title: Living with the enemy or uninvited guests: Functional genomics approaches to investigating host resistance or tolerance traits to a protozoan parasite, Theileria annulata, in cattle

text: Abstract Many breeds of cattle with long histories of living in areas of endemic disease have evolved mechanisms that enable them to co-exist with specific pathogens. Understanding the genes that control tolerance and resistance could provide new strategies to improve the health and welfare of livestock. Around one sixth of the world cattle population is estimated to be at risk from one of the most debilitating tick-borne diseases of cattle, caused by the protozoan parasite, Theileria annulata. The parasite mainly infects cells of the myeloid lineage which are also the main producers of inflammatory cytokines. If an infectious or inflammatory insult is sufficiently great, inflammatory cytokines produced by macrophages enter the circulation and induce an acute phase proteins (APP) response. The Bos taurus Holstein breed produces higher and more prolonged levels of inflammatory cytokine induced APP than the Bos indicus Sahiwal breed in response to experimental infection with T. annulata. The Sahiwal exhibits significantly less pathology and survives infection, unlike the Holstein breed. Therefore, we hypothesised that the causal genes were likely to be expressed in macrophages and control the production of inflammatory cytokines. A functional genomics approach revealed that the transcriptome profile of the B. taurus macrophages was more associated with an inflammatory programme than the B. indicus macrophages. In particular the most differentially expressed gene was a member of the signal regulatory protein (SIRP) family. These are mainly expressed on myeloid cell surfaces and control inflammatory responses. Other differentially expressed genes included bovine major histocompatibility complex (MHC) (BoLA) class II genes, particularly BoLA DQ, and transforming growth factor (TGF)B2. We are now exploring whether sequence and functional differences in the bovine SIRP family may underlie the resistance or tolerance to T. annulata between the breeds. Potentially, our research may also have more general implications for the control of inflammatory processes against other pathogens. Genes controlling the balance between pathology and protection may determine how livestock can survive in the face of infectious onslaught. Next generation sequencing and RNAi methodologies for livestock species will bring new opportunities to link diversity at the genome level to functional differences in health traits in livestock species.


title: An 81 base-pair deletion in SARS-CoV-2 ORF7a identified from sentinel surveillance in Arizona (Jan-Mar 2020)

text: On January 26 2020, the first Coronavirus Disease 2019 (COVID-19) case was reported in Arizona of an individual with travel history (3rd case in the US). Here, we report on early SARS-CoV-2 sentinel surveillance in Tempe, Arizona. Genomic characterization identified an isolate encoding a 27 amino acid in-frame deletion in accessory protein ORF7a, the ortholog of SARS-CoV immune antagonist ORF7a/X4 .


Editorial: Concepts and Experiences in Framing, Integration and Evaluation of One Health and EcoHealth


title: Evaluation of prognosis related to compliance with supportive periodontal treatment in patients with chronic periodontitis: a clinical retrospective study

text: PURPOSE: The purpose of this study was to evaluate the prognostic effect of patient compliance with supportive periodontal treatment (PC-SPT). Chronic periodontitis patients were classified based on their compliance level, and factors affecting PC-SPT and the prognosis of PC-SPT were investigated. METHODS: This study selected 206 patients who started SPT after receiving periodontal treatment between 2010 and 2012. Patients who continued SPT through February 2016 were included. The patients were classified according to whether they exhibited complete compliance (100% of visits), excellent compliance (≥70% of visits), incomplete compliance (<70% of visits), or non-compliance (only 2 visits). Patient characteristics that could affect PC-SPT, such as age, sex, distance of the clinic from their residence, implantation, and periodontal treatment, were investigated. The number of newly decayed and extracted teeth, alveolar bone level changes around the teeth and implants, and implant removal were examined to evaluate the prognosis of PC-SPT. RESULTS: Sex and the presence of an implant significantly affected PC-SPT. Additionally, the number of newly decayed and extracted teeth and changes in alveolar bone levels around the teeth and implants were significant prognostic factors related to PC-SPT. CONCLUSIONS: PC-SPT in chronic periodontitis patients will help maintain periodontal health and prevent further periodontal disease.


title: Saliva: a diagnostic option and a transmission route for 2019-nCoV

text: Data sources This review included 13 clinical studies (observational or clinical trial) which reported results of studies of the 2019 novel coronavirus (2019-nCoV). The other 62 referenced papers were of different types (eg, reviews, WHO protocols, letter to editor etc). Study selection The study selected trials, reviews, and in-vitro research assessing the critical aspects of saliva as an easily accessible and early-stage diagnostic source, and also an entry route for 2019-nCoV. Most of the clinical studies were descriptive case series of patients who had contracted 2019-nCoV in China. These were mainly studies designed to compare saliva samples with throat swabs, with regard to the presence of 2019-nCoV RNA. Another aspect of the included studies was the susceptibility of oral tissues to 2019-nCoV due to the expression of angiotensin-converting enzyme II (ACE2) as a receptor for the 2019-nCoV. Some review studies and clinical infection control protocols were also included to discuss the transmission patterns of 2019-nCoV from the oral cavity. Studies were not restricted to English language and they were not all full-text papers. Data extraction and synthesis A narrative synthesis of the results was conducted using distinct headings and subheadings, defined by the authors based on relevancy to the consensus about the importance of saliva with reference to 2019-nCoV. Results There was an inherent heterogeneity among the included clinical studies concerning their designs, sampling techniques, and the results about the diagnostic value of saliva. The percentage of coronavirus disease of 2019 (COVID-19) patients with positive 2019-nCoV RNA varied from 12.9% to 91.67% among these studies. Regarding the possibility of direct virus invasion into the oral tissues, the results suggested that ACE2+ cells in salivary glands could possibly be the target cells of 2019-nCoV and theoretically could generate infectious saliva in a sustained way. Furin was suggested as another protein which makes the tongue more vulnerable to 2019-nCoV, especially in conditions inducing its upregulation (for example, squamous cell carcinoma). According to the comparisons between 2019-nCoV and SARS-CoV, saliva could be considered of diagnostic value via the early detection of viral RNA for both of the viruses. Whilst the viral peak was shown to be at onset of symptoms for 2019-nCoV, it can linger up to the tenth day after the appearance of symptoms for SARS-CoV. Finally, this paper warns about airborne transmission, particularly for close contacts. Conclusions Saliva can be proposed as an easily accessible diagnostic source although further clinical studies are required. Given the presence of viral RNA in saliva in the early stages of COVID-19, the recommendations to wear masks to prevent the rapid transmission of infectious droplets into the air, and keep a safe distance from other people are clearly based in evidence.


