lesion formation is required to provide effective and safe clinical care to our patients. In this review article, we summarize the biophysics of RF ablation and explain why and how complications of renal sympathetic denervation may occur and discuss methods to minimise them.


title: Dynamic transmission modeling: a report of the ISPOR-SMDM Modeling Good Research Practices Task Force Working Group-5.

text: The transmissible nature of communicable diseases is what sets them apart from other diseases modeled by health economists. The probability of a susceptible individual becoming infected at any one point in time (the force of infection) is related to the number of infectious individuals in the population, will change over time, and will feed back into the future force of infection. These nonlinear interactions produce transmission dynamics that require specific consideration when modeling an intervention that has an impact on the transmission of a pathogen. Best practices for designing and building these models are set out in this paper.


title: [Analysis of relation between the usage of corticosteroid in treatment and arthralgia as a sequela of SARS patients].

text: OBJECTIVE To analyze relation between the use of corticosteroids in the treatment for severe acute respiratory syndrome (SARS) patients and arthralgia as a sequela. METHODS Clinical date of 30 SARS patients without other diseases in whom corticosteroid was used were reviewed including total dosage, duration of use, the highest dosage and its duration, and speed of reduction in dosage. The information about arthralgia was investigated one month after discharge of SARS patients from the hospital. RESULTS The average total dosage of methylprednisolone was (4 244.16+/-2 292.30) mg, and the duration of use of the treatment was (25.36+/-5.88) days (ranging from 12 to 35 days). The maximum dosage was (321.33+/-174.03) mg/d, and the duration of its use was (7.73+/-4.08) days. The speed of reduction of dosage of corticosteroids was (21.33+/-10.18) mg/d. There were 26 of 30 patients (86.67 percent) experienced arthralgia symptom during convalescence. In 3.6 percent of patients arthralgia occurred within one month after SARS, 53.85 percent of the patients experienced low-grade arthralgia. By unifactor analysis, the total dosage and its duration of use, the highest dosage and its duration, speed of reduction of dosage of corticosteroids were correlated with the degree of arthralgia, respectively. The duration of arthralgia was correlated with the total dosage, the duration of high dosage, and high dosage. Age was not correlated with either the degree or the persisting time of arthralgia. The degree of arthralgia was only correlated with the total dosage, and the duration of arthralgia was correlated with administration time of glucocorticosteroids by multifactor analysis. CONCLUSION There is a dosage- effect relation between the degree of arthralgia and the total dosage of corticosteroid, and a time-effect relation between the duration of arthralgia and length of the use of corticosteroids.


New steam from an old cauldron--the physician-supply debate.


title: High-frequency versus low-frequency microwave ablation in malignant liver tumours: evaluation of local tumour control and survival.

text: PURPOSE To compare local tumour control and survival rates in patients with liver metastases treated with microwave ablation (MWA), using either a low-frequency (LF) (915 MHz) or high-frequency (HF) system (2.45 GHz). MATERIALS AND METHODS The retrospective study included 221 patients (mean age: 61.7 years) with 356 malignant hepatic lesions. Ninety-four patients with 133 lesions underwent LF-MWA between September 2008 and February 2011, while 127 patients with 223 lesions were treated with HF-MWA between March 2011 and July 2013. MRI was performed after 24 h from each procedure and at 3, 6, 9, 12, 18 and 24 months post-ablation. Both groups were compared with the Fisher's exact test. Survival rates were calculated using the Kaplan-Meier test. RESULTS The mean initial ablation volume of LF-MWA was nearly half of HF-MWA (19.1 mL vs. 39.9 mL). The difference in volume between both systems was significant (p < .0001). With LF-MWA, 39/133 lesions (29.32%) progressed at follow-up while the number of lesions which progressed with HF-MWA was 10/223 (4.5%). The mean time to progression was 5.03 and 5.31 months for the lesions treated with LF-MWA and HF-MWA, respectively. The difference between both systems was significant (p = .00059). The 1-, 2- and 4-year overall survival rates for curative indication were 98.9%, 95.7% and 82.9% for LF-MWA, respectively, and were 100%, 97.6% and 92.9% for HF-MWA, respectively. The difference in survival rates was not significant (p > .05). CONCLUSION Both LF- and HF-MWA systems are effective treatment options for oligonodular liver malignant lesions, but significantly higher ablation volumes, longer time to progression and lower progression rates were observed in HF-MWA.


title: Membrane-bound carboxypeptidase E facilitates the entry of eosinophil cationic protein into neuroendocrine cells.

text: ECP (eosinophil cationic protein) is a major component of eosinophil granule proteins, and is used as a clinical biomarker for asthma and allergic inflammatory disease. ECP has been implicated in damage to the cell membrane of many tissue types, but the mechanism is not well known. In the present study, mECP-eGFP-6H, a recombinant fusion protein containing mature ECP (mECP), enhanced green fluorescence protein (eGFP) and a His(6) tag (6H), has been expressed, purified and added to GH3 neuroendocrine cells to study the internalization ability of ECP. We found that mECP-eGFP-6H entered into GH3 neuroendocrine cells and inhibited the growth of the cells with an IC(50) of 0.8 microM. By yeast two-hybrid screening and immunoprecipitation, we have identified a specific protein-protein interaction between mECP and CPE (carboxypeptidase E), a well characterized metalloprotease. Further in vivo yeast two-hybrid screening has also revealed that residues 318-387 located in a region of unknown function in mature CPE are indispensable for association with mECP. In addition, the uptake of mECP-eGFP-6H is suppressed by dominant-negative expression of the recycling defect mutant pre-pro-HA-CPE(S471A,E472A) in GH3 cells, suggesting that the entry of mECP-eGFP-6H is associated with the recycling of CPE in GH3 cells. Taken together, we have demonstrated that CPE possesses a novel function to facilitate the entry of ECP to neuroendocrine cells, and such an endocytotic process allows the cytotoxic ECP to inhibit growth of the target cells.


title: Stemming COVID-19 in Cuba: Strengths, Strategies, ChallengesFrancisco Durán MD.

text: Dr Durán is a native of eastern San-tiago de Cuba and his early medical career began in this mountainous re-gion, where he also headed provin-cial prevention and treatment of HIV/AIDS. He went on to become rector of the Medical University of Santiago de Cuba and provincial health direc-tor. Later in Havana, Dr Durán was director of medical education and vice minister at the Ministry of Public Health (MINSAP). Abroad, he served as advisor to Angola's Minister of Health, and on his return, as deputy director of Cuba's Pedro Kourí Tropical Medicine Institute (IPK). Dr Durán has been "battle-tested" over the years by his involvement in stemming dengue epidemics and other infectious dis-ease outbreaks, good preparation for his current position as National Direc-tor of Epidemiology. Today, his is the voice and the face on the 11:00 AM briefing carried daily by Cuban television, reporting the latest data on the un-folding COVID-19 pandemic globally, in the Americas and.


