title: Severe acute respiratory infection in children in a densely populated urban slum in Kenya, 2007–2011

text: BACKGROUND: Reducing acute respiratory infection burden in children in Africa remains a major priority and challenge. We analyzed data from population-based infectious disease surveillance for severe acute respiratory illness (SARI) among children <5 years of age in Kibera, a densely populated urban slum in Nairobi, Kenya. METHODS: Surveillance was conducted among a monthly mean of 5,874 (range = 5,778-6,411) children <5 years old in two contiguous villages in Kibera. Participants had free access to the study clinic and their health events and utilization were noted during biweekly home visits. Patients meeting criteria for SARI (WHO-defined severe or very severe pneumonia, or oxygen saturation <90%) from March 1, 2007-February 28, 2011 had blood cultures processed for bacteria, and naso- and oro- pharyngeal swabs collected for quantitative real-time reverse transcription polymerase chain reaction testing for influenza viruses, parainfluenza viruses (PIV), respiratory syncytial virus (RSV), adenovirus, and human metapneumovirus (hMPV). Swabs collected during January 1, 2009 – February 28, 2010 were also tested for rhinoviruses, enterovirus, parechovirus, Mycoplasma pneumoniae, and Legionella species. Swabs were collected for simultaneous testing from a selected group of control-children visiting the clinic without recent respiratory or diarrheal illnesses. RESULTS: SARI overall incidence was 12.4 cases/100 person-years of observation (PYO) and 30.4 cases/100 PYO in infants. When comparing detection frequency in swabs from 815 SARI cases and 115 healthy controls, only RSV and influenza A virus were significantly more frequently detected in cases, although similar trends neared statistical significance for PIV, adenovirus and hMPV. The incidence for RSV was 2.8 cases/100 PYO and for influenza A was 1.0 cases/100 PYO. When considering all PIV, the rate was 1.1 case/100 PYO and the rate per 100 PYO for SARI-associated disease was 1.5 for adenovirus and 0.9 for hMPV. RSV and influenza A and B viruses were estimated to account for 16.2% and 6.7% of SARI cases, respectively; when taken together, PIV, adenovirus, and hMPV may account for >20% additional cases. CONCLUSIONS: Influenza viruses and RSV (and possibly PIV, hMPV and adenoviruses) are important pathogens to consider when developing technologies and formulating strategies to treat and prevent SARI in children.


title: An Exit Strategy for Resuming Non-Emergency Neurosurgery after SARS-CoV-2: a UK Perspective.

text: Abstract Introduction Substantial healthcare resources have been diverted to deal with the effects of the Coronavirus SARS-CoV-2 pandemic, and non-emergency neurosurgery has been effectively shut down. As we start to emerge from the crisis, we need to deal with the backlog of non-emergency neurosurgical patients whose treatment has been delayed, whilst remaining responsive to further possible surges of infection. Aims & Method This article aims to identify core themes and challenges that limit resumption of a normal neurosurgical service after the SARS-CoV-2 pandemic, and to provide pragmatic advice and solutions that may be of utility to clinicians looking to resume non-emergency neurosurgical care. We reviewed of relevant international policies, a wide range of journalistic and media sources, and expert opinion to address the stated aims. Results We present and discuss a range of factors that may act as potential barriers to resuming full elective neurosurgical provision, along with important steps that must be overcome to achieve pre-COVID-19 surgical capacity. We also explore how we may overcome these challenges and outline key requirements for a successful neurosurgical exit strategy from the pandemic. Conclusion Recommencing non-emergency neurosurgery can start once minimum criteria have been fulfilled, these must include: 1. A structured prioritization of surgical cases 2. Incidence of virus infection should fall sufficiently to release previously diverted healthcare capacity 3. Adequate safety criteria met for patients and staff including sufficient personal protective equipment and robust testing 4. Maintenance of systems for rapid communication at organizational and individual levels


title: Preliminary studies on isolates of Clostridium difficile from dogs and exotic pets

text: BACKGROUND: Clostridium difficile infection (CDI) is recognised as an emerging disease in both humans and some animal species. During the past few years, insights into human CDI epidemiology changed and C. difficile is also considered as an emerging community-acquired pathogen. Certain ribotypes (RT) are possibly associated with zoonotic transmission. The objective of this study was to assess the presence of C. difficile in a population of pets and to characterise the isolates. RESULTS: Faecal samples from a total of 90 diarrhoeic dogs and 24 from exotic animal species (both diarrhoeic and non-diarrhoeic) were analysed. Clostridium difficile was isolated from 6 (6.7%) dogs and one reptile sample (4.2%). Four (66.7%) of the six dog strains were capable of producing toxins. Four known different RTs were detected in dogs (010, 014, 123 and 358) and a new one was found in a faecal sample of an exotic animal. This new RT isolate was negative for all toxin genes tested and belonged to sequence type 347 which has been proposed as a Clade-III member. Importantly, two dog strains showed a stable resistance to metronidazole (initial MIC values: 128 and 48 μg/ml). CONCLUSIONS: The results obtained in this study suggest the implementation of antimicrobial susceptibility surveillance programs to assess the prevalence of metronidazole resistance in dogs; molecular studies to elucidate C. difficile metronidazole resistance mechanisms are warranted. Based on the similarity between the ribotypes observed in dogs and those described in humans, the zoonotic transmission should be further explored. Furthermore, exotic animals have shown to harbor uncommon C. difficile strains which require further genomic studies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12917-018-1402-7) contains supplementary material, which is available to authorized users.


title: Anal swab findings in an infant with COVID‐19

text: INTRODUCTION: The transmission pathways of coronavirus disease 2019 (COVID‐19) remain not completely clear. In this case study the test for the novel severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) in pharyngeal swab and anal swab were compared. CASE PRESENTATION: A 3‐month‐old girl was admitted to our hospital with COVID‐19. Her parents had both been diagnosed with COVID‐19. The results of pharyngeal swab and anal swab of the little girl were recorded and compared during the course of the disease. The oropharyngeal specimen showed negative result for SARS‐CoV‐2 on the 14th day after onset of the illness. However, the anal swab was still positive for SARS‐CoV‐2 on the 28th day after the onset of the illness. CONCLUSION: The possibility of fecal‐oral transmission of COVID‐19 should be assessed. Personal hygiene during home quarantine merits considerable attention.


title: COVID-19 and Neurosurgery. Literature and Neurosurgical Societies Recommendations Update.

text: Abstract Background Since January 2020, 3 months ago, when the pathogen causing the Coronavirus-disease was identified in humans, the literature on COVID-19 has grown exponentially to over 4000 publications. There is the need to provide an update for each single medical discipline, including neurosurgery, to be used by single professionals or to be distributed through the neurosurgical community and to be used by governments in designing new scenario of care. Methods A review of the MEDLINE database was performed on April 13th, 2020. Search terms included “COVID-19”, “neurosurgery,” and “surgery”. A review of documents published on the web-page of the WFNS and of the 5 continental associations of neurosurgical societies AANS, AASNS, CAANS, EANS, and FLANC representing the 119 national Neurosurgical Societies around the world was performed. Results The literature search yielded 38 results that were manually reviewed. Fourteen manuscripts were considered eligible. They described suggestions and considerations to optimize care of neurosurgical patients, editorials on operational models, perspectives from neurosurgical departments, letters to the editor describing experiences on how to help medical staff to be prepared in advance for pandemic situations, description of regional or departmental models and/or organizational schemes. The webpages of the searched societies reported a total of 57 documents. Conclusions The neurosurgical scientific community has promptly reacted to the COVID-19 outbreak by producing a growing number of documents that could serve as guidance for neurosurgeons all over the world. Neurosurgical Societies will represent the key-institutions for guiding the neurosurgical community to overcome the COVID-19 crisis.


title: A virtual children's hospice in response to COVID-19: The Scottish experience

text: This case report describes a pediatric hospice provider in Scotland and their experience implementing a telehospice program in response to COVID-19. Children’s Hospices Across Scotland (CHAS) is the only provider of pediatric hospice care in the entire of Scotland, and we describe their experience offering pediatric telehospice. CHAS had strategically planned to implement telehospice, but COVID-19 accelerated the process. The organization evaluated its pediatric clinical and wrap-around hospice services and rapidly migrated them to a virtual environment. They creatively added new services to meet the unique needs of the entire family, who were caring for a child at end of life during COVID-19. CHAS’s experience highlights the planning and implementing processes of telehospice with key lessons learned, while acknowledging the challenges inherent in using technology to deliver hospice care.


title: Three Main Inducers of Alphacoronavirus Infection of Enterocytes: Sialic Acid, Proteases, and Low pH

text: Transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV) are similar coronaviruses, causing diseases characterized by vomiting, diarrhea, and death from severe dehydration in piglets. Thus, they have caused huge losses to the swine-breeding industry worldwide. Nowadays, they are easily transmitted among the continents via vehicles, equipment, and cargo. Both viruses establish an infection in porcine enterocytes in the small intestine, and their spike (S) proteins play a key role in the virus-cell binding process under unfavorable conditions when the intestine with a low pH is filled with a thick layer of mucus and proteases. Sialic acid, proteases, and low pH are three main inducers of coronavirus infection. However, the details of how sialic acid and low pH affect virus binding to the host cell are not determined, and the functions of the proteases are unknown. This review emphasizes the role of three factors in the invasion of TGEV and PEDV into porcine enterocytes and offers more insights into Alphacoronavirus infection in the intestinal environment.


title: Identification of putative polymerase gene product in cells infected with murine coronavirus A59

text: Abstract The virion RNA of mouse hepatitis virus, strain A59 (MHV-A59) is believed to be the mRNA for the viral RNA-dependent RNA polymerase. The cell-free translation of virion RNA results in the synthesis of two predominant products p220 and p28 (M. R. Denison and S. Perlman, 1986, J. Virol. 60, 12–18). p28 is a basic protein and is readily detected by two-dimensional gel electrophoresis. When infected cells and isolated virions were assayed for this protein by two-dimensional gel electrophoresis, p28 could be detected in infected cells labeled at late times after infection, but not at early times or in purified virions. p28 represents the first protein product of the putative coronavirus polymerase gene to be identified in infected cells.