title: Illumina MiSeq 16S amplicon sequence analysis of bovine respiratory disease associated bacteria in lung and mediastinal lymph node tissue

text: BACKGROUND: Bovine respiratory disease (BRD) is caused by growth of single or multiple species of pathogenic bacteria in lung tissue following stress and/or viral infection. Next generation sequencing of 16S ribosomal RNA gene PCR amplicons (NGS 16S amplicon analysis) is a powerful culture-independent open reference method that has recently been used to increase understanding of BRD-associated bacteria in the upper respiratory tract of BRD cattle. However, it has not yet been used to examine the microbiome of the bovine lower respiratory tract. The objective of this study was to use NGS 16S amplicon analysis to identify bacteria in post-mortem lung and lymph node tissue samples harvested from fatal BRD cases and clinically healthy animals. Cranial lobe and corresponding mediastinal lymph node post-mortem tissue samples were collected from calves diagnosed as BRD cases by veterinary laboratory pathologists and from clinically healthy calves. NGS 16S amplicon libraries, targeting the V3-V4 region of the bacterial 16S rRNA gene were prepared and sequenced on an Illumina MiSeq. Quantitative insights into microbial ecology (QIIME) was used to determine operational taxonomic units (OTUs) which corresponded to the 16S rRNA gene sequences. RESULTS: Leptotrichiaceae, Mycoplasma, Pasteurellaceae, and Fusobacterium were the most abundant OTUs identified in the lungs and lymph nodes of the calves which died from BRD. Leptotrichiaceae, Fusobacterium, Mycoplasma, Trueperella and Bacteroides had greater relative abundances in post-mortem lung samples collected from fatal cases of BRD in dairy calves, compared with clinically healthy calves without lung lesions. Leptotrichiaceae, Mycoplasma and Pasteurellaceae showed higher relative abundances in post-mortem lymph node samples collected from fatal cases of BRD in dairy calves, compared with clinically healthy calves without lung lesions. Two Leptotrichiaceae sequence contigs were subsequently assembled from bacterial DNA-enriched shotgun sequences. CONCLUSIONS: The microbiomes of the cranial lung lobe and mediastinal lymph node from calves which died from BRD and from clinically healthy H-F calves have been characterised. Contigs corresponding to the abundant Leptotrichiaceae OTU were sequenced and found not to be identical to any known bacterial genus. This suggests that we have identified a novel bacterial species associated with BRD. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12917-017-1035-2) contains supplementary material, which is available to authorized users.


title: Chapter fifteen Enzymatic Assembly of Overlapping DNA Fragments

text: Abstract Three methods for assembling multiple, overlapping DNA molecules are described. Each method shares the same basic approach: (i) an exonuclease removes nucleotides from the ends of double-stranded (ds) DNA molecules, exposing complementary single-stranded (ss) DNA overhangs that are specifically annealed; (ii) the ssDNA gaps of the joined molecules are filled in by DNA polymerase, and the nicks are covalently sealed by DNA ligase. The first method employs the 3′-exonuclease activity of T4 DNA polymerase (T4 pol), Taq DNA polymerase (Taq pol), and Taq DNA ligase (Taq lig) in a two-step thermocycled reaction. The second method uses 3′-exonuclease III (ExoIII), antibody-bound Taq pol, and Taq lig in a one-step thermocycled reaction. The third method employs 5′-T5 exonuclease, Phusion® DNA polymerase, and Taq lig in a one-step isothermal reaction and can be used to assemble both ssDNA and dsDNA. These assembly methods can be used to seamlessly construct synthetic and natural genes, genetic pathways, and entire genomes and could be very useful for molecular engineering tools.


title: Pandemics: good hygiene is not enough

text: The US government is doing well to communicate uncertainty over swine flu. It must also help the public to visualize what a bad pandemic might be like, says Peter M. Sandman.


title: Psychological status and behavior changes of the public during the COVID-19 epidemic in China

text: BACKGROUND: A cluster of pneumonia cases were reported by Wuhan Municipal Health Commission, China in December 2019. A novel coronavirus was eventually identified, and became the COVID-19 epidemic that affected public health and life. We investigated the psychological status and behavior changes of the general public in China from January 30 to February 3, 2020. METHODS: Respondents were recruited via social media (WeChat) and completed an online questionnaire. We used the State-Trait Anxiety Inventory, Self-rating Depression Scale, and Symptom Checklist-90 to evaluate psychological status. We also investigated respondents’ behavior changes. Quantitative data were analyzed by t-tests or analysis of variance, and classified data were analyzed with chi-square tests. RESULTS: In total, 608 valid questionnaires were obtained. More respondents had state anxiety than trait anxiety (15.8% vs 4.0%). Depression was found among 27.1% of respondents and 7.7% had psychological abnormalities. About 10.1% of respondents suffered from phobia. Our analysis of the relationship between subgroup characteristics and psychological status showed that age, gender, knowledge about COVID-19, degree of worry about epidemiological infection, and confidence about overcoming the outbreak significantly influenced psychological status. Around 93.3% of respondents avoided going to public places and almost all respondents reduced Spring Festival-related activities. At least 70.9% of respondents chose to take three or more preventive measures to avoid infection. The three most commonly used prevention measures were making fewer trips outside and avoiding contact (98.0%), wearing a mask (83.7%), and hand hygiene (82.4%). CONCLUSIONS: We need to pay more attention to public psychological stress, especially among young people, as they are likely to experience anxiety, depression, and psychological abnormalities. Different psychological interventions could be formulated according to the psychological characteristics of different gender and age groups. The majority of respondents followed specific behaviors required by the authorities, but it will take time to observe the effects of these behaviors on the epidemic.


title: Hepatitis C Virus Resistance to Carbohydrate-Binding Agents

text: Carbohydrate binding agents (CBAs), including natural lectins, are more and more considered as broad-spectrum antivirals. These molecules are able to directly inhibit many viruses such as Human Immunodeficiency Virus (HIV), Hepatitis C Virus (HCV), Dengue Virus, Ebola Virus or Severe Acute Respiratory Syndrome Coronavirus through binding to envelope protein N-glycans. In the case of HIV, it has been shown that CBAs select for mutant viruses with N-glycosylation site deletions which are more sensitive to neutralizing antibodies. In this study we aimed at evaluating the HCV resistance to CBAs in vitro. HCV was cultivated in the presence of increasing Galanthus nivalis agglutinin (GNA), Cyanovirin-N, Concanavalin-A or Griffithsin concentrations, during more than eight weeks. At the end of lectin exposure, the genome of the isolated strains was sequenced and several potential resistance mutations in the E1E2 envelope glycoproteins were identified. The effect of these mutations on viral fitness as well as on sensitivity to inhibition by lectins, soluble CD81 or the 3/11 neutralizing antibody was assessed. Surprisingly, none of these mutations, alone or in combination, conferred resistance to CBAs. In contrast, we observed that some mutants were more sensitive to 3/11 or CD81-LEL inhibition. Additionally, several mutations were identified in the Core and the non-structural proteins. Thus, our results suggest that in contrast to HIV, HCV resistance to CBAs is not directly conferred by mutations in the envelope protein genes but could occur through an indirect mechanism involving mutations in other viral proteins. Further investigations are needed to completely elucidate the underlying mechanisms.


Overuse/Abuse of the definition of ‘Aerosol Generating Procedures’ to limit mask use.


title: Comparative Proteome Analysis of Porcine Jejunum Tissues in Response to a Virulent Strain of Porcine Epidemic Diarrhea Virus and Its Attenuated Strain

text: Porcine epidemic diarrhea virus (PEDV), a predominant cause of acute enteric infection, leads to severe dehydrating diarrhea and mortality in piglets all over the world. A virulent PEDV YN13 strain, isolated in our laboratory, was attenuated to yield an attenuated PEDV strain YN144. To better understand the pathogenesis mechanism and the virus-host interaction during infection with both PEDV YN13 and YN144 strains, a comparative proteomic analysis was carried out to investigate the proteomic changes produced in the primary target organ, using isobaric tags for relative and absolute quantitation (iTRAQ) labeling, followed by liquid chromatography tandem-mass spectrometry (LC-MS/MS). A total of 269 and 301 differently expressed proteins (DEPs) were identified in the jejunum tissues of the piglets inoculated with YN13 and YN144, respectively. Bioinformatics analysis revealed that these proteins were involved in stress responses, signal transduction, and the immune system. All of these involved interferon-stimulated genes (ISGs) which were up-regulated in jejunums by both of the PEDV-infected groups. Based on the comparative analysis, we proposed that different changes induced by YN13 and YN144 in heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1), eukaryotic initiation factor 4G1 (eIF4G1), and some members in the heat shock protein (HSP) family, may be responsible for differences in their pathogenicity.