title: A serological survey of canine respiratory coronavirus and canine influenza virus in Korean dogs.

text: The relationship between canine respiratory coronavirus (CRCoV) and canine influenza virus (CIV) seropositivity in dogs in Korea was examined. Sixty-two of the 483 samples (12.8%) were seropositive for CRCoV by indirect fluorescent antibody (IFA) analysis. Nineteen animals were seropositive for CIV by ELISA out of the 385 samples tested. Serum antibodies for both viruses were detected in 6 of the 483 dogs sampled, suggesting that these viruses are present in dogs in Korea. Although the role of CRCoV in canine infectious tracheobronchitis has not been fully elucidated, co-infection with CIV may synergistically worsen respiratory clinical signs and result in more severe canine tracheobronchitis.


title: Characterization in vitro of an autocatalytic processing activity associated with the predicted 3C-like proteinase domain of the coronavirus avian infectious bronchitis virus.

text: A region of the infectious bronchitis virus (IBV) genome between nucleotide positions 8693 and 10927 which encodes the predicted 3C-like proteinase (3CLP) domain and several potential cleavage sites has been clones into a T7 transcription vector. In vitro translation of synthetic transcripts generated from this plasmid was not accompanied by detectable processing activity of the nascent polypeptide unless the translation was carried out in the presence of microsomal membrane preparations. The processed products so obtained closely resembled in size those expected from cleavage at predicted glutamine-serine (Q/S) dipeptides and included a protein with a size of 35 kDa (p35) that corresponds to the predicted size of 3CLP. Efficient processing was dependent on the presence of membranes during translation; processing was found to occur when microsomes were added posttranslationally, but only after extended periods of incubation. C-terminal deletion analysis of the encoded polyprotein fragment revealed that cleavage activity was dependent on the presence of most but not all of the downstream and adjacent hydrophobic region MP2. Dysfunctional mutagenesis of the putative active-site cysteine residue of 3CLP to either serine or alanine resulted in polypeptides that were impaired for processing, while mutagenesis at the predicted Q/S release sites implicated them in the release of the p35 protein. Processed products of the wild-type protein were active in trans cleavage assays, which were used to demonstrate that the IBV 3CLP is sensitive to inhibition by both serine and cysteine protease class-specific inhibitors. These data reveal the identity of the IBV 3C-like proteinase, which exhibits characteristics in common with the 3C proteinases of picornaviruses.


title: Rotavirus virus-like particles administered mucosally induce protective immunity.

text: We have evaluated the immunogenicity and protective efficacy of rotavirus subunit vaccines administered by mucosal routes. Virus-like particles (VLPs) produced by self-assembly of individual rotavirus structural proteins coexpressed by baculovirus recombinants in insect cells were the subunit vaccine tested. We first compared the immunogenicities and protective efficacies of VLPs containing VP2 and VP6 (2/6-VLPs) and G3 2/6/7-VLPs mixed with cholera toxin and administered by oral and intranasal routes in the adult mouse model of rotavirus infection. VLPs administered orally induced serum antibody and intestinal immunoglobulin A (IgA) and IgG. The highest oral dose (100 microg) of VLPs induced protection from rotavirus challenge (> or = 50% reduction in virus shedding) in 50% of the mice. VLPs administered intranasally induced higher serum and intestinal antibody responses than VLPs administered orally. All mice receiving VLPs intranasally were protected from challenge; no virus was shed after challenge. Since there was no difference in immunogenicity or protective efficacy between 2/6- and 2/6/7-VLPs, protection was achieved without inclusion of the neutralization antigens VP7 and VP4. We also tested the immunogenicities and protective efficacies of 2/6-VLPs administered intranasally without the addition of cholera toxin. 2/6-VLPs administered intranasally without cholera toxin induced lower serum and intestinal antibody titers than 2/6-VLPs administered with cholera toxin. The highest dose (100 microg) of 2/6-VLPs administered intranasally without cholera toxin resulted in a mean reduction in shedding of 38%. When cholera toxin was added, higher levels of protection were achieved with 10-fold less immunogen. VLPs administered mucosally offer a promising, safe, nonreplicating vaccine for rotavirus.


title: Enteric coronavirus infection in a juvenile dromedary (Camelus dromedarius).

text: A case of an enteric coronavirus infection in a 6-week-old dromedary calf is described. The animal had diarrhea for 5 days and died despite symptomatic treatment. Numerous viral particles, approximately 140 nm in diameter, with club-like projections were detected in the feces by electron microscopy. These characteristics were consistent with a coronavirus. Immunohistochemical reactivity with 2 antigenic group II coronavirus-specific antibodies confirmed the presence of viral antigen in colonic epithelial cells. The death of the animal was attributed to a neutrophilic and emphysematous colitis that likely was caused by an infection with a Clostridium sp.


title: Single-access transumbilical laparoscopic unroofing of a giant hepatic cyst using reusable instruments.

text: INTRODUCTION Single-access laparoscopy has garnered growing interest in recent years in an attempt to improve cosmesis, reduce postoperative pain, and minimize abdominal wall trauma. CASE DESCRIPTION A female patient suffering from a symptomatic giant biliary cyst of the liver segments 4-7-8 was admitted for transumbilical single-access laparoscopic cyst unroofing. The procedure was performed using a standard 11-mm reusable trocar for a 10-mm, 30 degree-angled, rigid scope and curved reusable instruments inserted transumbilically without trocars. Operative time was 90 minutes, and the final incision length was 14 mm. The use of minimal pain medication permitted discharge on the third postoperative day, and after 25 months, the patient remains asymptomatic with a no visible umbilical scar. CONCLUSIONS Giant biliary cysts can be removed by single-access laparoscopy. Because of this technique, surgeons work in ergonomic positions, and the cost of the procedure remains similar to that of the multitrocar technique. The incision length and the use of pain medication are kept minimal as well.


title: A case of imported Middle East Respiratory Syndrome coronavirus infection and public health response, Greece, April 2014.