title: Effect of cinnamon water extract on monocyte-to-macrophage differentiation and scavenger receptor activity

text: BACKGROUND: Water soluble cinnamon extract has been shown to increase insulin sensitivity and modulate macrophage activation, a desirable trait for the management of obesity or atherosclerosis. Our present study investigated whether cinnamon water extract (CWE) may influence the differentiation of monocytes into macrophages and the activity of macrophage scavenger receptors, commonly observed in atherosclerotic lesions. METHODS: We investigated the effect of CWE on the expression of various surface markers and the uptake of acetylated low density lipoprotein (LDL) in phorbol-12-myristate-13-acetate (PMA)-stimulated THP-1 cells. The protein levels of PMA or macrophage-colony stimulating factor (M-CSF)-stimulated type 1 macrophage scavenger receptor (SRA) were analyzed. Finally, the role of extracellar signal-related kinase (ERK) 1/2 in SRA synthesis and the effect of CWE on PMA-stimulated ERK1/2 were determined. RESULTS: CWE inhibited the differentiation of monocyte by decreasing the expression of CD11b, CD36 and SRA and the uptake of acetyl LDL. CWE suppressed the upregulation of SRA by M-CSF and modulated ERK1/2 activity, which was required for PMA-induced SRA synthesis. CONCLUSIONS: Our results demonstrate that CWE was able to interfere with monocyte differentiation and macrophage scavenger activity, indicating its potential in preventing the development of atherosclerotic lesions.


title: Molecular mechanism of evolution and human infection with the novel coronavirus (2019-nCoV)

text: Since December, 2019, an outbreak of pneumonia caused by the new coronavirus (2019-nCoV) has hit the city of Wuhan in the Hubei Province. With the continuous development of the epidemic, it has become a national public health crisis and calls for urgent antiviral treatments or vaccines. The spike protein on the coronavirus envelope is critical for host cell infection and virus vitality. Previous studies showed that 2019-nCoV is highly homologous to human SARS-CoV and attaches host cells though the binding of the spike receptor binding domain (RBD) domain to the angiotensin-converting enzyme II (ACE2). However, the molecular mechanisms of 2019-nCoV binding to human ACE2 and evolution of 2019-nCoV remain unclear. In this study, we have extensively studied the RBD-ACE2 complex, spike protein, and free RBD systems of 2019-nCoV and SARS-CoV using protein-protein docking and molecular dynamics (MD) simulations. It was shown that the RBD-ACE2 binding free energy for 2019-nCoV is significantly lower than that for SARS-CoV, which is consistent with the fact that 2019-nCoV is much more infectious than SARS-CoV. In addition, the spike protein of 2019-nCoV shows a significantly lower free energy than that of SARS-CoV, suggesting that 2019-nCoV is more stable and able to survive a higher temperature than SARS-CoV. This may also provide insights into the evolution of 2019-nCoV because SARS-like coronaviruses are thought to have originated in bats that are known to have a higher body-temperature than humans. It was also revealed that the RBD of 2019-nCoV is much more flexible especially near the binding site and thus will have a higher entropy penalty upon binding ACE2, compared to the RBD of SARS-CoV. That means that 2019-nCoV will be much more temperature-sensitive in terms of human infection than SARS-CoV. With the rising temperature, 2019-nCoV is expected to decrease its infection ability much faster than SARS-CoV, and get controlled more easily. The present findings are expected to be helpful for the disease prevention and control as well as drug and vaccine development of 2019-nCoV.


title: Efficient assembly and release of SARS coronavirus-like particles by a heterologous expression system

text: Virus-like particles (VLPs) produced by recombinant expression of the major viral structural proteins could be an attractive method for severe acute respiratory syndrome (SARS) control. In this study, using the baculovirus system, we generated recombinant viruses that expressed S, E, M and N structural proteins of SARS-CoV either individually or simultaneously. The expression level, size and authenticity of each recombinant SARS-CoV protein were determined. In addition, immunofluorescence and FACS analysis confirmed the cell surface expression of the S protein. Co-infections of insect cells with two recombinant viruses demonstrated that M and E could assemble readily to form smooth surfaced VLPs. On the other hand, simultaneous high level expression of S, E and M by a single recombinant virus allowed the very efficient assembly and release of VLPs. These data demonstrate that the VLPs are morphological mimics of virion particles. The high level expression of VLPs with correct S protein conformation by a single recombinant baculovirus offers a potential candidate vaccine for SARS.


title: Reprogramming the unfolded protein response for replication by porcine reproductive and respiratory syndrome virus

text: The unfolded protein response (UPR) in the endoplasmic reticulum (ER) constitutes a critical component of host innate immunity against microbial infections. In this report, we show that porcine reproductive and respiratory syndrome virus (PRRSV) utilizes the UPR machinery for its own benefit. We provide evidence that the virus targets the UPR central regulator GRP78 for proteasomal degradation via a mechanism that requires viral glycoprotein GP2a, while both IRE1-XBP1s and PERK-eIF2α-ATF4 signaling branches of the UPR are turned on at early stage of infection. The activated effector XBP1s was found to enter the nucleus, but ATF4 was unexpectedly diverted to cytoplasmic viral replication complexes by means of nonstructural proteins nsp2/3 to promote viral RNA synthesis. RNAi knockdown of either ATF4 or XBP1s dramatically attenuated virus titers, while overexpression caused increases. These observations reveal attractive host targets (e.g., ATF4 and XBP1s) for antiviral drugs and have implications in vaccine development.


Comment on “Beware of Too Aggressive Approach in Children With Acute Abdomen During COVID-19 Outbreak!”


title: Characterization of the immune responses elicited by baculovirus-based vector vaccines against influenza virus hemagglutinin

text: AIM: To compare the specific immune responses elicited by different baculovirus vectors in immunized mice. METHODS: We constructed and characterized two recombinant baculoviruses carrying the expression cassette for the H5N1 influenza virus hemagglutinin (HA) gene driven by either an insect cell promoter (vAc-HA) or a dual-promoter active both in insect and mammalian cells (vAc-HA-DUAL). Virus without the HA gene (vAc-EGFP) was used as a control. These viruses were used to immunize mice subcutaneously and intraperitoneally. The production of total and specific antibodies was determined by ELISA and competitive ELISA. Cytokine production by the spleen cells of immunized mice was studied using the ELISPOT assay. RESULTS: Both the vAc-HA and vAc-HA-DUAL vectors expressed HA proteins in insect Sf9 cells, and HA antigen was present in progeny virions. The vAc-HA-DUAL vector also mediated HA expression in virus-transduced mammalian cell lines (BHK and A547). Both vAc-HA and vAc-HA-DUAL exhibited higher transduction efficiencies than vAc-EGFP in mammalian cells, as shown by the expression of the reporter gene egfp. Additionally, both vAc-HA and vAc-HA-DUAL induced high levels of HA-specific antibody production in immunized mice; vAc-HA-DUAL was more efficient in inducing IFN-γ and IL-2 upon stimulation with specific antigen, whereas vAc-HA was more efficient in inducing IL-4 and IL-6. CONCLUSION: Baculovirus vectors elicited efficient, specific immune responses in immunized mice. The vector displaying the HA antigen on the virion surface (vAc-HA) elicited a Th2-biased immune response, whereas the vector displaying HA and mediating HA expression in the cell (vAc-HA-DUAL) elicited a Th1-biased immune response.


title: Sustainable practice of ophthalmology during COVID-19: challenges and solutions

text: PURPOSE: The Coronavirus (COVID-19) outbreak is rapidly emerging as a global health threat. With no proven vaccination or treatment, infection control measures are paramount. In this article, we aim to describe the impact of COVID-19 on our practice and share our strategies and guidelines to maintain a sustainable ophthalmology practice. METHODS: Tan Tock Seng Hospital (TTSH) Eye Centre is the only ophthalmology department supporting the National Centre for Infectious Diseases (NCID), which is the national screening center and the main center for management of COVID-19 patients in Singapore. Our guidelines during this outbreak are discussed. RESULTS: Challenges in different care settings in our ophthalmology practice have been identified and analyzed with practical solutions and guidelines implemented in anticipation of these challenges. First, to minimize cross-infection of COVID-19, stringent infection control measures were set up. These include personal protective equipment (PPE) for healthcare workers and routine cleaning of “high-touch” surfaces. Second, for outpatient care, a stringent dual screening and triaging process were carried out to identify high-risk patients, with proper isolation for such patients. Administrative measures to lower patient attendance and reschedule appointments were carried out. Third, inpatient and outpatient care were separated to minimize interactions. Last but not least, logistics and manpower plans were drawn up in anticipation of resource demands and measures to improve the mental well-being of staff were implemented. CONCLUSION: We hope our measures during this COVID-19 pandemic can help ophthalmologists globally and serve to guide and maintain safe access in ophthalmology clinics when faced with similar disease outbreaks.


title: Examining professional emergency managers in Korea

text: Abstract Although the number of emergency managers has risen in South Korea (hereafter referred to as Korea) over the years, their role is not yet as defined and noteworthy compared to other professions because of its unidisciplinary approach. This article investigates how Korea has to improve emergency managers' disciplinary approach to ultimately contribute to the goal of effective transnational disaster management. This study uses qualitative content analysis of government policies, college curricula, nongovernmental organizations' (NGOs') emergency-manager certification, and mass media coverage to compare emergency managers' unidisciplinary and multidisciplinary approaches. The key tenet is that Korea must change its emergency managers' unidisciplinary approach into a multidisciplinary approach because the former is less effective when dealing with complicated disaster management systems. To achieve this change, the stakeholders must carry out their assigned responsibilities under risk-oriented management. As for the study's international implications, developing nations may consider the enhancement of related educational curricula, collaborative learning, continuous evaluation, disaster awareness, and disaster prevention for the emergency managers' multidisciplinary approach.


title: Pandemic stricken cities on lockdown. Where are our planning and design professionals [now, then and into the future]?

text: Chinese cities have been placed upon lockdown in early 2020 in an attempt to contain the novel coronavirus (COVID-19), as increasingly huge demands are being placed upon Chinese and international health professionals to address this pandemic. Surprisingly, planning and design professionals are absent in the discourses about existing and post−COVID-19 strategies and actions even though previous pandemics historically revealed major impacts on the urban fabric from social and economic perspectives. This paper is a call for action for international architectural and urban organisations to include pandemics and similar in their disaster management strategies. This need is very evident in their need to better design creative and relevant protocols in partnership with health discipine organisations, and so that their applied deployment in pandemic stricken cities can be effected integrated seamlessly within normal city environment planning activities and also in incident situations like containing the current COVID-19 pandemic.