Covid-19: Herd immunity is "unethical and unachievable," say experts after report of 5% seroprevalence in Spain


Adjustments in analyses of vitamin D status, allowing for vitamin D determinants, for Covid‐19 risks


title: Arbitration Privacy and Confidentiality In the Age of (Coronavirus) Technology

text: Privacy is a hallmark attribute of arbitration.


title: Co-movements of returns in the health care sectors from the US, UK, and Germany stock markets: Evidence from the continuous wavelet analyses

text: Abstract As the health care sector is one of the largest and fastest growing industries around the world, this study utilizes the wavelet approach to investigate the co-movement of returns in this sector from the US, UK, and Germany stock markets over the period of 1992–2013. Our results suggest that the return of the health care sector in the UK (US) stock market leads those in the US and Germany (Germany) stock markets in the short run and medium run, while the returns of the health care sector in the US stock market lead those in the UK stock market in the long run. We find the least (most) structural change in the return of the health care sector for the UK (US) stock market, as both countries belong to two extreme (i.e., public and private) health care systems. There is also an overall increase of long-run interdependence between UK and US health care sectors. Both the subprime mortgage crisis and global financial crisis caused clear contagion effects between the health care sectors of DE and UK as well as those of US and DE. Finally, our results highlight the importance of taking into consideration the time and frequency-varying properties of health care sectors’ stock return co-movement in international portfolios.


title: Recent Advances in Aptamer Discovery and Applications

text: Aptamers are short, single-stranded DNA, RNA, or synthetic XNA molecules that can be developed with high affinity and specificity to interact with any desired targets. They have been widely used in facilitating discoveries in basic research, ensuring food safety and monitoring the environment. Furthermore, aptamers play promising roles as clinical diagnostics and therapeutic agents. This review provides update on the recent advances in this rapidly progressing field of research with particular emphasis on generation of aptamers and their applications in biosensing, biotechnology and medicine. The limitations and future directions of aptamers in target specific delivery and real-time detection are also discussed.


title: CSLR - EU Update - April 2020

text: Abstract Relevant DLA Piper articles published since the previous submission in February


title: Propidium monoazide (PMA) and ethidium bromide monoazide (EMA) improve DNA array and high-throughput sequencing of porcine reproductive and respiratory syndrome virus identification

text: Abstract Pan-viral DNA array (PVDA) and high-throughput sequencing (HTS) are useful tools to identify novel viruses of emerging diseases. However, both techniques have difficulties to identify viruses in clinical samples because of the host genomic nucleic acid content (hg/cont). Both propidium monoazide (PMA) and ethidium bromide monoazide (EMA) have the capacity to bind free DNA/RNA, but are cell membrane-impermeable. Thus, both are unable to bind protected nucleic acid such as viral genomes within intact virions. However, EMA/PMA modified genetic material cannot be amplified by enzymes. In order to assess the potential of EMA/PMA to lower the presence of amplifiable hg/cont in samples and improve virus detection, serum and lung tissue homogenates were spiked with porcine reproductive and respiratory virus (PRRSV) and were processed with EMA/PMA. In addition, PRRSV RT-qPCR positive clinical samples were also tested. EMA/PMA treatments significantly decreased amplifiable hg/cont and significantly increased the number of PVDA positive probes and their signal intensity compared to untreated spiked lung samples. EMA/PMA treatments also increased the sensitivity of HTS by increasing the number of specific PRRSV reads and the PRRSV percentage of coverage. Interestingly, EMA/PMA treatments significantly increased the sensitivity of PVDA and HTS in two out of three clinical tissue samples. Thus, EMA/PMA treatments offer a new approach to lower the amplifiable hg/cont in clinical samples and increase the success of PVDA and HTS to identify viruses.


title: Systemic treatment of patients with gastrointestinal cancers during the COVID-19 outbreak: COVID-19-adapted recommendations of the National Cancer Institute of Milan

text: The current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak poses a major challenge in the treatment decision making of cancer patients, who may be at higher risk of developing a severe and deadly SARS-CoV-2 infection compared to the general population. The healthcare emergency is forcing to reshape the daily assessment between risks and benefits expected from the administration of immune-suppressive and potentially toxic treatments. To guide our clinical decisions at the National Cancer Institute of Milan (Lombardy region, the epicentre of the outbreak in Italy), we formulated Coronavirus-adapted Institutional recommendations for the systemic treatment of patients with gastrointestinal cancers. Here we describe how our daily clinical practice has changed due to the pandemic outbreak, with the aim of providing useful suggestions for physicians that are facing the same challenges worldwide.


title: Independent research of China in Science Citation Index Expanded during 1980–2011

text: Abstract The study explores the characteristics of China's independent research articles published from 1980 to 2011, based on the database of Science Citation Index Expanded. The publication outputs of seven major industrialized countries including Canada, France, Japan, Germany, Italy, the UK, and the USA were compared with China. Annual production, field performance, research emphases and trends, top articles, as well as main institutional and individual contributors by its top cited articles were analyzed. Some newly developed indicators related to words in title, author keywords, KeyWords Plus, first author, corresponding author, and Y-index were employed to provide in-depth information on topic and author contributions. Results showed that China has been closing the gap with the USA with the greatest growth, and has stood the second since 2006. Most top cited articles were published in 2000s, made up approximately seven tenths of total articles. Pronounced activities were found in chemistry and physics related categories. The core categories included multidisciplinary chemistry, physical chemistry, multidisciplinary materials science, and applied physics. Moreover, China's performance of nanotechnology and science, especially carbon nanotubes, nanoparticles, nanowires, and nanostructures showed dramatic growth. Six top articles with at least 1000 citations were examined, and were observed to concern medicine, nanotube, and adsorption. In addition, main contributing institutions and authors were also revealed and evaluated. Chinese Academy of Sciences played a dominant role, and Tsinghua University, Peking University and five universities in Hong Kong showed good scientific performance.


title: Probability of introducing porcine epidemic diarrhea virus into Danish pig herds by imported spray-dried porcine plasma

text: BACKGROUND: Porcine epidemic diarrhea virus (PEDV) has never been reported in Denmark, but it has been found in Europe, Asia and North America. Ultimately, PEDV has been associated with devastating outbreaks in pig farms. We developed a stochastic simulation model to carry out a quantitative risk assessment and to estimate the annual probability (PPlasma) of introducing PEDV into the Danish pig population, by imported spray-dried porcine plasma (SDPP). The model was based on information from literature and Danish feed companies. Moreover testing the batch of raw blood (before the spray-drying) was considered as potential risk mitigation measure in the future. RESULTS: The median PPlasma was 0.2 % (90 % P.I.: 0.003 %; 2.6 %). Hence, the annual probability of introducing PEDV into the Danish pig population by imported SDPP appeared very low, and on average at least one introduction each 500 years – corresponding to 1/0.002 - could be expected. However, if PEDV survived the spray-drying process and storage was insufficient to completely remove the remaining viable virus (e.g. due to storage at low environmental temperatures during a short time period) the PPlasma was 4.7 % (0.06 %; 57.4 %). In that case, on average, at least one PEDV introduction each 21 years could be expected. This probability could be reduced to 0.3 % (0.004 %; 6.0 %) if the raw batch of blood could be tested before drying (corresponding to at least one introduction each 333 years on average). CONCLUSIONS: This study provides preliminary and important information on the probability of introducing PEDV into the Danish pig population by use of SDPP. Currently PED is not a notifiable disease in the EU and uncertainty was present in our estimates due to possible underreporting in EU Member States, from which SDPP is imported into Denmark. In the future, PED might become a notifiable disease, and in such a case, new knowledge could become available on its epidemiology. Moreover, SDPP could be imported more safely if: producers find a way to substantiate freedom from disease (at least) in herds delivering blood for SDPP, the batch of blood tests negative for PEDV and conditions for processing/storage required by the international laws are respected.