text: On 18 April 2014, a case of Middle East Respiratory Syndrome coronavirus (MERS-CoV) infection was laboratory confirmed in Athens, Greece in a patient returning from Jeddah, Saudi Arabia. Main symptoms upon initial presentation were protracted fever and diarrhoea, during hospitalisation he developed bilateral pneumonia and his condition worsened. During 14 days prior to onset of illness, he had extensive contact with the healthcare environment in Jeddah. Contact tracing revealed 73 contacts, no secondary cases had occurred by 22 April.


title: Role of pH in syncytium induction and genome uncoating of avian infectious bronchitis coronavirus (IBV).

text: Syncytium formation induced in Vero cells by IBV-Beaudette was optimal at pH 6.7 and absent at pH 7.5. Reduction of IBV-Beaudette replication by ammonium chloride, which raises the pH in endosomes, further indicated that uncoating of the virus genome probably occurred within the acidic environment of endosomes. However, this would not appear to apply to all IBV strains, since one was unaffected by the drug. It would appear that some strains of IBV have the capacity to fuse with the plasma membrane at neutral pH, thereby leading to genome uncoating at that site. Other strains of IBV might also uncoat at the plasma membrane when the extracellular environment is slightly less than pH7.


title: [Pharmacotherapy of severe acute respiratory syndrome (SARS)].

text: Severe acute respiratory syndrome (SARS) constitutes the first new infectious disease of the current millennium. It is caused by the novel SARS-Coronavirus (SARS-CoV). SARS is related to a high morbidity and mortality and first appeared during an epidemic in 2002 - 2003. To date no specific therapy against the SARS-CoV is available. Due to the rapid spread of SARS during the epidemics in 2002 - 2003, randomised and controlled multicentre studies were not performed. Therefore, general guidelines have not been developed. Since the outbreak, scientists have been testing potential antiviral substances using in vitro and animal models. This study analyses the presently available in vitro and in vivo data on the pharmacotherapy of SARS.


title: SARS assessment clinic: a rapid response to an infectious outbreak.

text: The Toronto SARS outbreak began in February 2003 and lasted more than 16 weeks. The city and its health care system faced enormous challenges in responding to this new infectious disease, learning about its transmission, diagnosis and treatment, in containing its spread and in coping with its socioeconomic impact. As the site of a significant cluster of cases in the second wave of the outbreak, North York General Hospital (NYGH) quickly adapted many components of its operations, focusing on the fight against SARS. In order to assess potential SARS cases in a safe, efficient and effective manner, NYGH established a SARS assessment clinic. We describe the design features, construction, layout and operation of this clinic. This type of clinic can be rapidly deployed and may be of great value during future infectious outbreaks, including pandemic influenza.


title: Ecological and anthropogenic drivers of rabies exposure in vampire bats: implications for transmission and control.

text: Despite extensive culling of common vampire bats in Latin America, lethal human rabies outbreaks transmitted by this species are increasingly recognized, and livestock rabies occurs with striking frequency. To identify the individual and population-level factors driving rabies virus (RV) transmission in vampire bats, we conducted a longitudinal capture-recapture study in 20 vampire bat colonies spanning four regions of Peru. Serology demonstrated the circulation of RV in vampire bats from all regions in all years. Seroprevalence ranged from 3 to 28 per cent and was highest in juvenile and sub-adult bats. RV exposure was independent of bat colony size, consistent with an absence of population density thresholds for viral invasion and extinction. Culling campaigns implemented during our study failed to reduce seroprevalence and were perhaps counterproductive for disease control owing to the targeted removal of adults, but potentially greater importance of juvenile and sub-adult bats for transmission. These findings provide new insights into the mechanisms of RV maintenance in vampire bats and highlight the need for ecologically informed approaches to rabies prevention in Latin America.


title: The peptidases from fungi and viruses.

text: Fungi and viruses encode a variety of peptidases having a plethora of functions. Many fungal peptidases are extracellular and are likely used to degrade proteins in their environment. Viral peptidases are processing enzymes, intimately involved in the virus infectious cycle. The viral RNA genome is translated by the host-cell machinery into a large polyprotein that is cleaved by the viral peptidases into mature capsid proteins, non-structural proteins and enzymes. I review the structure and catalytic mechanism of scytalidoglutamic peptidase isolated from the wood-destroying fungus Scytalidium lignicolum. This enzyme has a unique beta-sandwich fold and a novel catalytic mechanism based on a glutamate, a glutamine and a nucleophilic water molecule. Hepatitis A virus (HAV) 3C peptidase was the first structure identified for a viral 3C enzyme that exhibited the three-dimensional fold of the chymotrypsin family of serine peptidases but had a cysteine sulfur atom instead of the serine oxygen as the nucleophile. The structure of HAV 3C was unusual in that the Asp residue expected as the third member of the catalytic triad did not interact with the general base His. The present structure is of a beta-lactone-inhibited version of HAV 3C that has a restored catalytic triad.


title: Reactivities of 4 murine coronavirus antigens with immunized or naturally infected rat sera by enzyme linked immunosorbent assay.

text: Four murine coronavirus antigens, sialodacryoadenitis virus (SDAV) strain TG, Parker's rat coronavirus (PCV) strain 8190, mouse hepatitis virus (MHV) strains S and NuU, were examined for their reactivities to hyperimmunized and naturally infected rat sera by ELISA. With the immunized sera, SDAV and PCV antigens reacted best with respective homologous sera. MHV antigens reacted with all antisera, anti-SDAV, anti-PCV, and anti-MHV-S at approximately the same level, and MHV-S showed a slightly higher reactivity than MHV-NuU. The reactivities of the sera from various colonies to these antigens were in the order--from high to low--of SDAV, MHV-S, MHV-NuU, and PCV. None of sera negative for SDAV antigen reacted positively to the other antigens. Within the sera positive for SDAV, the positivities were in the order of MHV-S, MHV-NuU, and PCV. These results suggested that, although homologous antigens are best to detect SDAV or PCV infection by ELISA, MHV antigen can be used if highly cross-reactive viral strain is selected.


title: Coronavirus infection in the laboratory rat: immunization trials using attenuated virus replicated in L-2 cells.