title: C-ME: A 3D Community-Based, Real-Time Collaboration Tool for Scientific Research and Training

text: The need for effective collaboration tools is growing as multidisciplinary proteome-wide projects and distributed research teams become more common. The resulting data is often quite disparate, stored in separate locations, and not contextually related. Collaborative Molecular Modeling Environment (C-ME) is an interactive community-based collaboration system that allows researchers to organize information, visualize data on a two-dimensional (2-D) or three-dimensional (3-D) basis, and share and manage that information with collaborators in real time. C-ME stores the information in industry-standard databases that are immediately accessible by appropriate permission within the computer network directory service or anonymously across the internet through the C-ME application or through a web browser. The system addresses two important aspects of collaboration: context and information management. C-ME allows a researcher to use a 3-D atomic structure model or a 2-D image as a contextual basis on which to attach and share annotations to specific atoms or molecules or to specific regions of a 2-D image. These annotations provide additional information about the atomic structure or image data that can then be evaluated, amended or added to by other project members.


title: Innovation in Response to the COVID-19 Pandemic Crisis

text: The COVID-19 pandemic has disrupted all aspects of academic medical center missions. The number and rapidity of innovative responses to the crisis are extraordinary. When the pandemic has subsided, the world of academic medicine will have changed. The author of this Invited Commentary anticipates that at least some of these innovations will become part of academic medicine’s everyday clinical and educational operations. Here, he considers the implications of exemplary innovations—virtual care, hospital at home, advances in diagnosis and therapy, virtual learning, and virtual clinical learning—for regulators, academic medical centers, faculty, and students.


The Dermatologist’s Perspective: Why is COVID-19 mortality lower in females than males?


title: Initial psychological impact of COVID-19 and its correlates in Indian Community: An online (FEEL-COVID) survey

text: BACKGROUND: The pandemic of Corona Virus (COVID-19) hit India recently; and the associated uncertainty is increasingly testing psychological resilience of the masses. When the global focus has mostly been on testing, finding a cure and preventing transmission; people are going through a myriad of psychological problems in adjusting to the current lifestyles and fear of the disease. Since there is a severe dearth of researches on this issue, we decided to conduct an online survey to evaluate its psychological impact. METHODS: From 26(th) to 29(th) March an online survey (FEEL-COVID) was conducted using principles of snowballing, and by invitation through text messages to participate. The survey collected data on socio-demographic and clinical variables related to COVID-19 (based on the current knowledge); along with measuring psychological impact with the help of Impact of Event–revised (IES-R) scale. RESULTS: There were a total of 1106 responses from around 64 cities in the country. Out of these 453 responses had at least one item missing; and were excluded from the analysis. The mean age of the respondents was around 41 years with a male female ratio of 3:1 and around 22% respondents were health care professionals. Overall approximately one third of respondents had significant psychological impact (IES-R score > 24). Higher psychological impact was predicted with younger age, female gender and comorbid physical illness. Presence of physical symptoms and contact history predicted higher psychological impact, but did not reach statistical significance. CONCLUSION: During the initial stages of COVID-19 in India, almost one-third respondents had a significant psychological impact. This indicates a need for more systematic and longitudinal assessment of psychological needs of the population, which can help the government in formulating holistic interventions for affected individuals.


title: De l’intime au collectif, de multi-traumatismes sont en cours: Osons prévenir plutôt que guérir, osons une politique éthique

text: Résumé L’épidémie de Covid-19, sa mortalité, les incertitudes qui persistent sur son origine, son évolution, la période de confinement et ses suites entraînent des situations d’inquiétude voire d’angoisse jusqu'au traumatisme pour certains. Si celles-ci sont naturellement légitimes, cela ne signifie pas qu’elles ne doivent pas être prises au sérieux, et bénéficier d’une prise en charge appropriée dès lors qu’elles impactent significativement la vie des individus, sachant que la prévention reste le meilleur des soins. Abstract The Covid-19 epidemic, its mortality, the uncertainties that persist on its origin, its evolution, the period of confinement and its consequences lead to situations of anxiety and even anguish to the point of trauma for some people. While these are naturally legitimate, this does not mean that they should not be taken seriously, and benefit from appropriate care as soon as they have a significant impact on the lives of individuals, knowing that prevention remains the best form of care.


title: Management of heart transplant candidates in the time of COVID‐19 pandemic: Looking for answers

text: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-associated coronavirus disease 2019 (COVID-19) pandemic is dramatically challenging the healthcare system. In the absence of optimal testing and effective treatments, physicians are forced to make decisions relying on scarce or even controversial evidence. Setting of solid organ transplant (SOT) candidates and recipients offers clinical and organizational issues.


title: Arterivirus nsp12 versus the coronavirus nsp16 2′-O-methyltransferase: comparison of the C-terminal cleavage products of two nidovirus pp1ab polyproteins

text: The 3′-terminal domain of the most conserved ORF1b in three of the four families of the order Nidovirales (except for the family Arteriviridae) encodes a (putative) 2′-O-methyltransferase (2′-O-MTase), known as non structural protein (nsp) 16 in the family Coronaviridae and implicated in methylation of the 5′ cap structure of nidoviral mRNAs. As with coronavirus transcripts, arterivirus mRNAs are assumed to possess a 5′ cap although no candidate MTases have been identified thus far. To address this knowledge gap, we analysed the uncharacterized nsp12 of arteriviruses, which occupies the ORF1b position equivalent to that of the nidovirus 2′-O-MTase (coronavirus nsp16). In our in-depth bioinformatics analysis of nsp12, the protein was confirmed to be family specific whilst having diverged much further than other nidovirus ORF1b-encoded proteins, including those of the family Coronaviridae. Only one invariant and several partially conserved, predominantly aromatic residues were identified in nsp12, which may adopt a structure with alternating α-helices and β-strands, an organization also found in known MTases. However, no statistically significant similarity was found between nsp12 and the twofold larger coronavirus nsp16, nor could we detect MTase activity in biochemical assays using recombinant equine arteritis virus (EAV) nsp12. Our further analysis established that this subunit is essential for replication of this prototypic arterivirus. Using reverse genetics, we assessed the impact of 25 substitutions at 14 positions, yielding virus phenotypes ranging from WT-like to non-viable. Notably, replacement of the invariant phenylalanine 109 with tyrosine was lethal. We concluded that nsp12 plays an essential role during EAV replication, possibly by acting as a co-factor for another enzyme.


title: Characterizations of Coronaviruscis-Acting RNA Elements and the Transcription Step Affecting Its Transcription Efficiency

text: Abstract Seven to eight species of viral subgenomic mRNAs are produced in coronavirus-infected cells. These mRNAs are produced in different quantities, and their molar ratios remain constant during viral replication. We studied RNA elements that affect coronavirus transcription efficiency by characterizing a series of cloned coronavirus mouse hepatitis virus (MHV) defective interfering (DI) RNAs containing an inserted intergenic sequence, from which subgenomic DI RNA is transcribed in MHV-infected cells. Certain combinations of upstream and downstream flanking sequences of the intergenic sequence suppressed subgenomic DI RNA transcription, yet changing one of the flanking sequences to a different sequence eliminated transcription suppression. The suppressive effect of certain combinations of flanking sequences, but not all combinations, could be counteracted by altering the intergenic sequence. Thus, the combination of intergenic sequence and flanking sequence affected transcription efficiency. We also characterized another set of DI RNAs designed to clarify which transcription step determines the relative molar ratios of coronavirus mRNAs. Our study indicated that if subgenomic mRNAs were exclusively synthesized from negative-strand genomic RNA, then the relative molar ratios of coronavirus mRNAs were most likely determined after synthesis of the genomic-sized template RNA. If negative-strand subgenomic RNAs were templates for subgenomic mRNAs, then the relative molar ratios of coronavirus mRNAs probably were determined after synthesis of the genomic-sized template RNA used for subgenomic-sized RNA transcription but prior to the completion of the synthesis of subgenomic-sized RNAs containing the leader sequence. The relative molar ratios of coronavirus mRNAs, therefore, seem to have been established prior to a putative replicon-type amplification of subgenomic mRNAs.


SARS transmission on an aircraft


title: The permeabilized SecY protein-translocation channel can serve as a nonspecific sugar transporter

text: As the initial step in carbohydrate catabolism in cells, the substrate-specific transporters via active transport and facilitated diffusion play a decisive role in passage of sugars through the plasma membrane into the cytoplasm. The SecY complex (SecYEG) in bacteria forms a membrane channel responsible for protein translocation. This work demonstrates that weakening the sealability of the SecY channel allowed free diffusion of sugars, including glucose, fructose, mannose, xylose, arabinose, and lactose, into the engineered cells, facilitating its rapid growth on a wide spectrum of monosaccharides and bypassing/reducing stereospecificity, transport saturation, competitive inhibition, and carbon catabolite repression (CCR), which are usually encountered with the specific sugar transporters. The SecY channel is structurally conserved in prokaryotes, thus it may be engineered to serve as a unique and universal transporter for bacteria to passage sugars as demonstrated in Escherichia coli and Clostridium acetobutylicum.


title: Genetic diversity of a canine coronavirus detected in pups with diarrhoea in Italy

text: Abstract The sequence of the S gene of a field canine coronavirus (CCoV), strain Elmo/02, revealed low nucleotide (61%) and amino acid (54%) identity to reference CCoV strains. The highest correlation (77% nt and 81.7% aa) was found with feline coronavirus type I. A PCR assay for the S gene of strain Elmo/02 detected analogous CCoVs of different geographic origin, all which exhibited at least 92–96% nucleotide identity to each other and to strain Elmo/02. The evident genetic divergence between the reference CCoV strains and the newly identified Elmo/02-like CCoVs strongly suggests that a novel genotype of CCoV is widespread in the dog population.


title: IFN‐λ4 inhibits HIV infection of macrophages through signalling of IFN‐λR1/IL‐10R2 receptor complex

text: The recently discovered IFN‐λ4 has been found to have antiviral activity against several viruses. However, it's unknown whether IFN‐λ4 can inhibit HIV infection. Here, we show that IFN‐λ4 could suppress HIV infection of macrophages. This IFN‐λ4‐mediated HIV inhibition was compromised by the antibodies against IFN‐λ receptor complex, IFN‐λR1/IL‐10R2. IFN‐λ4 enhanced the phosphorylation of STAT1, and induced antiviral interferon‐stimulated genes. These findings indicated that IFN‐λ4 can inhibit HIV via JAK/STAT signalling pathway.