title: Coumarin as a structural component of substrates and probes for serine and cysteine proteases

text: Coumarins represent well-established structures to introduce fluorescence into tool compounds for biochemical investigations. They are valued for their small size, chemical stability and accessibility as well as their tunable photochemical properties. As components of fluorophore/quencher pairs or FRET donor/acceptor pairs, coumarins have frequently been applied in substrate mapping approaches for serine and cysteine proteases. This review also focuses on the incorporation of coumarins into the side chain of amino acids and the exploitation of the resulting fluorescent amino acids for the positional profiling of protease substrates. The protease-inhibiting properties of certain coumarin derivatives and the utilization of coumarin moieties to assemble activity-based probes for serine and cysteine protease are discussed as well.


title: Immune Protection of Nonhuman Primates against Ebola Virus with Single Low-Dose Adenovirus Vectors Encoding Modified GPs

text: BACKGROUND: Ebola virus causes a hemorrhagic fever syndrome that is associated with high mortality in humans. In the absence of effective therapies for Ebola virus infection, the development of a vaccine becomes an important strategy to contain outbreaks. Immunization with DNA and/or replication-defective adenoviral vectors (rAd) encoding the Ebola glycoprotein (GP) and nucleoprotein (NP) has been previously shown to confer specific protective immunity in nonhuman primates. GP can exert cytopathic effects on transfected cells in vitro, and multiple GP forms have been identified in nature, raising the question of which would be optimal for a human vaccine. METHODS AND FINDINGS: To address this question, we have explored the efficacy of mutant GPs from multiple Ebola virus strains with reduced in vitro cytopathicity and analyzed their protective effects in the primate challenge model, with or without NP. Deletion of the GP transmembrane domain eliminated in vitro cytopathicity but reduced its protective efficacy by at least one order of magnitude. In contrast, a point mutation was identified that abolished this cytopathicity but retained immunogenicity and conferred immune protection in the absence of NP. The minimal effective rAd dose was established at 10(10) particles, two logs lower than that used previously. CONCLUSIONS: Expression of specific GPs alone vectored by rAd are sufficient to confer protection against lethal challenge in a relevant nonhuman primate model. Elimination of NP from the vaccine and dose reductions to 10(10) rAd particles do not diminish protection and simplify the vaccine, providing the basis for selection of a human vaccine candidate.


title: Development of a recombinase polymerase amplification assay with lateral flow dipstick for rapid detection of feline parvovirus

text: Abstract Feline panleukopenia caused by feline parvovirus (FPV), a single-stranded DNA virus, is typically highly contagious and often presents with lethal syndrome. The broad spectrum of possible hosts suggests its potential for transmission from animal to person through close contact with pets. FPV thus serves as an example of the importance of new rapid point-of-care field diagnostic tools for the control and prevention of transmission, especially among rare wild animals and pet cats. Recombinase polymerase amplification (RPA), as a real-time and isothermal method, could be a more affordable alternative to PCR when combined with a lateral flow dipstick (LFD) indicator. In this study, we report a novel FPV lateral flow dipstick RPA (LFD-RPA) instant detection method capable of detecting a range of different FPV strains. The LFD-RPA assay consists of specific primers, probe, and nucleic acid strip. It is capable of detecting 102 copies of target nucleic acid per reaction, which is one order of magnitude higher than the sensitivity of traditional PCR. The most suitable reaction conditions for this assay are at 38 ℃ for 15 min. This paper develops an efficient visual detection system that can eliminate the need for professional staff and expensive and sophisticated equipment for field detection.


title: Chapter 7 Pathogenesis

text: Pathogenicity is the capacity of an organism to cause disease. During virus infections, diseases symptoms arise from two causes, direct injury caused by virus replication and the side effects of the immune response to infection. The balance between these two is variable. The majority of virus infections are asymptomatic and do not cause disease. Only a tiny number of infections cause serious or life-threatening consequences such as immunodeficiency, or tumors. Diseases caused by previously unknown viruses continue to be regularly identified. Occasionally, known viruses appear to change their behavior, suddenly causing outbreaks of diseases. Such viruses are known as emerging viruses and frequently the cause of a new disease is when a virus switches host species and begins to infect another, and changes in human activities also result in the emergence of new or previously unrecognized diseases.


title: Do superspreaders generate new superspreaders? a hypothesis to explain the propagation pattern of COVID-19

text: Abstract The current global propagation of COVID-19 is heterogeneous, with slow transmission continuing in many countries, and exponential propagation in others, in which the time that took to begin this explosive spread varies greatly. It is proposed that this could be explained by cascading superspreading events, in which new infections caused by a superspreader are more likely to be highly infectious. The mechanism suggested for this is related to viral loads. Exposure to high viral loads may result in infections of high intensity, which exposes new cases to high viral loads, and so on. This notion is supported by experimental veterinary research.


title: Impact of city lockdown on the air quality of COVID-19-hit of Wuhan city

text: Abstract A series of strict lockdown measures were implemented in the areas of China worst affected by coronavirus disease 19, including Wuhan, to prevent the disease spreading. The lockdown had a substantial environmental impact, because traffic pollution and industrial emissions are important factors affecting air quality and public health in the region. After the lockdown, the average monthly air quality index (AQI) in Wuhan was 59.7, which is 33.9% lower than that before the lockdown (January 23, 2020) and 47.5% lower than that during the corresponding period (113.6) from 2015 to 2019. Compared with the conditions before the lockdown, fine particulate matter (PM2.5) decreased by 36.9% and remained the main pollutant. Nitrogen dioxide (NO2) showed the largest decrease of approximately 53.3%, and ozone (O3) increased by 116.6%. The proportions of fixed-source emissions and transported external-source emissions in this area increased. After the lockdown, O3 pollution was highly negatively correlated with the NO2 concentration, and the radiation increase caused by the PM2.5 reduction was not the main reason for the increase in O3. This indicates that the generation of secondary pollutants is influenced by multiple factors and is not only governed by emission reduction.


Innovative transthoracic echocardiographic imaging on prone ventilated patients with COVID-19 using a transesophageal probe


title: Cellular pathology in the COVID-19 era: a European perspective on maintaining quality and safety

text: COVID-19 is a zoonotic viral infection that originated in Wuhan, China, in late 2019. WHO classified the resulting pandemic as a ‘global health emergency’ due to its virulence and propensity to cause acute respiratory distress syndrome. The COVID-19 pandemic has had a major impact on diagnostic laboratories, particularly those handling cell and tissue specimens. This development carries serious implications for laboratory practice in that safety of personnel has to be balanced against high-quality analysis and timely reporting of results. The aim of this article is to present some recommendations for the handling of such specimens in the preanalytical, analytical and postanalytical phases of laboratory testing and analysis in an era of high COVID-19 prevalence, such as that seen, for example, in the UK, Spain, Italy and France.