text: Sixty-nine specific pathogen-free male Wistar rats approximately eight weeks of age were used to evaluate the efficacy of an attentuated strain of sialodacryoadenitis (SDA) virus in providing protection against infection on subsequent challenge with virulent SDA virus. Fifty-four animals were inoculated intranasally with approximately 10(3.5) median cell culture infectious doses of the 25th passage of SDA virus in L-2 cells. Randomly-selected vaccinated animals were killed in order to evaluate the safety and efficacy of attenuated virus by histopathological examination of the salivary glands, lacrimal glands, and lower respiratory tract, and titration of sera for antibody to SDA virus. At three months and six months postvaccination (pv), animals were selected at random and challenged with virulent SDA virus. Seronegative, age-matched animals were also challenged, and served as controls. In animals examined at six to ten days pv, lesions were absent in submandibular and parotid salivary glands and lacrimal glands, but transient lesions were present in major airways of the lower respiratory tract. In a comparison of the incidence and extent of lesions, and antibody titers in challenged vaccinates and seronegative controls, lesions were minimal or absent in vaccinates compared to challenged naive rats, particularly in animals inoculated at three months pv. In addition, antibody titers in challenged vaccinates were much higher than were postinoculation titers in inoculated controls. In a comparison of lesions in salivary and lacrimal glands in vaccinated and control animals challenged at six months pv, there was a significant reduction in the number of animals without lesions in the vaccinated group (p = less than 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)


title: Array-based binary analysis for bacterial typing.

text: An allele-specific oligonucleotide microarray was developed for rapid typing of pathogens based on analysis of genomic variations. Using a panel of Escherichia coli strains as a model system, selected loci were sequenced to uncover differences, such as single- or multiple-nucleotide polymorphisms as well as insertion/deletions (indels). While typical genomic profiling experiments employ specific sequences targeted to genomic DNA unique to a single strain or virulent gene, the present array is designed to type bacteria based on a patterned signature response across multiple loci. In the signature concept, all strains are interrogated by hybridizing their amplified DNA to an array containing multiple probe sequences. Allele-specific oligonucleotide probe sequences targeting each of these variable regions were synthesized and included in a custom fiber-optic array. For each locus, a set of specific probe sequences is selected, such that hybridization gives a binary signal/no signal response to each of the probes. Using this strategy for multiple loci, many pathogens or microorganisms could be classified using a limited number of probes. Because of the advantages of the fiber-optic array platform over other array formats, including sensitivity and speed, the platform described in this paper is capable of supporting a high-throughput diagnostic strategy.


title: Progress toward an HIV vaccine.

text: The development of an HIV vaccine is proving to be an unprecedented challenge. The difficulty in creating this vaccine arises from the enormous genetic variation of the virus and the unusual importance of cytotoxic T lymphocytes (CTL) in controlling its spread. Whereas traditional vaccine strategies are unlikely to confer safe and effective HIV protection, novel strategies for eliciting CTL have provided substantial clinical benefits in nonhuman primate model systems. These vaccine strategies, including plasmid DNA and live recombinant vectors, are currently being evaluated in human clinical trials.


title: Nationwide survey of antibodies to bovine coronavirus in bulk milk from Swedish dairy herds.

text: Bulk milk samples from 2236 dairy herds randomly selected throughout Sweden in proportion to region and herd size were analysed for antibodies to bovine coronavirus (BCV) in an ELISA. The results were expressed as optical density (OD) values and an OD > 0.04 was considered positive. Eighty-nine per cent of the samples were antibody-positive and 52 per cent had high levels of antibodies to BCV (an OD > 0.70). There were significantly higher OD values (P < 0.001) and fewer antibody-negative samples (P < 0.001) from larger herds than from smaller herds. There were also significantly higher OD values and fewer antibody-negative samples from herds in southern Sweden than from herds in northern Sweden (P < 0.001 and P < 0.001, respectively). These results indicate a higher frequency of BCV infections in larger herds and in herds in southern Sweden.


title: Spatio-temporal and stochastic modelling of severe acute respiratory syndrome.

text: This study describes the development of a spatio-temporal disease model based on the episodes of severe acute respiratory syndrome (SARS) that took place in Hong Kong in 2003. In contrast to conventional, deterministic modelling approaches, the model described here is predominantly spatial. It incorporates stochastic processing of environmental and social variables that interact in space and time to affect the patterns of disease transmission in a community. The model was validated through a comparative assessment between actual and modelled distribution of diseased locations. Our study shows that the inclusion of location-specific characteristics satisfactorily replicates the spatial dynamics of an infectious disease. The Pearson's correlation coefficients for five trials based on 3-day aggregation of disease counts for 1-3, 4-6 and 7-9 day forecasts were 0.57- 0.95, 0.54-0.86 and 0.57-0.82, respectively, while the correlation based on 5-day aggregation for the 1-5 day forecast was 0.55- 0.94 and 0.58-0.81 for the 6-10 day forecast. The significant and strong relationship between actual results and forecast is encouraging for the potential development of an early warning system for detecting this type of disease outbreaks.


title: [The clinical characteristics and outcome of 45 early stage patients with SARS].

text: OBJECTIVE To investigate the clinical characteristics and therapeutic effect in the early stage of patients with Severe Acute Respiratory Syndrome (SARS). METHODS A total of 45 clinical diagnosed SARS patients were enrolled in the study. RESULTS 43 (95.6%) of all 45 patients had exposure to SARS patients and the average incubation time was (7.3 +/- 4.7) days (range 2 to 20 days). The clinical features including fatigue (60%), dry cough (68.9%), dyspnea (53.3%), joint pain (26.7%) and diarrhea (26.7%). The laboratory test show that 33% patients with leukopenia and thrombocytopenia, 53.3% with elevated serum aminotransferase and 26.6% with increased creatinine kinase levels. The occurrence of abnormal chest radiographs was (4.2 +/- 2.9) days from the onset, and CT scanning was better than X-ray in the early diagnosis. The average time for the progression and improvement of the chest radiographs were (9.3 +/- 4.1) and (14.5 +/- 5.0) days, respectively. The administration of corticosteroids was started (4.8 +/- 3.5) days from the onset and the maximal dosage using of steroids was (8.4 +/- 3.9) days. There was a significant difference in the dosage of steroid between severe and mild patients [(610.6 +/- 212.4) mg/day vs (268.1 +/- 202.5) mg/day, P < 0.001)]. The IgG antibody was detected in 22 out of 34 (64.7%) patients and the average time for the occurrence of antibody was (18.2 +/- 6.4) days (range 9-27 days). CONCLUSION Exposure to the SARS patients happened in most patients. The most common clinical features were fever and fatigue. The most common laboratory abnormalities were leukopenia and thrombocytopenia. Serum antibody was not detected until on the 9th day of the course of illness; CT is a more sensitive method for detection of the abnormal chest radiographs than regular X-ray test. The early usage of cortiscosteroids was an efficient method for the treatment, and the dosage of steroids should be individualized.