title: Molecular dynamics simulations of viral RNA polymerases link conserved and correlated motions of functional elements to fidelity

text: The viral RNA-dependent RNA polymerase (RdRp) is essential for multiplication of all RNA viruses. The sequence diversity of an RNA virus population contributes to its ability to infect the host. This diversity emanates from errors made by the RdRp during RNA synthesis. The physical basis for RdRp fidelity is unclear but is linked to conformational changes occurring during the nucleotide-addition cycle. To understand RdRp dynamics that might influence RdRp function, we have analyzed all-atom molecular dynamics (MD) simulations on the nanosecond timescale of four RdRps from the picornavirus family that exhibit 30–74% sequence identity. Principal component analysis showed that the major motions observed during the simulations derived from conserved structural motifs and regions of known function. Dynamics of residues participating in the same biochemical property, for example RNA binding, nucleotide binding or catalysis, were correlated even when spatially distant on the RdRp structure. The conserved and correlated dynamics of functional, structural elements suggest co-evolution of dynamics with structure and function of the RdRp. Crystal structures of all picornavirus RdRps exhibit a template-nascent RNA duplex channel too small to fully accommodate duplex RNA. Simulations revealed opening and closing motions of the RNA and NTP channels, which might be relevant to NTP entry, PP(i) exit and translocation. A role for nanosecond timescale dynamics in RdRp fidelity is supported by altered dynamics of the high-fidelity G64S derivative of PV RdRp relative to wild-type enzyme.


title: How did we get here? A short history of COVID‐19 and other coronavirus‐related epidemics

text: The COVID‐19 epidemic was not the first coronavirus epidemic of this century and represents one of the increasing number of zoonoses from wildlife to impact global health. SARS CoV‐2, the virus causing the COVID‐19 epidemic is distinct from, but closely resembles SARS CoV‐1, which was responsible for the severe acute respiratory syndrome (SARS) outbreak in 2002. SARS CoV‐1 and 2 share almost 80% of genetic sequences and use the same host cell receptor to initiate viral infection. However, SARS predominantly affected individuals in close contact with infected animals and health care workers. In contrast, CoV‐2 exhibits robust person to person spread, most likely by means of asymptomatic carriers, which has resulted in greater spread of disease, overall morbidity and mortality, despite its lesser virulence. We review recent coronavirus‐related epidemics and distinguish clinical and molecular features of CoV‐2, the causative agent for COVID‐19, and review the current status of vaccine trials.


title: Chapter Twelve Application of MALDI-TOF Mass Spectrometry in Clinical Virology

text: Abstract Matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF MS) is a diagnostic tool of microbial identification and characterization based on the detection of the mass of molecules. In the majority of clinical laboratories, this technology is currently being used mainly for bacterial diagnosis, but several approaches in the field of virology have been investigated. The introduction of this technology in clinical virology will improve the diagnosis of infections produced by viruses but also the discovery of mutations and variants of these microorganisms as well as the detection of antiviral resistance. This chapter is focused on the main current applications of MALDI-TOF MS techniques in clinical virology showing the state of the art with respect to this exciting new technology.


title: COVID-19 Pandemic: A Time for Collaboration and A Unified Global Health Front

text: Coronavirus disease 2019 (COVID-19) has, in the span of weeks, immobilized entire countries and mobilized leading institutions worldwide in a race towards treatments and preventions. While several solutions such as telemedicine and online education platforms have been implemented to reduce human contact and further transmission, countries need to favour collectivism both within and beyond their borders. Inspired by experiences of previous outbreaks such as SARS in 2003 and Ebola in 2014–2015, global solidarity is a must in order to prevent further morbidity and mortality. Examples in leadership and collaborations ranging from research funds from the Bill and Melinda Gates Foundation to mask donations by the Jack Ma Foundation should be celebrated as examples to follow. Open communication and transparency will be crucial in monitoring the evolution of the disease in the global effort of flattening the curve. This crisis will challenge the integrity and fuel innovation of health systems worldwide, whilst posing a new quality chasm that warrants increased recognition.


COVID-19 crisis: an extraordinary time for collaboration and science—a call for leadership, now and beyond


title: Recessions and health: The long‐term health consequences of responses to the coronavirus

text: The lockdown measures that were implemented in the spring of 2020 to stop the spread of Covid‐19 are having a huge impact on economies in the UK and around the world. In addition to the direct impact of Covid‐19 on health, the following recession will have an impact on people's health outcomes. This paper reviews economic literature on the longer run health impacts of business cycle fluctuations and recessions. Previous studies show that an economic downturn, which affects people through increased unemployment, lower incomes and increased uncertainty, will have significant consequences on people's health outcomes both in the short and longer term. The health effects caused by these adverse macroeconomic conditions will be complex, and will differ across generations, regions and socioeconomic groups. Groups that are vulnerable to poor health are likely to be hit hardest even if the crisis hit all individuals equally, and we already see that some groups such as young workers and women are worse hit by the recession than others. Government policies during and after the pandemic will play an important role in determining the eventual health consequences. This article is protected by copyright. All rights reserved


title: Audience readings of media messages about MRSA

text: Summary This paper explores whether, and to what extent, national newspaper messages tally with public perceptions about meticillin-resistant Staphylococcus aureus (MRSA). It compares research on media messages about MRSA with interview data gathered from a demographically diverse sample of 60 people interviewed from the Greater London area. Across the interview sample there was a shared consensus that most people associated MRSA not with the history of antibiotic use, but with dirty and poorly managed hospitals. Some media messages, such as blaming MRSA on the alleged ‘management culture’ of the NHS, seemed to capture the Zeitgeist, whereas others, in particular the ‘celebrity victims’ of MRSA, did not seem to resonate with the audience. This study also found that ideas based on scientific understandings about germ theory and the immune system were held alongside folklore such as miasmic theory. The comparison of media and mind thus points to the existence of pre-scientific understandings of germs, contagion and blame in parallel with the biomedical story in the minds of the public. The findings contribute to our understanding of the public and patients' views of this infection.


title: Prevalence of Cryptosporidium infection in calves in France

text: Abstract Two multicentre surveys were conducted in France to estimate the prevalence of Cryptosporidium infection in calves using qualitative ELISA for detection of Cryptosporidium coproantigens and oocysts. The first survey involved 4–12-day-old calves in six dairy-calf distribution centres, collecting calves from seven Administrative Regions (Aquitaine, Bretagne, Franche-Comté, Lorraine, Normandie, Nord, Pays de Loire). For each region, 20 calves were selected every month for 12 consecutive months (October 1995–September 1996). Prevalence of Cryptosporidium infection was 17.9% (Confidence Intervals (C.I.) 95%=[16.1%; 19.8%]) among the 1628 selected calves, of which only 5.3% had diarrhoea. The second survey conducted between November 1995 and May 1996 involved 4–21-day-old calves examined by veterinary practitioners who selected 189 livestock farms of dairy- or suckler-type in ten Administrative Departments (Allier, Cantal, Creuse, Doubs, Ille-et-Vilaine, Maine-et-Loire, Manche, Pas-de-Calais, Saône-et-Loire, Vendée). Cryptosporidia were detected in 105 (55.6%) of the farms. Among the 440 calves examined, of which 398 (90.5%) presented diarrhoea, cryptosporidia were found in 191 animals, i.e. a prevalence of 43.4% (C.I. 95%=[38.8%; 48.0%]). Breed of calves and type of housing had very little impact on prevalence in this survey. Some regional variations could be noticed, even if cryptosporidia infection is widespread. Monthly variations could be related to seasonal peaks in calving with a lower infection rate during summer.


title: An analysis of imported infections over a 5-year period at a teaching hospital in the United Kingdom

text: Abstract Background. Imported infections are an important cause of morbidity and mortality in the United Kingdom. Methods. A 5-year analysis of cases seen in a large teaching and district general hospital in the Eastern Region of the UK was performed using ward records correlated with Hospital coding data and Hospital Episode Statistics from the Department of Health. Results. A surprising number (301) and diversity of imported infections was diagnosed. Prophylactic measures were, where assessable, generally inadequate. Conclusions. These data warrant renewed efforts to educate travellers of the risks of infection acquired abroad. The continued rise in global travel along with emergence of new infectious diseases emphasises further the need for expanded infectious diseases services incorporating accessible travel advice services in the UK which are currently underprovided.


title: Mesenchymal Stem Cells for Coronavirus (COVID-19)-Induced Pneumonia: Revisiting the Paracrine Hypothesis with New Hopes?

text: Mesenchymal stem cells (MSCs) bear a promising potential for regenerative medicine therapies and they repair damaged tissue through secretion of immune modulatory and anti-inflammatory molecules acting in a paracrine fashion. Coronavirus disease 2019 (COVID-19) has spread all over the world with high morbidity and mortality rates and there is no specific treatment for this infection. A recent study published in the journal reports that MSC infusion is safe and effective in patients suffering from COVID-19 induced pneumonia. In the light of this study and previous reports, we make additional comments about possible therapeutic effects of MSCs in COVID-19 infection.


title: Safe tracheostomy for patients with severe acute respiratory syndrome

text: Objectives/Hypothesis: Severe acute respiratory syndrome (SARS) caused by coronavirus has become an epidemic affecting many regions worldwide. Fourteen percent to 20% of patients require endotracheal intubation and ventilator support. Some of these patients may require tracheostomy subsequently. This procedure, when performed without protection, may lead to infection of the medical and nursing staff taking care of the patient. Study Design: Based on clinical information of three patients. Methods: The authors carried out an emergency tracheostomy and changed the tracheostomy tube for one patient and performed elective tracheostomy in another two patients. Results: No medical or nursing staff member was infected after carrying out the procedure while taking all the precautions and wearing the appropriate protective apparel. Conclusion: The authors have prepared guidelines for performing a safe tracheostomy under both elective and emergency conditions. Surgeons who might be involved in performing the tracheostomy should become familiar with these guidelines and the appropriate protective apparel.