A Post‐COVID‐19 Parkinsonism in the Future?


title: Impact of the COVID-19 pandemic on headache management in Spain: an analysis of the current situation and future perspectives

text: Abstract Introduction The COVID-19 pandemic has had a great impact on healthcare systems. Spain, where headache is the main reason for outpatient neurology consultation, is one of the countries with the most reported cases of the disease. Objectives This study aimed to analyse the impact of the COVID-19 pandemic on headache units in Spain and to evaluate how neurologists see the future of these units. Methods We conducted a cross-sectional online survey of headache units during the sixth week of the state of alarm declared in Spain in response to the pandemic. Results The response rate was 74%, with the participation of centres with different characteristics and from all Autonomous Communities of Spain. Limitations in face-to-face activity were reported by 95.8% of centres, with preferential face-to-face consultation being maintained in 60.4%, and urgent procedures in 45.8%. In 91.7% of centres, the cancelled face-to-face activity was replaced by telephone consultation. 95.8% of respondents stated that they would use personal protection equipment in the future, and 86% intended to increase the use of telemedicine. The majority foresaw an increase in waiting lists (93.8% for initial consultations, 89.6% for follow-up, and 89.4% for procedures) and a worse clinical situation for patients, but only 15% believed that their healthcare structures would be negatively affected in the future. Conclusions As a consequence of the pandemic, headache care and research activity has reduced considerably. This demonstrates the need for an increase in the availability of telemedicine in our centres in the near future.


title: The impact of SARS on epilepsy: The experience of drug withdrawal in epileptic patients

text: Summary Background During the 2003 severe acute respiratory syndrome (SARS) outbreak, many patients avoided hospital visit because of fear of infection. Antiepileptic drug (AED) withdrawal is a risk factor for seizure recurrence. Therefore, seizure control during the SARS outbreak is a good model for examining the impact of drug withdrawal in seizure control. Methods All seizures experienced by each patient before, during, and after the SARS outbreak periods were registered in each patient's seizure diary. The patients were divided into four groups according to the presence of drug withdrawal as well as seizure attack. In each group, seizures occurring during three different periods were compared. Risk factors for seizure recurrence were also examined. Results Of 227 cases, 49 stopped taking medication during the outbreak. Among them, 28 suffered seizure attacks during AED withdrawal. Four cases developed cluster attacks and two cases had status epilepticus after AED withdrawal. AED withdrawal produced a significant increase in seizure frequency. The major risk factors for withdrawal seizures were symptomatic etiologies, polytherapy and non-seizure free before AED withdrawal. Conclusions The SARS outbreak adversely affected seizure control because of AED withdrawal. Patients with polytherapy, non-seizure free and symptomatic etiologies were more susceptible to recurrence of seizures after AED withdrawal.


title: The SARS epidemic in mainland China: bringing together all epidemiological data

text: Objective To document and verify the number of cases of severe acute respiratory syndrome (SARS) during the 2002–2003 epidemic in mainland China. Method All existing Chinese SARS data sources were integrated in one final database. This involved removing non‐probable and duplicate cases, adding cases at the final stage of the outbreak, and collecting missing information. Results The resulting database contains a total of 5327 probable SARS cases, of whom 343 died, giving a case fatality ratio (CFR) of 6.4%. While the total number of cases happens to be equal to the original official reports, there are 5 cases overall which did not result in death. When compared with Hong Kong Special Administrative Region of China, China Taiwan, and Singapore, the SARS epidemic in mainland China resulted in a considerably lower CFR, involved relatively younger cases and included fewer health care workers. Conclusions To optimise future data collection during large‐scale outbreaks of emerging or re‐emerging infectious disease, China must further improve the infectious diseases reporting system, enhance collaboration between all levels of disease control, health departments, hospitals and institutes nationally and globally, and train specialized staff working at county centres of disease control.


title: Isolation and Identification of a Natural Reassortant Mammalian Orthoreovirus from Least Horseshoe Bat in China

text: BACKGROUND: Mammalian orthoreoviruses (MRVs) have a wide geographic distribution and can infect virtually all mammals. Infections in humans may be either symptomatic or asymptomatic. This study describes the isolation and identification of a natural reassortant MRV from least horseshoe bats (Rhinolophus pusillu) in China, referred to as RpMRV-YN2012. METHODS AND RESULTS: The RpMRV-YN2012 was obtained from urine samples of Rhinolophus pusillus by cell culture. Negative-staining electron microscopy revealed that RpMRV-YN2012 was a non-enveloped icosahedral virus with ∼75 nm in diameter. Polyacrylamide gel electrophoresis (PAGE) migration patterns of the genome segments showed that RpMRV-YN2012 contained 10 segments in a 3:3:4 arrangement. The whole genome sequence of RpMRV2012 was determined. The consensus terminal sequences of all segments of 5’-GCUAh…yUCAUC-3’ (h = A, U or C; y = C or U) were similar to the MRV species within the genus Orthoreovirus. Its evolution and evidence of genetic reassortment were analyzed by sequence comparison and phylogenetic analysis. The results showed that RpMRV-YN2012 is a novel serotype 2 MRV that may have originated from reassortment among bat, human, and/or pig MRV strains which associated with diarrhea, acute gastroenteritis and necrotizing encephalopathy in animals and humans. CONCLUSIONS: RpMRV-YN2012 is a novel bat reassortant MRV, which may have resulted from a reassortment involving MRVs known to infect humans and animals. It is necessary to identify whether RpMRV-YN2012 is associated with diarrhea, acute gastroenteritis and necrotizing encephalopathy in clinical patients. In addition, we should carefully monitor its evolution and virulence in real time.


title: Climate Change Could Increase the Geographic Extent of Hendra Virus Spillover Risk

text: Disease risk mapping is important for predicting and mitigating impacts of bat-borne viruses, including Hendra virus (Paramyxoviridae:Henipavirus), that can spillover to domestic animals and thence to humans. We produced two models to estimate areas at potential risk of HeV spillover explained by the climatic suitability for its flying fox reservoir hosts, Pteropus alecto and P. conspicillatus. We included additional climatic variables that might affect spillover risk through other biological processes (such as bat or horse behaviour, plant phenology and bat foraging habitat). Models were fit with a Poisson point process model and a log-Gaussian Cox process. In response to climate change, risk expanded southwards due to an expansion of P. alecto suitable habitat, which increased the number of horses at risk by 175–260% (110,000–165,000). In the northern limits of the current distribution, spillover risk was highly uncertain because of model extrapolation to novel climatic conditions. The extent of areas at risk of spillover from P. conspicillatus was predicted shrink. Due to a likely expansion of P. alecto into these areas, it could replace P. conspicillatus as the main HeV reservoir. We recommend: (1) HeV monitoring in bats, (2) enhancing HeV prevention in horses in areas predicted to be at risk, (3) investigate and develop mitigation strategies for areas that could experience reservoir host replacements. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10393-018-1322-9) contains supplementary material, which is available to authorized users.


title: Therapeutic effect of anti-feline TNF-alpha monoclonal antibody for feline infectious peritonitis

text: Abstract Feline infectious peritonitis virus (FIPV) replication in macrophages/monocytes induced tumor necrosis factor (TNF)-alpha production, and that the TNF-alpha produced was involved in aggravating the pathology of FIP. We previously reported the preparation of a feline TNF-alpha (fTNF-alpha)-neutralizing mouse monoclonal antibody (anti-fTNF-alpha mAb). This anti-fTNF-alpha mAb 2–4 was confirmed to inhibit the following fTNF-alpha-induced conditions in vitro. In the present study, we investigated whether mAb 2–4 improved the FIP symptoms and survival rate of experimentally FIPV-inoculated SPF cats. Progression to FIP was prevented in 2 out of 3 cats treated with mAb 2–4, whereas all 3 cats developed FIP in the placebo control group. Plasma alpha1-glycoprotein and vascular endothelial growth factor levels were improved by the administration of mAb 2–4, and the peripheral lymphocyte count also recovered. These results strongly suggested that the anti-fTNF-alpha antibody is effective for the treatment of FIP.