title: The polypeptide of Mr 14,000 of porcine transmissible gastroenteritis virus: gene assignment and intracellular location.

text: Synthetic oligopeptides, corresponding to an amino acid sequence encoded by a potential Mr 9000 product's open reading frame (ORF-4) at the 3' terminus of the transmissible gastroenteritis virus genome, were used to generate rabbit antiserum. These antibodies produced immune complexes with an Mr 14,000 (14K) polypeptide in infected cells. The 14K product was shown by immune fluorescence to become associated with the cell nucleus, correlating with the onset of nuclear vacuolation, and suggesting a role in pathogenesis for the ORF-4 gene.


title: Efficacy of disinfectants and hand sanitizers against avian respiratory viruses.

text: Disinfectants play a major role in the control of animal diseases by decontaminating the farm environment. We evaluated the virucidal efficacy of nine commonly used disinfectants on a nonporous surface contaminated experimentally with avian metapneumovirus (aMPV), avian influenza virus, or Newcastle disease virus (NDV). Phenolic compounds and glutaraldehyde were found to be the most effective against all three viruses. Quaternary ammonium compounds were effective against aMPV but not against the other two viruses. In addition, efficacy of commercially available hand sanitizers was evaluated on human fingers contaminated with aMPV and NDV. All three hand sanitizers tested were found to be effective against both viruses within 1 min of application on fingers.


title: Subcellular localization and targeting of N-acetylglucosaminyl phosphatidylinositol de-N-acetylase, the second enzyme in the glycosylphosphatidylinositol biosynthetic pathway.

text: The second step in glycosylphosphatidylinositol biosynthesis is the de-N-acetylation of N-acetylglucosaminylphosphatidylinositol (GlcNAc-PI) catalyzed by N-acetylglucosaminylphosphatidylinositol deacetylase (PIG-L). Previous studies of mouse thymoma cells showed that GlcNAc-PI de-N-acetylase activity is localized to the endoplasmic reticulum (ER) but enriched in a mitochondria-associated ER membrane (MAM) domain. Because PIG-L has no readily identifiable ER sorting determinants, we were interested in learning how PIG-L is localized to the ER and possibly enriched in MAM. We used HeLa cells transiently or stably expressing epitope-tagged PIG-L variants or chimeric constructs composed of elements of PIG-L fused to Tac antigen, a cell surface protein. We first analyzed the subcellular distribution of PIG-L and Glc-NAc-PI-de-N-acetylase activity and then studied the localization of Tac-PIG-L chimeras to identify sequence elements in PIG-L responsible for its subcellular localization. We show that human PIG-L is a type I membrane protein with a large cytoplasmic domain and that, unlike the result with mouse thymoma cells, both PIG-L and GlcNAc-PI-de-N-acetylase activity are uniformly distributed between ER and MAM in HeLa cells. Analyses of a series of Tac-PIG-L chimeras indicated that PIG-L contains two ER localization signals, an independent retention signal located between residues 60 and 88 of its cytoplasmic domain and another weak signal in the luminal and transmembrane domains that functions autonomously in the presence of membrane proximal residues of the cytoplasmic domain that themselves lack any retention information. We conclude that PIG-L, like a number of other ER membrane proteins, is retained in the ER through a multi-component localization signal rather than a discrete sorting motif.


title: High-level expression and purification of human epidermal growth factor with SUMO fusion in Escherichia coli.

text: Human epidermal growth factor (hEGF) can stimulate the division of various cell types and has potential clinical applications. However, the high expression of active hEGF in Escherichia coli has not been successful, as the protein contains three intra-molecular disulfide bonds that are difficult to form correctly in the bacterial intracellular environment. To solve this problem, we fused the hEGF gene with a small ubiquitin-related modifier gene (SUMO) by synthesizing an artificial SUMO-hEGF fusion gene that was highly expressed in Origami (DE3) strain. The optimal expression level of the soluble fusion protein, SUMO-hEGF, was up to 38.9% of the total cellular protein. The fusion protein was purified by Ni-NTA affinity chromatography and cleaved by a SUMO-specific protease to obtain the native hEGF, which was further purified by Ni-NTA affinity chromatography. The result of the reverse-phase HPLC showed that the purity of the recombinant cleaved hEGF was greater than 98%. The primary structure of the purified hEGF was confirmed by N-terminal amino acid sequencing and MALDI-TOF mass spectroscopy analysis. Using the method of methylthiazoletetrazolium, the mitogenic activity on Balb/c 3T3 cells of the purified hEGF was comparable to that of commercial hEGF.


title: Protection and disinfection policies against SARS-CoV-2 (COVID-19).

text: In late December 2019, reports from China of the incidence of pneumonia with unknown etiology were sent to the World Health Organization (WHO). Shortly afterwards, the cause of this disease was identified as the novel beta-coronavirus, SARS-CoV-2, and its genetic sequence was published on January 12, 2020. Human-to-human transmission via respiratory droplets and contact with aerosol infected surfaces are the major ways of transmitting this virus. Here we attempted to collect information on virus stability in the air and on surfaces and ways of preventing of SARS-CoV-2 spreading.


title: Proteolytic processing of the coronavirus infectious bronchitis virus 1a polyprotein: identification of a 10-kilodalton polypeptide and determination of its cleavage sites.

text: Proteolytic processing of the polyprotein encoded by mRNA 1 is an essential step in coronavirus RNA replication and gene expression. We have previously reported that an open reading frame (ORF) 1a-specific proteinase of the picornavirus 3C proteinase group is involved in processing of the coronavirus infectious bronchitis virus (IBV) 1a/1b polyprotein, leading to the formation of a mature viral protein of 100 kDa. We report here the identification of a novel 10-kDa polypeptide and the involvement of the 3C-like proteinase in processing of the ORF 1a polyprotein to produce the 10-kDa protein species. By using a region-specific antiserum, V47, raised against a bacterial-viral fusion protein containing IBV sequence encoded between nucleotides 11488 and 12600, the 10-kDa polypeptide was detected in lysates from both IBV-infected and plasmid DNA-transfected Vero cells. Coexpression, deletion, and mutagenesis studies showed that this novel polypeptide was encoded by ORF 1a from nucleotide 11545 to 11878 and was cleaved from the 1a polyprotein by the 3C-like proteinase domain. Evidence presented suggested that a previously predicted Q-S (Q3783 S3784) dipeptide bond encoded by ORF 1a between nucleotides 11875 and 11880 was responsible for the release of the C terminus of the 10-kDa polypeptide and that a novel Q-N (Q3672 N3673) dipeptide bond encoded between nucleotides 11542 and 11547 was responsible for the release of the N terminus of the 10-kDa polypeptide.