Intensive Care Unit Admissions for Pregnant and Non-Pregnant Women with COVID-19


title: COVID-19: Challenges to GIS with Big Data

text: Abstract The outbreak of the 2019 novel coronavirus disease (COVID-19) has caused more than 100,000 people infected and thousands of deaths. Currently, the number of infections and deaths is still increasing rapidly. COVID-19 seriously threatens human health, production, life, social functioning and international relations. In the fight against COVID-19, Geographic Information Systems (GIS) and big data technologies have played an important role in many aspects, including the rapid aggregation of multi-source big data, rapid visualization of epidemic information, spatial tracking of confirmed cases, prediction of regional transmission, spatial segmentation of the epidemic risk and prevention level, balancing and management of the supply and demand of material resources, and social-emotional guidance and panic elimination, which provided solid spatial information support for decision-making, measures formulation, and effectiveness assessment of COVID-19 prevention and control. GIS has developed and matured relatively quickly and has a complete technological route for data preparation, platform construction, model construction, and map production. However, for the struggle against the widespread epidemic, the main challenge is finding strategies to adjust traditional technical methods and improve speed and accuracy of information provision for social management. At the data level, in the era of big data, data no longer come mainly from the government but are gathered from more diverse enterprises. As a result, the use of GIS faces difficulties in data acquisition and the integration of heterogeneous data, which requires governments, businesses, and academic institutions to jointly promote the formulation of relevant policies. At the technical level, spatial analysis methods for big data are in the ascendancy. Currently and for a long time in the future, the development of GIS should be strengthened to form a data-driven system for rapid knowledge acquisition, which signifies that GIS should be used to reinforce the social operation parameterization of models and methods, especially when providing support for social management.


title: Activation of Notch signaling during ex vivo expansion maintains donor muscle cell engraftment

text: Transplantation of myogenic stem cells possesses great potential for long-term repair of dystrophic muscle. However, a single donor muscle biopsy is unlikely to provide enough cells to effectively transplant the muscle mass of a patient affected by muscular dystrophy. Expansion of cells ex vivo using traditional culture techniques significantly reduces engraftment potential. We hypothesized that activation of Notch signaling during ex vivo expansion would maintain donor cell engraftment potential. In this study, we expanded freshly isolated canine muscle-derived cells on tissue culture plates coated with Delta-1(ext)-IgG to activate Notch signaling or with human IgG as a control. A model of canine-to-murine xenotransplantation was used to quantitatively compare canine muscle cell engraftment, and determine if engrafted donor cells could function as satellite cells in vivo. We show that Delta-1(ext)-IgG inhibited differentiation of canine muscle-derived cells, and increased the level of genes normally expressed in myogenic precursors. Moreover, cells expanded on Delta-1(ext)-IgG resulted in a significant increase in the number of donor-derived fibers, as compared to cells expanded on human IgG, reaching engraftment levels similar to freshly isolated cells. Importantly, cells expanded on Delta-1(ext)-IgG engrafted to the recipient satellite cell niche, and contributed to further regeneration. A similar strategy of expanding human muscle-derived cells on Notch ligand might facilitate engraftment and muscle regeneration for patients affected with muscular dystrophy.


title: An overview on COVID-19: reality and expectation

text: Recently, severe acute respiratory syndrome coronavirus 2 (SARS-COV-2), commonly known as coronavirus disease-2019 (COVID-19) has rapidly spread across China and around the world. By the declaration of WHO, COVID-19 outbreak considered as a public health problem of international concern. The aim of this study is to provide a comprehensive view on COVID-19 and the future expectations to control virus progression. Patients with liver disease, diabetes, high blood pressure, and obesity are more susceptible to the incidence of COVID-19 infection. So, there is a rapid need for disease diagnosis, vaccine development, and drug discovery to detect, prevent, and treat this sudden and lethal virus. Real-time polymerase chain reaction (RT-PCR) is considered as a rapid, accurate, and specific tool for disease diagnosis. Under this emergency situation that the world facing against COVID-19, there are about 15 potential vaccine candidates tested globally based on messenger RNA, DNA-based, nanoparticle, synthetic, and modified virus-like particle. Certain drugs that are clinically approved for other diseases were tested against COVID-19 as chloroquine, hydroxychloroquine, ivermectin, favipiravir, ribavirin, and remdesivir. Convalescent plasma transfusion and traditional herbal medicine were also taken into consideration. Due to the absence of effective treatment or vaccines against COVID-19 so far, the precautionary measures according to WHO’s strategic objectives are the only way to confront this crisis. Governments should adopt national medical care programs to reduce the risk of exposure to any future viral outbreaks especially to patients with pre-existing medical conditions.


title: Effects of Additional Milk Replacer Feeding on Calf Health, Growth, and Selected Blood Metabolites in Calves

text: Abstract The objective of the experiment was to evaluate effects of increased milk replacer feeding on growth, intake, feed efficiency, and health parameters in stressed calves. Holstein bull calves (n = 120; approximately 3 to 8 d of age) were purchased from sale barns and dairy farms and housed in fiberglass hutches. In addition, wood shavings contaminated with coronavirus were mixed with clean shavings and added to each hutch before the start of the experiment. Calves were fed either a fixed amount (454 g/d) of a 20% crude protein (CP), 20% fat milk replacer to weaning at 28 d or a variable amount (454, 681, 908, and 454 g/d on d 0 to 7, 8 to 14, 15 to 31, and 32 to 41, respectively) of a milk replacer containing 28% CP and 17% fat without or with added dietary supplement containing bovine serum. Calves were also fed commercial calf starter and water ad libitum. Plasma IgG concentration in most calves on arrival at the facility was<10g/L. Intake, change in body weight, feed efficiency, morbidity and mortality, and selected plasma metabolites were determined. Body weight at 28 d, 56 d, daily body weight gain, intake of milk replacer, fecal scores, days with diarrhea, and days treated with antibiotics were increased with feeding variable amount of milk replacer over the 56-d study. Starter intake from d 1 to 56 was reduced from 919 to 717 g/d in calves fed fixed and variable amounts of milk replacer, respectively. Morbidity, measured as the number of days that calves had diarrhea, was increased by 53% when a variable amount of milk replacer was fed. Calves fed variable milk replacer were treated with antibiotics for 3.1 d compared with 1.9 d for calves fed 454g of milk replacer/d. Concentrations of plasma glucose, urea N, and insulin-like growth factor-I were increased when calves were fed variable amount of milk replacer. Dietary supplement containing bovine serum had no effect on any parameter measured. There was no effect of milk replacer feeding on concentrations of nonesterified fatty acids, total protein, or growth hormone concentrations. Plasma tumor necrosis factor-α was highest in calves with the highest plasma IgG concentrations on the day of arrival and might be related to the calf's ability to identify pathogens in the environment. Under conditions of this study, calves fed variable amount of milk replacer and exposed to immunological challenge before weaning had greater BW gain, but also increased incidence of diarrhea that required added veterinary treatments.


title: Dieckol, a SARS-CoV 3CLpro inhibitor, isolated from the edible brown algae Ecklonia cava

text: Phlorotannins have been isolated and are the first of their type showing competitively inhibitory activity toward SARS-CoV 3CLpro. Dieckol showed the most potent SARS-CoV 3CLpro trans/cis-cleavage inhibitory effects with high association rate. [Figure: see text]


COVID‐19 and childhood acute lymphoblastic leukemia


A One-Health lens for anthrax


title: The ubiquitin-proteasome system is required for the early stages of porcine circovirus type 2 replication

text: Abstract Porcine circovirus type 2 (PCV2) is the primary causative agent of porcine circovirus-associated diseases (PCVAD). It has been shown that the ubiquitin-proteasome system (UPS) is correlated with viral infection, but its role in PCV2 replication remains unknown. In the present study, we explored the interplay between PCV2 replication and the UPS in PK15 cells and found that treatment with a proteasome inhibitor (MG132 and lactacystin) significantly decreased the PCV2 titer at the early infection stage. We further revealed that inhibition of the UPS did not affect virus entry but decreased viral protein expression and RNA transcription potentially in a cell cycle-dependent manner. PCV2 infection has little effect on the chymotrypsin-like activity, and the gene-silencing of ubiquitin reduced the PCV2 titer, which indicates that the effective replication of PCV2 may be related to protein ubiquitination. Taken together, our data suggested that PCV2 replication requires the UPS machinery, which may represent a potential antiviral target against PCV2.


title: Short Term Effects of Weather on Hand, Foot and Mouth Disease

text: BACKGROUND: Hand, foot, and mouth disease (HFMD) outbreaks leading to clinical and fatal complications have increased since late 1990s; especially in the Asia Pacific Region. Outbreaks of HFMD peaks in the warmer season of the year, but the underlying factors for this annual pattern and the reasons to the recent upsurge trend have not yet been established. This study analyzed the effect of short-term changes in weather on the incidence of HFMD in Singapore. METHODS: The relative risks between weekly HFMD cases and temperature and rainfall were estimated for the period 2001–2008 using time series Poisson regression models allowing for over-dispersion. Smoothing was used to allow non-linear relationship between weather and weekly HFMD cases, and to adjust for seasonality and long-term time trend. Additionally, autocorrelation was controlled and weather was allowed to have a lagged effect on HFMD incidence up to 2 weeks. RESULTS: Weekly temperature and rainfall showed statistically significant association with HFMD incidence at time lag of 1–2 weeks. Every 1°C increases in maximum temperature above 32°C elevated the risk of HFMD incidence by 36% (95% CI = 1.341–1.389). Simultaneously, one mm increase of weekly cumulative rainfall below 75 mm increased the risk of HFMD by 0.3% (CI = 1.002–1.003). While above 75 mm the effect was opposite and each mm increases of rainfall decreased the incidence by 0.5% (CI = 0.995–0.996). We also found that a difference between minimum and maximum temperature greater than 7°C elevated the risk of HFMD by 41% (CI = 1.388–1.439). CONCLUSION: Our findings suggest a strong association between HFMD and weather. However, the exact reason for the association is yet to be studied. Information on maximum temperature above 32°C and moderate rainfall precede HFMD incidence could help to control and curb the up-surging trend of HFMD.


title: Umgang mit COVID-19 in der Notaufnahme: Erfahrungsbericht der interdisziplinären Notaufnahme des Universitätsklinikums Münster

text: With the COVID-19 pandemic, emergency rooms are faced with major challenges because they act as the interface between outpatient and inpatient care. The dynamics of the pandemic forced emergency care at the University Hospital Münster to extensively adjust their processes, which had to be carried out in the shortest time possible. This included the establishment of an outpatient coronavirus test center and a medical student-operated telephone hotline. Inside the hospital, new isolation capacities in the emergency room and a dedicated COVID-19 ward were set up. The patient flow was reorganized using flow diagrams for both the outpatient and inpatient areas. The general and special emergency management was optimized for the efficient treatment of COVID-19-positive patients and the staff were trained in the use of protective equipment. This report of our experience is intended to support other emergency departments in their preparation for the COVID-19 pandemic.