title: Broad respiratory virus detection in infants hospitalized for bronchiolitis by use of a multiplex RT‐PCR DNA microarray system

text: Newly available molecular tools allow a sensitive detection of a broad panel of viruses in respiratory tract specimens. In the present study, the application of a multiplex RT‐PCR DNA microarray in diagnosis and epidemiological survey of viral infections in infants hospitalized for bronchiolitis was assessed. One hundred and thirty‐eight nasopharyngeal aspirates collected from October 2007 to September 2008 were tested by direct immunofluorescence and viral culture, a combination of referenced RT‐PCRs and the DNA microarray. One or more viruses were detected in 96, 126 and 126 of the specimens by direct immunofluorescence and viral culture, RT‐PCRs and DNA microarray, respectively (70 vs. 91 vs. 91%, P < 10(−3)). The RT‐PCRs and the DNA microarray yielded concordant results for 99% of specimens and identified mixed viral infections in 85 (62%). The most common associations were: human bocavirus and respiratory syncytial virus (32%), adenovirus and respiratory syncytial virus (30%), and parainfluenza virus type 3 and respiratory syncytial virus (23%). None of the bronchiolitis severity parameters including intensive care unit admission, O(2) supply, O(2) saturation percentage, O(2) length and length of stay at the hospital appeared to be significantly increased in multiple viral infections compared to single viral infections (P > 0.1). In conclusion, the use of this DNA microarray in clinical virology practice allows rapid and accurate identification of common and uncommon viral respiratory pathogens in infants hospitalized for bronchiolitis. It should improve the clinical management, the epidemiological survey, and the prevention of the nosocomial transmission of respiratory viruses in pediatric wards. J. Med. Virol. 84:979–985, 2012. © 2012 Wiley Periodicals, Inc.


title: Optimizing the design of protein nanoparticles as carriers for vaccine applications

text: Successful vaccine development remains a huge challenge for infectious diseases such as malaria, HIV and influenza. As a novel way to present antigenic epitopes to the immune system, we have developed icosahedral self-assembling protein nanoparticles (SAPNs) to serve as a prototypical vaccine platform for infectious diseases. Here we examine some biophysical factors that affect the self-assembly of these nanoparticles, which have as basic building blocks coiled-coil oligomerization domains joined by a short linker region. Relying on in silico computer modeling predictions, we selected five different linker regions from the RCSB protein database that connect oligomerization domains, and then further studied the self-assembly and stability of in vitro produced nanoparticles through biophysical characterization of formed particles. One design in particular, T2i88, revealed excellent self-assembly and homogeneity thus paving the way towards a more optimized nanoparticle for vaccine applications.


title: Compact optical diagnostic device for isothermal nucleic acids amplification

text: Abstract We recently reported the successful use of the loop-mediated isothermal amplification (LAMP) reaction for hepatitis B virus (HBV) DNA amplification and its optimal primer design method. In this study, we report the development of an integrated isothermal device for both amplification and detection of targeted HBV DNA. It has two major components, a disposable polymethyl methacrylate (PMMA) micro-reactor and a temperature-regulated optical detection unit (base apparatus) for real-time monitoring of the turbidity changes due to the precipitation of DNA amplification by-product, magnesium pyrophosphate. We have established a correlation curve (R 2 =0.99) between the concentration of pyrophosphate ions and the level of turbidity by using a simulated chemical reaction to evaluate the characteristics of our device. For the applications of rapid pathogens detection, we also have established a standard curve (R 2 =0.96) by using LAMP reaction with a standard template in our device. Moreover, we also have successfully used the device on seven clinical serum specimens where HBV DNA levels have been confirmed by real-time PCR. The result indicates that different amounts of HBV DNA can be successfully detected by using this device within 1h.


title: Overview of transnational recommendations for COVID‐19 transmission control in dental care settings

text: On 11 March 2020, the World Health Organization (WHO) declared the coronavirus disease (COVID‐19) caused by severe acute respiratory syndrome coronavirus (SARS‐CoV‐2) as a pandemic. Until an effective treatment or a vaccine is developed, the current recommendations are to contain the disease, and control its transmission. It is now clear that the primary mode of SARS‐CoV‐2 transmission is aerosol/droplet spread, and by contacting virus‐contaminated surfaces acting as fomites (inanimate vectors). Furthermore, recent data indicate that the live virus particles are present in saliva, and, more alarmingly, asymptomatic individuals may transmit the infection. By virtue of the nature of the practice of dentistry where intrinsically, a high volume of aerosols is produced, as well as the close proximity of dentists and patients during treatment, dentists and allied health staff are considered the highest risk health professional group for acquiring SARS‐CoV‐2 during patient management. Therefore, several organizations and specialty associations have proposed guidelines and recommendations for limiting the transmission of SARS‐COV‐2 from carriers to dentists and vice versa. This paper aims to provide a review of these guidelines, and concludes with a brief look at how the practice of dentistry may be impacted by COVID‐19, in the post‐pandemic era.


title: Aminopeptidase-N-independent entry of porcine epidemic diarrhea virus into Vero or porcine small intestine epithelial cells

text: Abstract A monkey cell line Vero (ATCC CCL-81) is commonly used for porcine epidemic diarrhea virus (PEDV) propagation in vitro. However, it is still controversial whether the porcine aminopeptidase N (pAPN) counterpart on Vero cells (Vero-APN) confers PEDV entry. We found that endogenous expression of Vero-APN was undetectable in the mRNA and the protein levels in Vero cells. We cloned the partial Vero-APN gene (3340-bp) containing exons 1 to 9 from cellular DNA and subsequently generated two APN-knockout Vero cell lines by CRISPR/Cas9 approach. PEDV infection of two APN-knockout Vero cells had the same efficiency as the Vero cells with or without neuraminidase treatment. A Vero cells stably expressing pAPN did not increase PEDV production. SiRNA-knockdown of pAPN in porcine jejunum epithelial cells had no effects on PEDV infection. The results suggest that there exists an additional cellular receptor on Vero or porcine jejunal cells independent of APN for PEDV entry.


title: Wearable Technology and How This Can Be Implemented into Clinical Practice

text: PURPOSE OF REVIEW: Our day-to-day life is saturated with health data that was previously out of reach. Over the last decade, new devices and fitness technology companies are attempting to tap into this data, uncovering a treasure trove of useful information that, when applied correctly, has the potential to revolutionize the way we approach healthcare and chronic conditions like asthma, especially in the wake of the COVID-19 pandemic. RECENT FINDINGS: By harnessing exciting developments in personalization, digitization, wellness, and patient engagement, care providers can improve health outcomes for our patients in a way we have never been able to do in the past. While new technologies to capture individual health metrics are everywhere, how can we use this information to make a real difference in our patients’ lives? Navigating the complicated landscape of personal wearable devices, asthma inhaler sensors, and exercise apps can be daunting to even the most tech savvy physician. SUMMARY: This manuscript will give you the tools necessary to make lasting changes in your patients’ lives by exposing them to a world of usable, affordable, and relatable health technology that resonates with their personal fitness and wellness goals. These tools will be even more important post-COVID-19, as the landscape of clinical outpatient care changes from mainly in-person visits to a greater reliance on telemedicine and remote monitoring.


title: Identification of a novel orthohepadnavirus in pomona roundleaf bats in China

text: Bats in Myanmar, Gabon, and Panama have been found to harbor diverse hepadnaviruses. Here, we report a novel hepadnavirus in 4 of 20 pomona roundleaf bats from Yunnan province, China. This virus contains 3,278 nucleotides (nt) in the full circularized genome, with four predicted open frames (ORFs) reading in the same direction. Full genomic sequence comparisons and evolutionary analysis indicate that this virus is a member of a new species within the genus Orthohepadnavirus


title: Symposium review: Intravital imaging of the lactating mammary gland in live mice reveals novel aspects of milk-lipid secretion

text: ABSTRACT Milk fat comprises membrane-coated droplets of neutral lipid, which constitute the predominant source of lipids for survival of the suckling neonate. From the perspective of the dairy industry, they are the basis for the manufacture of butter and essential ingredients in the production of cheese, yogurt, and specialty dairy produce. To provide mechanistic insight into the assembly and secretion of lipid droplets during lactation, we developed novel intravital imaging techniques using transgenic mice, which express fluorescently tagged marker proteins. The number 4 mammary glands were surgically prepared under a deep plane of anesthesia and the exposed glands positioned as a skin flap with intact vascular supply on the stage of a laser-scanning confocal microscope. Lipid droplets were stained by prior exposure of the glands to hydrophobic fluorescent BODIPY (boron-dipyrromethene) dyes and their formation and secretion monitored by time-lapse subcellular microscopy over periods of 1 to 2 h. Droplets were transported to the cell apex by directed (superdiffusive) motion at relatively slow and intermittent rates (0–2 µm/min). Regardless of size, droplets grew by numerous fusion events during transport and as they were budding from the cell enveloped by apical membranes. Surprisingly, droplet secretion was not constitutive but required an injection of oxytocin to induce contraction of the myoepithelium with subsequent release of droplets into luminal spaces. These novel results are discussed in the context of the current paradigm for milk fat synthesis and secretion and as a template for future innovations in the dairy industry.