title: Studies on the development of an ELISA kit for microbiological monitoring. 2. Improvement of the prototype ELISA kit with special references to mouse hepatitis virus antigen.

text: Improvement of the mouse hepatitis virus (MHV) antigen in a prototype ELISA kit was performed. Equivalent divalent antigens of MHV Nu-67 and S strains with a protein concentration of 10 micrograms/ml showed the best sensitivity and specificity for the detection of MHV and sialodacryoadenitis/Parker's rat coronavirus antibodies in mice and rats, respectively. An increase in the reliability of macroscopic evaluation of both antibody tests is expected by using the newly manufactured kit with the improved antigen.


title: Production systems for recombinant antibodies.

text: Recombinant antibodies are the fastest growing class of therapeutic proteins. Furthermore, antibodies are key detection reagents in research and diagnostics. The increasing demand for antibodies with regards to amount and quality resulted in the development of a variety of recombinant production systems employing gram-negative and gram-positive bacteria, yeast and filamentous fungi, insect cell lines as well as mammalian cell lines. More recently, antibodies were also successfully produced in transgenic plants and animals. Currently, the production of recombinant antibodies for therapy is performed in mammalian cell lines to reduce the risk of immunogenicity caused by non-human post-translational modifications, in particular glycosylation. However, novel strategies already allow human-like glycosylation patterns in yeast, insect cell lines and transgenic plants. Furthermore, therapeutic strategies not requiring glycosylation of the Fc portion have been conceived, most prominently using bispecific antibodies or scFv fusion proteins, which can be produced in bacteria. Here, we review all current antibody production systems considering their advantages and limitations with respect to intended applications.


title: trans-Complementation of flavivirus RNA polymerase gene NS5 by using Kunjin virus replicon-expressing BHK cells.

text: A BHK cell line persistently expressing a Kunjin (KUN) virus replicon RNA (repBHK, similar to our recently described ME/76Neo BHK cell line [A. A. Khromykh and E. G. Westaway, J. Virol. 71:1497-1505, 1997]) was used for rescue and propagation of KUN viruses defective in the RNA polymerase gene (NS5). A new infectious full-length KUN virus cDNA clone, FLSDX, prepared from our previously described cDNA clone pAKUN (A. A. Khromykh and E. G. Westaway, J. Virol. 68:4580-4588, 1994) and possessing approximately 10(5)-fold higher specific infectivity than that of pAKUN, was used for preparation of defective mutants. Deletions of the predicted RNA polymerase motif GDD (producing FLdGDD) and of one of the predicted methyltransferase motifs (S-adenosylmethionine [SAM] binding site, producing FLdSAM) were introduced separately into FLSDX. Transcription and transfection of FLdGDD and FLdSAM RNAs into repBHK cells but not into normal BHK cells resulted in their replication and the recovery of defective viruses able to replicate only in repBHK cells. Reverse transcription-PCR and sequencing analyses showed retention of the introduced deletions in the genomes of the recovered viruses. Retention of these deletions, as well as our inability to recover viruses able to replicate in normal BHK cells after prolonged incubation (for 7 days) of FLdGDD- or FLdSAM-transfected repBHK cells, excluded the possibility that recombination had occurred between the deleted defective NS5 genes present in transfected RNAs and the functional NS5 gene present in the repBHK cells. An RNA with a point mutation in the GDD motif (FLGVD) was also complemented in transfected repBHK cells, and defective virus was recovered by day 3 after transfection. However, in contrast to the results with FLdGDD and FLdSAM RNAs, prolonged (4 days or more) incubation of FLGVD RNA in normal BHK cells allowed recovery of a virus in which the GVD mutation had reverted via a single base change to the wild-type GDD sequence. Overall, these results represent the first demonstration of trans-complementation of defective flavivirus RNAs with deleterious deletions in the flavivirus RNA polymerase gene NS5. The complementation system described here may prove to be useful for the in vivo complementation of deletions and mutations affecting functional domains or the essential secondary structure in any of the other flavivirus nonstructural proteins.


Bone Infections, Sinusitis, Antihypertensive Therapy and COVID-19, Toilet Training, Amenorrhea.


title: Coronavirus-induced demyelination occurs in the absence of inducible nitric oxide synthase.

text: Demyelination induced by mouse hepatitis virus (MHV), strain JHM, is in large part immune mediated, but little is known about the mechanisms involved in this process. Previous results suggest that inducible nitric oxide synthase (NOS2) contributes transiently to MHV-induced demyelination. Herein, we show that equivalent amounts of demyelination were evident at day 12 after MHV infection in mice genetically deficient in NOS2 (NOS2(-/-)) and in C57BL/6 mice. Furthermore, using an established adoptive transfer model and pharmacological inhibitors of NOS2 function, we could demonstrate no effect on MHV-induced demyelination. These results indicate that NOS2 function is not required for demyelination in mice infected with MHV.


Isolation of hemagglutinating encephalomyelitis virus from respiratory tract of pigs in Japan.


Comparison of APTIMA COMBO 2 assay with polymerase chain reaction Roche assay for detection of Chlamydia trachomatis in endocervical swabs.


Middle East Respiratory Syndrome coronavirus - two years into the epidemic.


title: Prophylactic non-steroidal anti-inflammatory drugs for the prevention of macular oedema after cataract surgery.