Severe acute respiratory syndrome spreads worldwide


title: Molecular characterization of the 9.36 kb C-terminal region of canine coronavirus 1-71 strain

text: A total of 9.36 kb nucleotides of the C-terminal one-third genome of the canine Coronavirus (CCV) 1-71 strain, including all the structural protein genes and some non-structural protein (nsp) genes were cloned and sequenced. Nucleotide and amino acid sequence alignment as well as phylogenetic analysis of all the structural ORFs with reference coronavirus strains showed that CCV 1-71 was highly related to Chinese CCV isolates. This indicates that these CCV stains may have the same ancestor. Two large deletions were found in ORF3 and led to an elongated nsp3a and a truncated nsp3b. A single “A” insertion in a 6A stretch resulted in the truncation of nsp7b. The great variation in nsp3b and nsp7b indicates that these proteins are not essential for the viral replication.


title: Positive effects of COVID-19 control measures on pneumonia prevention

text: • pneumonia can be stopped while fight against the SARS-COV-2 outbreak; • pneumonia cases (except COVID-19 cases) were dramatically decreased since the control measures were onset; • Other respiratory infection diseases may also to some extent be stopped while fighting against the pandemic of COVID-19.


title: Coexistence of Covid-19 and Acute Ischemic Stroke Report of Four Cases

text: Abstract Coronaviruses are revealed to target the human respiratory system mainly. However, they also have neuro-invasive abilities and might spread from the respiratory system to the central nervous system. Herein, we report four patients with COVID-19 simultaneously diagnosed with acute ischemic stroke. There were four stroke cases with simultaneously diagnosis of Covid-19 till the April 14, 2020 in the city of Sakarya, Turkey. They were aged between 45 to 77 years. All four cases were likely to have contracted the virus in Sakarya. The patients had all commonly reported symptoms of Covid-19. Three patients have elevated D-dimer levels, and two of them had high C-reactive protein (CRP) levels. They were managed symptomatically for both the infection and the stroke. Our findings suggest that ischemic cerebrovascular diseases may simultaneously develop in the course of Covid-19 independently of the critical disease process. Increased inflammation predicted by CRP and D-dimer levels may play a role in the formation of ischemia. In particular, elder patients with prothrombotic risk factors should also be considered for the signs of cerebrovascular events in addition to infectious symptoms.


COVID-19 in Nursing Homes


Retos clínicos en pacientes con enfermedad pulmonar obstructiva crónica que han sufrido una infección por SARS-CoV-2


title: A Review of Models Used for Investigating Barriers to Healthcare Access in Australia

text: Understanding barriers to healthcare access is a multifaceted challenge, which is often highly diverse depending on location and the prevalent surroundings. The barriers can range from transport accessibility to socio-economic conditions, ethnicity and various patient characteristics. Australia has one of the best healthcare systems in the world; however, there are several concerns surrounding its accessibility, primarily due to the vast geographical area it encompasses. This review study is an attempt to understand the various modeling approaches used by researchers to analyze diverse barriers related to specific disease types and the various areal distributions in the country. In terms of barriers, the most affected people are those living in rural and remote parts, and the situation is even worse for indigenous people. These models have mostly focused on the use of statistical models and spatial modeling. The review reveals that most of the focus has been on cancer-related studies and understanding accessibility among the rural and urban population. Future work should focus on further categorizing the population based on indigeneity, migration status and the use of advanced computational models. This article should not be considered an exhaustive review of every aspect as each section deserves a separate review of its own. However, it highlights all the key points, covered under several facets which can be used by researchers and policymakers to understand the current limitations and the steps that need to be taken to improve health accessibility.


title: It’s a small world after all: A Canadian resident’s perspective on COVID-19

text: COVID-19 has infected millions of people, with an estimated total dead in the hundreds of thousands. This has significantly impacted health care, including who is delivering it, how it is delivered, and how it is taught. This article describes challenges of the COVID-19 pandemic from the perspective of a Canadian nuclear medicine resident, including new risks with nuclear imaging, navigating new and sometimes challenging guidelines, as well as working and living within the confines of social distancing.


title: Hyper Acute Demyelinating Encephalomyelitis of Childhood: A Rare Entity

text: A young child with catastrophic neurological illness diagnosed as a rare variant of acute demyelinating encephalomyelitis (ADEM). She succumbed to her illness despite of aggressive and appropriate management. Malignant demyelinating encephalomyelitis should be considered in children who are refractory to the treatment of ADEM.


title: A killed Leishmania vaccine with sand fly saliva extract and saponin adjuvant displays immunogenicity in dogs

text: A vaccine against canine visceral leishmaniasis (CVL), comprising Leishmania braziliensis promastigote protein, sand fly gland extract (SGE) and saponin adjuvant, was evaluated in dog model, in order to analyse the immunogenicity of the candidate vaccine. The vaccine candidate elicited strong antigenicity in dogs in respect of specific SGE and Leishmania humoral immune response. The major saliva proteins recognized by serum from immunized dogs exhibited molecular weights of 35 and 45 kDa, and were related to the resistance pattern against Leishmania infection. Immunophenotypic analysis revealed increased circulating CD21(+) B-cells and CD5(+) T-cells, reflected by higher counts of CD4(+) and CD8(+) T-cells. The observed interaction between potential antigen-presenting cells (evaluated as CD14(+) monocytes) and lymphocyte activation status indicated a relationship between innate and adaptive immune responses. The higher frequency in L. chagasi antigen-specific CD8(+) T-lymphocytes, and their positive association with intense cell proliferation, in addition to the progressively higher production of serum nitric oxide levels, showed a profile compatible with anti-CVL vaccine potential. Further studies on immunological response after challenge with L. chagasi may provide important information that will lead to a better understanding on vaccine trial and efficacy.


title: 5 Basing a Nonclinician's Career upon Simulation The Personal Experience of a Physicist

text: Publisher Summary This chapter concerns the breed of people who start by selecting a nonclinician path and eventually work in clinical simulation. Their new role requires their grasping a wide range of clinical and educational concepts and skills that should already be second nature to their new colleagues who are trained as an educator, a clinician, or health care professional. There are many different opportunities and responsibilities in the domain of clinical simulation, and nonclinicians could occupy almost any one of them. This ranges from educationalist, communication or team dynamic expert, technician, or technical manager, center coordinator, administrator, to operations manager. The background of potential simulation center employees will be an important asset in their new role as it could give them a different perspective, additional skills, and innovative ideas that complement those of their clinical colleagues.


title: An ultrasensitive, rapid, and portable coronavirus SARS-CoV-2 sequence detection method based on CRISPR-Cas12

text: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has received global attention due to the recent outbreak in China. In this work, we report a CRISPR-Cas12 based diagnostic tool to detect synthetic SARS-CoV-2 RNA sequences in a proof-of-principle evaluation. The test proved to be sensitive, rapid, and potentially portable. These key traits of the CRISPR method are critical for virus detection in regions that lack resources to use the currently available methods.


title: Epidémiologie virale et sévérité clinique pendant le pic d’épidémie grippale A(H1N1) variant dans les atteintes respiratoires fébriles de l’enfant

text: Résumé En mars 2009, un nouveau virus a fait émergence, le virus Influenzae A(H1N1) variant. Nous avons voulu en étudier la répercussion médicale. Objectifs Définir la prévalence virale dans les atteintes respiratoires fébriles hospitalisées de l’enfant en période d’épidémie grippale A(H1N1) variant et déterminer les caractéristiques cliniques, paracliniques et évolutives liées aux virus identifiés. Méthodes Il s’est agi d’une étude épidémiologique, prospective, monocentrique menée à l’hôpital des Enfants du centre hospitalier universitaire de Bordeaux (Aquitaine, France) pendant l’épidémie grippale du 23 novembre au 20 décembre 2009 inclus. Tous les enfants hospitalisés pour atteinte respiratoire fébrile ont été inclus. Résultats Soixante-treize enfants ont été inclus. Une identification virale a été possible dans 52 % des cas, incluant 23 % de A(H1N1) variant [A(H1N1)v] et 29 % d’autres identifications dont 22 % de virus respiratoires syncitiaux. Une seule co-infection a été observée entre le virus [A(H1N1)v] et un pool viral (adénovirus ou virus para-influenzae ou bocavirus). Il n’existait pas de différence significative concernant l’âge, le sexe ou les facteurs de risque de gravité en fonction des virus identifiés. Concernant le virus [A(H1N1)v], les symptômes les plus fréquents étaient l’altération de l’état général, la toux, les atteintes oto-rhino-laryngologiques et les anomalies de la fréquence respiratoire, avec significativement moins de signes de lutte et d’anomalie auscultatoire mais plus de convulsions. Sur le plan paraclinique, il n’y avait pas de différence significative entre les groupes. Évolution et prise en charge ont été comparables. Conclusion Une faible prédominance du virus [A(H1N1)v] a été notée pendant l’épidémie grippale en Aquitaine dans les atteintes respiratoires fébriles justifiant une hospitalisation. Les caractéristiques cliniques, paracliniques et évolutives étaient peu spécifiques. L’ensemble de ces éléments n’a pas révélé de gravité particulière liée au virus [A(H1N1)v]. Summary In 2009, a new emerging flu virus, A(H1N1), was identified. Its true medical impact on children's health remains widely debated. Aim To define the prevalence of respiratory disease in children hospitalized with fever during the influenza A(H1N1) epidemic and to determine the clinical, paraclinical, and outcome characteristics according to the viruses identified. Material and methods Children hospitalized for a febrile respiratory disease were included in this prospective cohort study conducted at Bordeaux University's Children's Hospital (France) during the influenza epidemic from 2009/11/23 to 2009/12/20. Results Seventy-three children were included in the study. Viruses were identified by PCR in 52% (38/73) of cases, including 23% (17/73) A(H1N1) virus and 29% (21/73) other viruses, 22% (16/73) of which were syncytial respiratory viruses. There was only one case of co-infection between A(H1N1) virus and another virus from the para-influenza virus or adenovirus or bocavirus pool. No significant difference regarding age, sex, or risk factors in the different viral groups was noted. Regarding the A(H1N1) virus, the most frequent symptoms were deterioration of the overall health status, cough, ENT disease, and rapid breathing, with significantly less increased breathing effort and auscultatory abnormality albeit with more seizures. There was no significant difference between groups regarding laboratory data. Management and outcome were similar. Conclusion The prevalence of A(H1N1) virus during the 2009 epidemic in Aquitaine was low among febrile hospitalized children with breathing symptoms. Clinical and paraclinical signs were non-specific. The tolerance and prognosis of influenza A(H1N1) infection in children was satisfactory.


title: Opioid Policy Changes During the COVID-19 Pandemic - and Beyond

text: The United States is currently in the midst of 2 public health emergencies: COVID-19 and the ongoing opioid crisis. In an attempt to reduce preventable harm to individuals with opioid use disorder (OUD), federal, state, and local governments have temporarily modified law and policy to increase access to OUD treatment and divert some individuals at high risk away from the correctional system. In this Commentary, we briefly describe how people with OUD are at increased risk for COVID-19, discuss existing policy barriers to evidence-based prevention and treatment for individuals with OUD, explain the temporary rollbacks of those barriers, and argue that these changes should be made permanent. We also suggest several additional steps that federal and state governments can urgently take to reduce barriers to care for individuals with OUD, both during the current crisis and beyond.