title: Re-emergence of a genetic outlier strain of equine arteritis virus: Impact on phylogeny

text: Abstract Equine arteritis virus (EAV) is the causative agent of equine viral arteritis (EVA), a respiratory and reproductive disease of equids, which is notifiable in some countries including the Great Britain (GB) and to the OIE. Herein, we present the case of a persistently infected stallion and the phylogenetic tracing of the virus strain isolated. Discussing EAV occurrence and phylogenetic analysis we review features, which may aid to harmonise and enhance the classification of EAV.


title: Heat inactivation decreases the Qualitative Real-time RT-PCR detection rates of clinical samples with high Cycle Threshold Values in COVID-19

text: Abstract SARS-CoV-2 has caused COVID-19 pandemic globally in the beginning of 2020, and Qualitative Real-time RT-PCR has become the gold standard in diagnosis. As SARSCoV-2 with strong transmissibility and pathogenicity, it has become a professional consensus that clinical samples from suspected patients should be heat inactivated at 56°C for 30min before further processing. However, previous studies on the effect of inactivation on Qualitative Real-time RT-PCR were conducted with diluted samples rather than clinical samples. The aim of this study was to investigate whether heatinactivation on clinical samples before detection will affect the accuracy of Qualitative Real-time RT-PCR detection. All 46 throat swab samples from 46 confirmed inpatients were detected by Qualitative Real-time RT-PCR directly, as well as after heat inactivation. Heat-Inactivation has significantly influenced the qualitative detection results on clinical samples, especially weakly positive samples. The results indicate the urgency to establish a more suitable protocol for COVID-19 clinical sample's inactivation.


title: Long-Term Single-Dose Efficacy of a Vesicular Stomatitis Virus-Based Andes Virus Vaccine in Syrian Hamsters

text: Andes virus (ANDV) is highly pathogenic in humans and is the primary etiologic agent of hantavirus cardiopulmonary syndrome (HCPS) in South America. Case-fatality rates are as high as 50% and there are no approved vaccines or specific therapies for infection. Our laboratory has recently developed a replication-competent recombinant vesicular stomatitis virus (VSV)-based vaccine that expressed the glycoproteins of Andes virus in place of the native VSV glycoprotein (G). This vaccine is highly efficacious in the Syrian hamster model of HCPS when given 28 days before challenge with ANDV, or when given around the time of challenge (peri-exposure), and even protects when administered post-exposure. Herein, we sought to test the durability of the immune response to a single dose of this vaccine in Syrian hamsters. This vaccine was efficacious in hamsters challenged intranasally with ANDV 6 months after vaccination (p = 0.025), but animals were not significantly protected following 1 year of vaccination (p = 0.090). The decrease in protection correlated with a reduction of measurable neutralizing antibody responses, and suggests that a more robust vaccination schedule might be required to provide long-term immunity.


title: Data-driven Framework for Delineating Urban Population Dynamic Patterns: Case Study on Xiamen Island, China

text: The effective data mining of social media has become increasingly recognized for its value in informing decision makers of public welfare. However, existing studies do not fully exploit the underlying merit of big data. In this study, we develop a data-driven framework that integrates machine learning with spatial statistics, and then use it on Xiamen Island, China to delineate urban population dynamic patterns based on hourly Baidu heat map data collected from August 25 to September 3, 2017. The results showed that hot grids are primarily clustered along the main street through the downtown area during working days, whereas cold grids are often observed at the edge of the city during the weekend. The mixed use (of commercial and life services, restaurants and snack bars, offices, leisure areas and sports complexes) is the most significant contributing factor. A new cold grid emerged near conference venues before the Brazil, Russia, India, China, and South Africa Summit, revealing the strong effects of regulations on population dynamics and its evolving patterns. This study demonstrates that the proposed data-driven framework might offer new insights into urban population dynamics and its driving mechanism in support of sustainable urban development.


Comprehensive Assessment of Serious Adverse Events Following Immunization by Health Care Providers


title: In vivo and in vitro Models of Demyelinating Diseases

text: The developmental sequence of a neurotropic strain (JHM) of mouse hepatitis virus was examined by transmission electron microscopy and immunocytology. The nucleo-protein core of this coronavirus, which contains RNA of positive polarity and is helical in configuration, becomes incorporated into enveloped particles in the same manner as the nucleocapsids of the orthomyxo- and paramyxoviruses. However, JHM virus is assembled intracellularly by budding at surfaces of smooth membranous vacuoles. A comparison of JHMvirus replication in L2 and 17C1-1 cell lines revealed that L2 cells undergo more rapid cytopathology and cease virus production much sooner than 17C1-1 cells. In L2 cells the accumulation of core material appears to continue after the abrupt cessation of virus assembly. This is evident by the massive cytoplasmic accumulation of structures resembling nucleocapsids, which react with hybridoma antibody to the nucleocapsid antigen as demonstrated by the immunoperoxidase procedure. The current findings are consistent with our previously published demonstration, using cells of neural and other deviation, of the fundamental role of the host cell type in regulating the replication and expression of coronaviruses.


Respiratory tract infections and pneumonia


title: SARS-CoV Pathogenesis Is Regulated by a STAT1 Dependent but a Type I, II and III Interferon Receptor Independent Mechanism

text: Severe acute respiratory syndrome coronavirus (SARS-CoV) infection often caused severe end stage lung disease and organizing phase diffuse alveolar damage, especially in the elderly. The virus-host interactions that governed development of these acute end stage lung diseases and death are unknown. To address this question, we evaluated the role of innate immune signaling in protection from human (Urbani) and a recombinant mouse adapted SARS-CoV, designated rMA15. In contrast to most models of viral pathogenesis, infection of type I, type II or type III interferon knockout mice (129 background) with either Urbani or MA15 viruses resulted in clinical disease outcomes, including transient weight loss, denuding bronchiolitis and alveolar inflammation and recovery, identical to that seen in infection of wildtype mice. This suggests that type I, II and III interferon signaling play minor roles in regulating SARS pathogenesis in mouse models. In contrast, infection of STAT1−/− mice resulted in severe disease, high virus titer, extensive pulmonary lesions and 100% mortality by day 9 and 30 post-infection with rMA15 or Urbani viruses, respectively. Non-lethal in BALB/c mice, Urbani SARS-CoV infection in STAT1−/− mice caused disseminated infection involving the liver, spleen and other tissues after day 9. These findings demonstrated that SARS-CoV pathogenesis is regulated by a STAT1 dependent but type I, II and III interferon receptor independent, mechanism. In contrast to a well documented role in innate immunity, we propose that STAT1 also protects mice via its role as an antagonist of unrestrained cell proliferation.