text: BACKGROUND Macular oedema (MO) is the accumulation of extracellular fluid in the central retina (the macula). It may occur after cataract surgery and may give rise to poor visual outcome, with reduced visual acuity and distortion of the central vision. MO is often self-limiting with spontaneous resolution, but a small proportion of people with chronic persistent MO may be difficult to treat. Chronic oedema may lead to the formation of cystic spaces in the retina termed 'cystoid macular oedema' (CMO). Non-steroidal anti-inflammatory drugs (NSAIDs) are commonly used in cataract surgery and may reduce the chances of developing MO. OBJECTIVES The aim of this review is to answer the question: is there evidence to support the prophylactic use of topical NSAIDs either in addition to, or instead of, topical steroids postoperatively to reduce the incidence of macular oedema (MO) and associated visual morbidity. SEARCH METHODS We searched a number of electronic databases including CENTRAL, MEDLINE and Embase. Date last searched 2 September 2016. SELECTION CRITERIA We included randomised controlled trials (RCTs) in which adult participants had undergone surgery for age-related cataract. We included participants irrespective of their baseline risk of MO, in particular we included people with diabetes and uveitis. We included trials of preoperative and/or postoperative topical NSAIDs in conjunction with postoperative topical steroids. The comparator was postoperative topical steroids alone. A secondary comparison was preoperative and/or postoperative topical NSAIDs alone versus postoperative topical steroids alone. DATA COLLECTION AND ANALYSIS Two review authors independently selected studies for inclusion, assessed risk of bias and extracted data using standard methods expected by Cochrane. We pooled data using a random-effects model. We graded the certainty of the evidence using GRADE and considered the following: risk of bias of included studies, precision of the effect estimate, consistency of effects between studies, directness of the outcome measure and publication bias. MAIN RESULTS We identified 34 studies that were conducted in the Americas, Europe, the Eastern Mediterranean region and South-East Asia. Over 5000 people were randomised in these trials. The majority of studies enrolled one eye per participant; a small subset (4 trials) enrolled a proportion of people with bilateral surgery. Twenty-eight studies compared NSAIDs plus steroids with steroids alone. Six studies compared NSAIDs with steroids. A variety of NSAIDs were used, including ketorolac, diclofenac, nepafenac, indomethacin, bromfenac, flurbiprofen and pranopfen. Follow-up ranged from one to 12 months. In general, the studies were poorly reported. We did not judge any of the studies at low risk of bias in all domains. Six studies were funded by industry, seven studies were funded from non-industry sources, and the rest of the studies did not report the source of funding.There was low-certainty evidence that people receiving topical NSAIDs in combination with steroids may have a lower risk of poor vision due to MO at three months after cataract surgery compared with people receiving steroids alone (risk ratio (RR) 0.41, 95% confidence interval (CI) 0.23 to 0.76; eyes = 1360; studies = 5; I2 = 5%). We judged this to be low-certainty evidence because of risk of bias in the included studies and indirectness, as the extent of visual loss was not always clear. Only one study reported poor vision due to MO at 12 months and we judged this to be very low-certainty evidence as there were only two events. Quality of life was only reported in one of the 34 studies comparing NSAIDs plus steroids versus steroids alone, and it was not fully reported, other than to comment on lack of differences between groups. There was evidence of a reduced risk of MO with NSAIDs at three months after surgery, but we judged this to be low-certainty due to risk of bias and publication bias (RR 0.40, 95% CI 0.32 to 0.49; eyes = 3638; studies = 21). There was inconsistent evidence on central retinal thickness at three months (I2 = 87%). Results ranged from -30.9 µm in favour of NSAIDs plus steroids to 7.44 µm in favour of steroids alone. Similarly, data on best corrected visual acuity (BCVA) were inconsistent, but nine out of 10 trials reporting this outcome found between-group differences in visual acuity of less than 0.1 logMAR.None of the six studies comparing NSAIDs alone with steroids reported on poor vision due to MO at three or 12 months. There was low-certainty evidence that central retinal thickness was lower in the NSAIDs group at three months (mean difference (MD) -22.64 µm, 95% CI -38.86 to -6.43; eyes = 121; studies = 2). Five studies reported on MO and showed a reduced risk with NSAIDs, but we judged this evidence to be of low-certainty (RR 0.27, 95% CI 0.18 to 0.41; eyes = 520). Three studies reported BCVA at three months and the results of these trials were inconsistent, but all three studies found differences of less than 0.1 logMAR between groups.We did not note any major adverse events - the main consistent observation was burning or stinging sensation with the use of NSAIDs. AUTHORS' CONCLUSIONS Using topical NSAIDs may reduce the risk of developing macular oedema after cataract surgery, although it is possible that current estimates as to the size of this reduction are exaggerated. It is unclear the extent to which this reduction has an impact on the visual function and quality of life of patients. There is little evidence to suggest any important effect on vision after surgery. The value of adding topical NSAIDs to steroids, or using them as an alternative to topical steroids, with a view to reducing the risk of poor visual outcome after cataract surgery is therefore uncertain. Future trials should address the remaining clinical uncertainty of whether prophylactic topical NSAIDs are of benefit, particularly with respect to longer-term follow-up (at least to 12 months), and should be large enough to detect reduction in the risk of the outcome of most interest to patients, which is chronic macular oedema leading to visual loss.


title: [Clinical analysis of SARS: 27 cases report].

text: OBJECTIVE To realize the clinical characteristic of severe acute respiratory syndrome (SARS). METHODS Observe the clinical signs and symptoms of each patients; do laboratory examination periodically; and at last do Stat analysis of all the cases. RESULTS The first onset symptom of the 27 cases is high fever accompany with toxic symptom such as muscular soreness, headache, etc. The patient's condition may be most serious in the 2nd week. The amount of end-brush blood WBC decrease, especially the lymphocyte decrease more obviously. The amount of CD(4)(+) and CD(8)(+) decrease obviously is another important characteristic. The chest X-ray show pulmonary lesion in the 2nd day after onset of the illness, but 40.7% cases show positive change after 6 days onset. During the 7 - 12 days the pulmonary lesion become most serious. Seven cases are treated by steroid in the first 3 days when the chest X-ray is normal, but they become positive during 6 - 8 days of the course. Whether the state of the illness is serious or not seems no obvious relationship to when they are treated by steroid. And large dosage of steroid may inhibit the level of CD(4)(+) and CD8 (P < 0.05). Two severe cases present double infection after large dosage of steroid. CONCLUSION The course of a infectious atypical pneumonia is almost 3 weeks, the 1st week can be decided as early stage, the 2nd week is decided as fastigium, the 3rd week is decided as stage of recovery. During the fastigium, the disease transform very quickly, we need pay more attention to the patient. The CD(4)(+), CD(8)(+) and CD(3)(+) of SARS patient decrease obviously, it seems the immuno-function inhibited seriously. The serious case is easily accompany with severe secondary infection in the late stage, so the use of steroid must be carefully and the dosage need not to be much more.