Type of Manuscript: Letter to the EditorsSyndrome of Inappropriate Antidiuretic Hormone as the Initial Presentation of COVID-19: A Novel Case Report


title: Potential antivirals and antiviral strategies against SARS coronavirus infections

text: There are a number of antivirals as well as antiviral strategies that could be envisaged to prevent or treat severe acute respiratory syndrome (SARS) (or similar) coronavirus (CoV) infections. Targets for the prophylactic or therapeutic interventions include interaction of the spike (S) glycoprotein (S1 domain) with the host cell receptor, fusion of the S2 domain with the host cell membrane, processing of the replicase polyproteins by the virus-encoded proteases (3C-like cysteine protease [3CLpro] and papain-like cysteine protease) and other virus-encoded enzymes such as the NTPase/helicase and RNA-dependent RNA polymerase. Human monoclonal antibody blocking S1 may play an important role in the immunoprophylaxis of SARS. Fusion inhibitors reminiscent of enfuvirtide in the case of HIV may also be developed for SARS-CoV. Various peptidomimetic and nonpeptidic inhibitors of 3CLpro have been described, the best ones inhibiting SARS-CoV replication with a selectivity index greater than 1000. Human interferons, in particular α- and β-interferon, as well as short interfering RNAs could further be pursued for the control of SARS. Various other compounds, often with an ill-defined mode of action but selectivity indexes up to 100, have been reported to exhibit in vitro activity against SARS-CoV: valinomycin, glycopeptide antibiotics, plant lectins, hesperetin, glycyrrhizin, aurintricarboxylic acid, chloroquine, niclosamide, nelfinavir and calpain inhibitors.


title: Host Subtraction, Filtering and Assembly Validations for Novel Viral Discovery Using Next Generation Sequencing Data

text: The use of next generation sequencing (NGS) to identify novel viral sequences from eukaryotic tissue samples is challenging. Issues can include the low proportion and copy number of viral reads and the high number of contigs (post-assembly), making subsequent viral analysis difficult. Comparison of assembly algorithms with pre-assembly host-mapping subtraction using a short-read mapping tool, a k-mer frequency based filter and a low complexity filter, has been validated for viral discovery with Illumina data derived from naturally infected liver tissue and simulated data. Assembled contig numbers were significantly reduced (up to 99.97%) by the application of these pre-assembly filtering methods. This approach provides a validated method for maximizing viral contig size as well as reducing the total number of assembled contigs that require down-stream analysis as putative viral nucleic acids.


Chronic Use of Angiotensin-Converting Enzyme Inhibitors and Angiotensin II Receptor Blockers Is High Among Intensive Care Unit Patients With Non–COVID-19 Sepsis but Carries a Moderately Increased Risk of Death


title: Chapter 7 Indoor Environmental Quality

text: Publisher Summary The chapter discusses the indoor environmental quality. A variety of factors have been found to contribute to poor indoor-air quality in buildings, the primary one being indoor pollution sources that release gases or particles into the air and the other major sources include outdoor pollutants . The chapter illustrates the factor affecting the indoor environmental quality which are inorganic contaminants, contaminants generated by combustion, organic contaminants, biological contaminants, and reducing exposure. People from different cultures have different comfort zones; even people belonging to the same family may feel comfortable under different conditions, and keeping everyone comfortable at the same time is an elusive goal at best. Thermal comfort involves temperature, humidity, and air movement. A comfort zone includes an optimal range and combinations of thermal factors, in which at least 80% of the building occupants feel satisfied. Ventilation is vital for the health and comfort of building occupants. It is needed to reduce and remove pollutants emitted from various internal and external sources. Good design combined with optimum air tightness is prerequisite to ensure healthy air quality, occupant comfort, and energy efficiency. There has been a lack of systematic experiments in which building materials are ranked according to their pollution strength and the impact on the indoor-air quality of using these materials in real rooms analyzed. These studies allow to quantify the extent to which using low-polluting building materials would reduce the energy needed for ventilation of buildings without compromising indoor-air quality.


title: Reducing Poverty among Children: Evidence from State Policy Simulations

text: Abstract State approaches to reducing child poverty vary considerably. We exploit this state-level variation to estimate what could be achieved in terms of child poverty if all states adopted the most generous or inclusive states’ policies. Specifically, we simulate the child poverty reductions that would occur if every state were as generous or inclusive as the most generous or inclusive state in four key policies: Supplemental Nutrition Assistance Program (SNAP), state Earned Income Tax Credits (EITC), Temporary Assistance for Needy Families (TANF), and state Child Tax Credits (CTC). We find that adopting the most generous or inclusive state EITC policy would have the largest impact on child poverty, reducing it by 1.2 percentage points, followed by SNAP, TANF, and lastly state CTC. If all states were as generous or inclusive as the most generous or inclusive state in all four policies, the child poverty rate would decrease by 2.5 percentage points, and five and a half million children would be lifted out of poverty.


title: Accelerated FoxP2 Evolution in Echolocating Bats

text: FOXP2 is a transcription factor implicated in the development and neural control of orofacial coordination, particularly with respect to vocalisation. Observations that orthologues show almost no variation across vertebrates yet differ by two amino acids between humans and chimpanzees have led to speculation that recent evolutionary changes might relate to the emergence of language. Echolocating bats face especially challenging sensorimotor demands, using vocal signals for orientation and often for prey capture. To determine whether mutations in the FoxP2 gene could be associated with echolocation, we sequenced FoxP2 from echolocating and non-echolocating bats as well as a range of other mammal species. We found that contrary to previous reports, FoxP2 is not highly conserved across all nonhuman mammals but is extremely diverse in echolocating bats. We detected divergent selection (a change in selective pressure) at FoxP2 between bats with contrasting sonar systems, suggesting the intriguing possibility of a role for FoxP2 in the evolution and development of echolocation. We speculate that observed accelerated evolution of FoxP2 in bats supports a previously proposed function in sensorimotor coordination.


Beyond the Routine Influenza Surveillance


The challenge of breath: toward an ‘after’ COVID‐19


Don’t throw the stethoscope away!: The value of the stethoscope over ultrasound equipment during the COVID-19 outbreak is discussed by authors from China


title: Bilateral posterior infarction in a SARS-Cov-2 infected patient: discussion about an unusual case

text: In time of SARS-Cov2 pandemic, neurologists need to be vigilant for cerebrovascular complications of Covid-19. We present a case of bilateral occipito-temporal infarction revealed by a sudden cortical blindness with haemorrhagic transformation after intravenous thrombolysis in a diabetic patient infected by Covid-19. Differential diagnoses are discussed in front of this unusual presentation and evolution.


The Role of Transthoracic Ultrasound in the novel Coronavirus Disease (COVID-19): A Reappraisal. Information and Disinformation: Is There Still Place for a Scientific Debate?


title: Identification of a contemporary human parechovirus type 1 by VIDISCA and characterisation of its full genome

text: BACKGROUND: Enteritis is caused by a spectrum of viruses that is most likely not fully characterised. When testing stool samples by cell culture, virus isolates are sometimes obtained which cannot be typed by current methods. In this study we used VIDISCA, a virus identification method which has not yet been widely applied, on such an untyped virus isolate. RESULTS: We found a human parechovirus (HPeV) type 1 (strain designation: BNI-788st). Because genomes of contemporary HPeV1 were not available, we determined its complete genome sequence. We found that the novel strain was likely the result of recombination between structural protein genes of an ancestor of contemporary HPeV1 strains and nonstructural protein genes from an unknown ancestor, most closely related to HPeV3. In contrast to the non-structural protein genes of other HPeV prototype strains, the non-structural protein genes of BNI-788st and HPeV3 prototype strains did not co-segregate in bootscan analysis with that of other prototype strains. CONCLUSION: HPeV3 nonstructural protein genes may form a distinct element in a pool of circulating HPeV non-structural protein genes. More research into the complex HPeV evolution is required to connect virus ecology with disease patterns in humans.


title: Vaxign: The First Web-Based Vaccine Design Program for Reverse Vaccinology and Applications for Vaccine Development

text: Vaxign is the first web-based vaccine design system that predicts vaccine targets based on genome sequences using the strategy of reverse vaccinology. Predicted features in the Vaxign pipeline include protein subcellular location, transmembrane helices, adhesin probability, conservation to human and/or mouse proteins, sequence exclusion from genome(s) of nonpathogenic strain(s), and epitope binding to MHC class I and class II. The precomputed Vaxign database contains prediction of vaccine targets for >70 genomes. Vaxign also performs dynamic vaccine target prediction based on input sequences. To demonstrate the utility of this program, the vaccine candidates against uropathogenic Escherichia coli (UPEC) were predicted using Vaxign and compared with various experimental studies. Our results indicate that Vaxign is an accurate and efficient vaccine design program.