Antiviral research at the Rega Institute (KU leuven), now 50 years old.


title: Outcome and characteristics of infants with acute viral bronchiolitis submitted to mechanical ventilation in a Brazilian pediatric intensive care.

text: OBJECTIVE To describe the characteristics and the outcome of infants with acute viral bronchiolitis submitted to mechanical ventilation. METHODS We performed a retrospective study enrolling all infants (less than 12 months old) admitted with the diagnosis of acute viral bronchiolitis and submitted to mechanical ventilation in an university affiliated Brazilian pediatric intensive care unit between March, 2004 and September, 2006 (3 consecutives winters). The mechanical ventilation parameters' employed on 1st, 2nd, 3rd, 7th day and before extubation were evaluated as well as the evolution (mortality rate, presence of acute respiratory distress syndrome and the prevalence of complications). The groups were compared using the Student t test, the Mann-Whitney U test and the Chi-square test. RESULTS Fifty-nine infants were included (3.8 ± 2.7 months old, 59% male), with 9.0 ± 9.4 days on mechanical ventilation. Prior mechanical ventilation, non invasive ventilation was instituted in 71% of children. Anemia was observed in 78% of the sample. In 51 infants (86.5%) the lower airway obstructive pattern was maintained up to tracheal extubation with a nil mortality and low prevalence of pneumothorax (7.8%). Acute respiratory distress syndrome occurred in 8 infants (13.5%), with higher mortality and a higher prevalence of pneumothorax (62.5%). CONCLUSIONS The declining mortality in acute viral bronchiolitis is observed even in non developed regions, involving children with high rates of anemia and premature labor. The low mortality is associated with the maintenance of the lower airway obstructive pattern during the period on mechanical ventilation. The development of acute respiratory distress syndrome is associated with increased mortality and higher prevalence of complications, representing the actual challenge in the management of children with severe acute viral bronchiolitis.


title: Mechanism of coronavirus transcription: duration of primary transcription initiation activity and effects of subgenomic RNA transcription on RNA replication.

text: Previously, we established a system whereby an intergenic region from mouse hepatitis virus (MHV) inserted into an MHV defective interfering (DI) RNA led to transcription of a subgenomic DI RNA in helper virus-infected cells. By using this system, the duration of a primary transcription initiation activity which transcribes subgenomic-size RNAs from the genomic-size RNA template in MHV-infected cells was examined. Efficient DI genomic and subgenomic RNA synthesis was observed when the DI RNA was transfected at 1, 3, 3.5, 5, and 6 h postinfection, indicating that all activities which are necessary for MHV RNA synthesis are present continuously during the first 6 h of infection. The effect of subgenomic DI RNA synthesis on DI genomic RNA replication was then examined. Replication efficiency of the DI genomic RNA which synthesized the subgenomic RNA was approximately 70% lower than that of DI genomic RNA which did not synthesize the subgenomic DI RNA in MHV-infected cells. Cotransfection of two different-size DI RNAs demonstrated that replication of the larger DI RNA was strongly inhibited by replication of the smaller genomic DI RNA. Cotransfection of two DI RNA species of the same length into MHV-infected cells demonstrated that reduced replication of the genomic DI RNA which synthesizes the subgenomic RNA did not affect the replication of cotransfected DI RNA, demonstrating that the reduction in DI genomic RNA replication works only in cis, not in trans. Therefore, the previously proposed hypothesis that coronavirus, subgenomic RNA synthesis may inhibit the replication of genomic RNA by competing for a limited amount of virus-derived factors seems unlikely. Possible mechanisms of coronavirus transcription are discussed.


title: Genes required for replication of the 15.5-kilobase RNA genome of a plant closterovirus.

text: A full-length cDNA clone of beet yellows closterovirus (BYV) was engineered and used to map functions involved in the replication of the viral RNA genome and subgenomic RNA formation. Among 10 open reading frames (ORFs) present in BYV, ORFs 1a and 1b suffice for RNA replication and transcription. The proteins encoded in these ORFs harbor putative methyltransferase, RNA helicase, and RNA polymerase domains common to Sindbis virus-like viruses and a large interdomain region that is unique to closteroviruses. The papain-like leader proteinase (L-Pro) encoded in the 5'-proximal region of ORF 1a was found to have a dual function in genome amplification. First, the autocatalytic cleavage between L-Pro and the remainder of the ORF 1a product was essential for replication of RNA. Second, an additional L-Pro function that was separable from proteolytic activity was required for efficient RNA accumulation. The deletion of a large, approximately 5.6-kb, 3'-terminal region coding for a 6-kDa hydrophobic protein, an HSP70 homolog, a 64-kDa protein, minor and major capsid proteins, a 20-kDa protein, and a 21-kDa protein (p21) resulted in replication-competent RNA. However, examination of mutants with replacements of start codons in each of these seven 3'-terminal ORFs revealed that p21 functions as an enhancer of genome amplification. The intriguing analogies between the genome organization and replicational requirements of plant closteroviruses and animal coronavirus-like viruses are discussed.


title: Sequence elements involved in the rescue of IBV defective RNA CD-91.

text: Deletion mutagenesis has been used to identify essential regions for rescue of coronavirus defective RNAs (D-RNAs). Using this technique on a cloned IBV D-RNA CD-91, we have identified a region potentially important in its rescue. Comparing the sequence of D-RNAs rescued with those not rescued we have deduced that a 72 base region corresponding to base number 13,824 to 13,896 in the viral genome is required for rescue. This may be an IBV D-RNA packaging signal or a cis-acting element involved in replication. Further experiments and modification of our techniques will be required to differentiate between the two processes.


title: Virological diagnosis in community-acquired pneumonia in immunocompromised patients.

text: Community-acquired pneumonia (CAP) is a serious lower respiratory tract infection associated with significant morbidity and mortality in immunocompromised patients. The present study evaluated the clinical spectrum of CAP in immunocompromised hosts and the role of respiratory viruses, as well as the yield of viral diagnostic methods. Conventional microbiological tests were routinely performed in immunocompromised patients with CAP. Nasopharyngeal swabs were processed for respiratory viruses by indirect immunofluorescence assay, cell culture and PCR. Four groups were defined according to aetiology of CAP, as follows: group 1 (nonviral), group 2 (mixed, nonviral and viral), group 3 (only viral) and group 4 (unknown aetiology). Over a 1-yr period, 92 patients were included. An aetiological diagnosis was achieved in 61 (66%) patients: 38 (41%), group 1; 12 (13%), group 2; and 11 (12%), group 3. The most frequent pathogen detected was Streptococcus pneumoniae (n = 29, 48%), followed by rhinovirus (n = 11, 18%). PCR identified 95% of respiratory viruses. Clinical characteristics could not reliably distinguish among the different aetiological groups. Respiratory viruses represent a substantial part of the aetiologies of community-acquired pneumonia in immunocompromised patients and its routine assessment through PCR in nasopharyngeal swabs should be considered in the clinical care of these patients.