title: Role of vascular cell adhesion molecules and leukocyte apoptosis in the lymphopenia and thrombocytopenia of patients with severe acute respiratory syndrome (SARS)

text: Abstract The immunopathogenesis of leukopenia and thrombocytopenia in patients with severe acute respiratory syndrome (SARS) is unclear. In order to explore the leukopenia mechanism, we studied 15 SARS patients who were previously healthy, and 15 age-matched normal controls in a paired design. Soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble Fas ligand (sFasL) in plasma were measured by ELISA, and intracellular activated caspase-3 fragment in different leukocytes was determined by flow cytometry. Patients with SARS had significantly lower lymphocyte and platelet counts and significantly higher sVCAM-1 and sFasL levels compared to healthy controls. sVCAM-1 levels correlated negatively with total leukocytes and platelet counts, but positively with plasma sFasL levels. Intracellular cleaved caspase-3 expression was also significantly higher in lymphocytes from SARS patients in acute phase than in convalescent stage. Lymphopenia and thrombocytopenia in SARS patients may be caused, in part, by enhanced vascular sequestration associated with increased sVCAM-1 levels. However, lymphopenia may be due to enhanced cell death. Inhibition of cell adhesion and caspase-3 activation could, therefore, have prevented SARS patients from developing thrombocytopenia and lymphopenia.


title: Discovery of unsymmetrical aromatic disulfides as novel inhibitors of SARS-CoV main protease: Chemical synthesis, biological evaluation, molecular docking and 3D-QSAR study

text: The worldwide outbreak of severe acute respiratory syndrome (SARS) in 2003 had caused a high rate of mortality. Main protease (M(pro)) of SARS-associated coronavirus (SARS-CoV) is an important target to discover pharmaceutical compounds for the therapy of this life-threatening disease. During the course of screening new anti-SARS agents, we have identified that a series of unsymmetrical aromatic disulfides inhibited SARS-CoV M(pro) significantly for the first time. Herein, 40 novel unsymmetrical aromatic disulfides were synthesized chemically and their biological activities were evaluated in vitro against SARS-CoV M(pro). These novel compounds displayed excellent IC(50) data in the range of 0.516–5.954 μM. Preliminary studies indicated that these disulfides are reversible and mpetitive inhibitors. A possible binding mode was generated via molecular docking simulation and a comparative field analysis (CoMFA) model was constructed to understand the structure-activity relationships. The present research therefore has provided some meaningful guidance to design and identify anti-SARS drugs with totally new chemical structures.


title: Inhibition of cytokine signaling by ruxolitinib and implications for COVID-19 treatment

text: Approximately 15% of patients with coronavirus disease 2019 (COVID-19) experience severe disease, and 5% progress to critical stage that can result in rapid death. No vaccines or antiviral treatments have yet proven effective against COVID-19. Patients with severe COVID-19 experience elevated plasma levels of pro-inflammatory cytokines, which can result in cytokine storm, followed by massive immune cell infiltration into the lungs leading to alveolar damage, decreased lung function, and rapid progression to death. As many of the elevated cytokines signal through Janus kinase (JAK)1/JAK2, inhibition of these pathways with ruxolitinib has the potential to mitigate the COVID-19–associated cytokine storm and reduce mortality. This is supported by preclinical and clinical data from other diseases with hyperinflammatory states, where ruxolitinib has been shown to reduce cytokine levels and improve outcomes. The urgent need for treatments for patients with severe disease support expedited investigation of ruxolitinib for patients with COVID-19.


title: RNA sequence and secondary structural determinants in a minimal viral promoter that directs replicase recognition and initiation of genomic plus-strand RNA synthesis 1 1 Edited by D. E. Draper

text: Abstract Viral RNA replication provides a useful system to study the structure and function of RNAs and the mechanism of RNA synthesis from RNA templates. Previously we demonstrated that a 27 nt RNA from brome mosaic virus (BMV) can direct correct initiation of genomic plus-strand RNA synthesis by the BMV replicase. In this study, using biochemical, nuclear magnetic resonance, and thermodynamic analyses, we determined that the secondary structure of this 27 nt RNA can be significantly altered and retain the ability to direct RNA synthesis. In contrast, we find that position-specific changes in the RNA sequence will affect replicase recognition, modulate the polymerization process, and contribute to the differential accumulation of viral RNAs. These functional results are in agreement with the phylogenetic analysis of BMV and related viral sequences and suggest that a similar mechanism of RNA synthesis takes place for members of the alphavirus superfamily.


title: Preparation of Cultured Cells Using High-Pressure Freezing and Freeze Substitution for Subsequent 2D or 3D Visualization in the Transmission Electron Microscope

text: Transmission electron microscopy (TEM) is an invaluable technique used for imaging the ultrastructure of samples and it is particularly useful when determining virus–host interactions at a cellular level. The environment inside a TEM is not favorable for biological material (high vacuum and high energy electrons). Also biological samples have little or no intrinsic electron contrast, and rarely do they naturally exist in very thin sheets, as is required for optimum resolution in the TEM. To prepare these samples for imaging in the TEM therefore requires extensive processing which can alter the ultrastructure of the material. Here we describe a method which aims to minimize preparation artifacts by freezing the samples at high pressure to instantaneously preserve ultrastructural detail, then rapidly substituting the ice and infiltrating with resin to provide a firm matrix which can be cut into thin sections for imaging. Thicker sections of this material can also be imaged and reconstructed into 3D volumes using electron tomography.


title: Clinical and CT features of the COVID-19 infection: comparison among four different age groups

text: PURPOSE: To compare and analyze the clinical and CT features of coronavirus disease 2019 (COVID-19) among four different age groups. METHODS: 97 patients (45 males, 52 females, mean age, 66.2 ± 5.0) with chest CT examination and positive reverse transcriptase-polymerase chain reaction test (RT-PCR) from January 17, 2020 to February 21, 2020 were retrospectively studied. The patients were divided into four age groups (children [0–17 years], young adults [18–44 years], middle age [45–59 years], and senior [≥ 60 years]) according to their age after the diagnosis was made based on PCR test and clinical symptoms. RESULTS: Comorbidities such as hypertension, diabetes mellitus, and heart disease are more common in the senior group. Cluster onset (two or more confirmed cases in a small area) is more common in the children group and senior group. Older patients were found to have a higher incidence of the highest clinical classification (severe or critical) in these four groups. Senior patients have a higher incidence of large/multiple ground-glass opacity (GGO). Child patients are mostly negative for chest CT or with involvement of only one lobe of the lung; while in older patients, there was a higher incidence of involvement of four or five lung lobes. The frequency of lobe involvement was also found to have significant differences in the four age groups. CONCLUSION: The clinical and imaging features of patients in different age groups were found to be significantly different. A better understanding of the age differences in comorbidities, cluster onset, highest clinical classification, large/multiple GGO, numbers of lobes affected, and frequency of lobe involvement can be useful in the diagnosis of COVID-19 patients of different ages.


title: Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro

text: Transmissible gastroenteritis virus (TGEV) infection causes severe diarrhea in piglets and imposes a significant economic burden on pig farms. Single-chain fragment variable (scFv) antibodies effectively inhibit virus infection and could be a potential therapeutic reagent for preventing disease. In this study, a recombinant scFv antibody phage display library was constructed from peripheral blood lymphocytes of piglets infected with TGEV. The library was screened with four rounds of biopanning using purified TGEV antigen, and scFv antibodies that bound to TGEV were obtained. The scFv gene was subcloned into the pET-28a(+), and the constituted plasmid was introduced into Escherichia coli BL21 (DE3) for protein expression. All three scFv clones identified had neutralizing activity against TGEV. An immunofluorescence assay and western blot analysis demonstrated that two scFv antibodies reacted with the spike protein of TGEV. These results indicate that scFv antibodies provide protection against viral infection in vitro and may be a therapeutic candidate for both prevention and treatment of TGEV infection in swine.


title: Microbiological testing of adults hospitalised with community-acquired pneumonia: an international study

text: This study aimed to describe real-life microbiological testing of adults hospitalised with community-acquired pneumonia (CAP) and to assess concordance with the 2007 Infectious Diseases Society of America (IDSA)/American Thoracic Society (ATS) and 2011 European Respiratory Society (ERS) CAP guidelines. This was a cohort study based on the Global Initiative for Methicillin-resistant Staphylococcus aureus Pneumonia (GLIMP) database, which contains point-prevalence data on adults hospitalised with CAP across 54 countries during 2015. In total, 3702 patients were included. Testing was performed in 3217 patients, and included blood culture (71.1%), sputum culture (61.8%), Legionella urinary antigen test (30.1%), pneumococcal urinary antigen test (30.0%), viral testing (14.9%), acute-phase serology (8.8%), bronchoalveolar lavage culture (8.4%) and pleural fluid culture (3.2%). A pathogen was detected in 1173 (36.5%) patients. Testing attitudes varied significantly according to geography and disease severity. Testing was concordant with IDSA/ATS and ERS guidelines in 16.7% and 23.9% of patients, respectively. IDSA/ATS concordance was higher in Europe than in North America (21.5% versus 9.8%; p<0.01), while ERS concordance was higher in North America than in Europe (33.5% versus 19.5%; p<0.01). Testing practices of adults hospitalised with CAP varied significantly by geography and disease severity. There was a wide discordance between real-life testing practices and IDSA/ATS/ERS guideline recommendations.


title: Middle East respiratory syndrome coronavirus in Al-Madinah City, Saudi Arabia: Demographic, clinical and survival data

text: Abstract Background Middle East respiratory syndrome coronavirus (MERS-CoV), is an emerging virus respiratory infection. It has a high mortality rate and a wide spectrum of clinical features. This study describes the clinical characteristics and outcome of MERS infected patients. Methods A retrospective study was conducted of all confirmed MERS-CoV infections from March 2014 to May 2014 at two tertiary care hospitals in Al-Madinah region (Saudi Arabia). We gathered data about demographic, clinical presentation, and factors associated with severity and mortality. Results A total of 29 cases were identified; 20 males (69%) and nine females (31%), age 45±12years. The death rate was higher for men (52%) than for women (23%). Initial presentation was fever in 22 (75%) cases, cough in 20 (69%) cases, and shortness of breath in 20 (69%) cases. Associated comorbidities were diabetes mellitus in nine (31%) patients and chronic kidney disease (CKD) in eight (27%) patients. Duration of symptoms before hospitalization ranged from 2.9days to 5days. Elevated liver enzymes were present in 14 (50%) patients and impaired renal profile present in eight (27%) patients. We also describe in this study radiological patterns and factors associated with mortality. Conclusion MERS-CoV infection transmission continues to occur as clusters in healthcare facilities. The frequency of cases and deaths is higher among men than women and among patients with comorbidities.