title: Viral diseases of ferrets

text: Distemper and rabies vaccination are highly recommended because of the almost invariable fatal outcome of these conditions. Vaccination should constitute an important part of a ferret's preventative medicine program. With the current and anticipated development and licensing of new vaccines, practitioners are invited to gain awareness of the latest vaccine information. Establishment of a practice vaccination protocol with regards to the site of administration of rabies and distemper vaccines is paramount to document any future abnormal tissue reactions. Influenza is the most common zoonotic disease that is seen in ferrets. Although it generally is benign in most ferrets, veterinarians must take this condition seriously. The characteristic continuous antigenic variation of this virus may lead to more virulent strains; the recent emergence of avian influenza virus outbreaks; and the increased susceptibility of elderly, young, and immunosuppressed individuals.


title: Covid‐19 and Myeloma: what are the implications for now and in the future?

text: The pandemic has affected every aspect of myeloma care. Immediate focus is minimizing risk of contracting COVID‐19 and the sequelae of infection. However, what does the future hold for our patients? What lessons will be taken forward to tackle myeloma in the fiscally constrained future? If we embrace the challenges that will emerge in the post‐pandemic environment, the treatment delivered to patients could be more cost effective and better tailored than before. Healthcare delivery post‐COVID will not return to how it was, and now is the time to invest in novel strategies to deliver the best possible outcomes for patients.


title: Detection of Human Bocavirus in Children with Acute Respiratory Tract Infections in Lanzhou and Nanjing, China

text: Abstract Objective The aim of this study was to explore the prevalent characteristics of HBoV1 and its co-infection. Methods PCR was used to detect HBoV1-DNA (HBoV1) and other viruses. A multivariate logistic regression model was used to explore possibility of co-detected for related viruses. Results The positivity rates in Nanjing and Lanzhou were 9.38% (74/789) and 11.62% (161/1386), respectively (P<0.05). The HBoV1 positive group was younger than negative group (P<0.05). Seasonal differences were noted, with a higher frequency of infection in December and July. HBoV1-positive children [72.34% (169/235)] were co-infected with other respiratory viruses. Multifactorial analysis showed no correlations between HBoV1 and the clinical classification, region, gender, age, or treatment as an outpatient or in a hospital. Correlations were identified between HBoV1 infections with ADV (OR=1.53, 95% CI 1.03-2.28), RSV (OR=0.71, 95% CI 0.52-0.98), and IFVA (OR=1.77, 95% CI 1.00-3.13). Conclusions Presence of HBoV1 in nasopharyngeal aspirates did not correlate with region or gender, although the prevalence of HBoV1 was higher in younger children. There were no correlations between HBoV1 and other variables, except for the season and ADV, RSV, or IFVA infections.


title: Tilapia Lake Virus Does Not Hemagglutinate Avian and Piscine Erythrocytes and NH(4)Cl Does Not Inhibit Viral Replication In Vitro

text: Tilapia lake virus (TiLV) is a negative-sense single-stranded RNA (-ssRNA) icosahedral virus classified to be the only member in the family Amnoonviridae. Although TiLV segment-1 shares homology with the influenza C virus PB1 and has four conserved motifs similar to influenza A, B, and C polymerases, it is unknown whether there are other properties shared between TiLV and orthomyxovirus. In the present study, we wanted to determine whether TiLV agglutinated avian and piscine erythrocytes, and whether its replication was inhibited by lysosomotropic agents, such as ammonium chloride (NH(4)Cl), as seen for orthomyxoviruses. Our findings showed that influenza virus strain A/Puerto Rico/8 (PR8) was able to hemagglutinate turkey (Meleagris gallopavo), Atlantic salmon (Salmo salar L), and Nile tilapia (Oreochromis niloticus) red blood cells (RBCs), while infectious salmon anemia virus (ISAV) only agglutinated Atlantic salmon, but not turkey or tilapia, RBCs. In contrast to PR8 and ISAV, TiLV did not agglutinate turkey, Atlantic salmon, or tilapia RBCs. qRT-PCR analysis showed that 30 mM NH(4)Cl, a basic lysosomotropic agent, neither inhibited nor enhanced TiLV replication in E-11 cells. There was no difference in viral quantities in the infected cells with or without NH(4)Cl treatment during virus adsorption or at 1, 2, and 3 h post-infection. Given that hemagglutinin proteins that bind RBCs also serve as ligands that bind host cells during virus entry leading to endocytosis in orthomyxoviruses, the data presented here suggest that TiLV may use mechanisms that are different from orthomyxoviruses for entry and replication in host cells. Therefore, future studies should seek to elucidate the mechanisms used by TiLV for entry into host cells and to determine its mode of replication in infected cells.


title: Infectious disease-related laws: prevention and control measures

text: OBJECTIVES: This study examines recently revised Korean government legislation addressing global infectious disease control for public health emergency situations, with the aim of proposing more rational, effective and realistic interpretations and applications for improvement of law. METHODS: The Korea reported its first laboratory-confirmed case of Middle East Respiratory Syndrome (MERS) coronavirus on May 20, 2015. Since the first indexed case, Korean public health authorities enforced many public health measures that were not authorized in the law; the scope of the current law was too limited to cover MERS. Korea has three levels of government: the central government, special self-governing provinces, and si/gun/gu. Unfortunately, the Infectious Disease Control and Prevention Act does not designate the specific roles of each level of government, and does not state how these governmental branches should be vertically integrated in a state of emergency. RESULTS: When thinking about these policy questions, we should be especially concerned about introducing a new act that deals with all matters relevant to emerging infectious diseases. The aim would be to develop a structure that specifies the roles of each level of government, and facilitates the close collaboration among them, then enacting this in law for the prevention and response of infectious disease. CONCLUSIONS: To address this problem, after analyzing the national healthcare infrastructure along with the characteristics of emerging infectious diseases, we propose the revision of the relevant law(s) in terms of governance aspects, emergency medical countermeasure aspects, and the human rights aspect.


Care of haematology patients in a COVID‐19 epidemic


title: Infecciones por el virus de Epstein-Barr y citomegalovirus

text: Resumen La infección por el virus de Epstein-Barr es la causa más frecuente de la mononucleosis infecciosa. Describimos su diagnóstico y los distintos patrones serológicos que se pueden presentar, así como sus principales complicaciones, relación con el desarrollo de neoplasias, síndromes linfoproliferativos y enfermedades autoinmunes. No existe un tratamiento efectivo. El citomegalovirus es la segunda causa de síndrome mononucleósido, la primera de infección congénita en nuestro medio y presenta gran morbimortalidad en los pacientes inmunodeprimidos, fundamentalmente trasplantados de órganos sólidos, médula ósea e infección por el virus de la inmunodeficiencia humana. Su tratamiento se basa en los antivíricos, sobre todo ganciclovir y valganciclovir. Describimos su manejo, efectos adversos, principales alternativas, nuevos fármacos y profilaxis. Abstract Infections by Epstein-Barr virus is the most common cause of infectious mononucleosis. We describe the diagnosis and different serological patterns, as well as its major complications related to the development of malignancies, lymphoproliferative disorders and autoimmune diseases. There is no effective treatment. Cytomegalovirus is the second cause of mononucleosis syndrome and the first of congenital infection in our environment. It has an high morbidity and mortality in immunocompromised patients, mainly solid organ transplants, bone marrow and coinfection by human immunodeficiency virus. Its treatment is based on antivirals, mainly ganciclovir and valganciclovir. We describe its management, adverse effects, new drugs and prophylaxis.


title: Picobirnavirus detection in bovine and buffalo calves from foothills of Himalaya and Central India

text: The present study describes detection of picobirnavirus (PBV) in faecal samples from bovine and buffalo calves employing the polyacrylamide gel electrophoresis (PAGE). A total of 136 faecal samples from buffalo (n = 122) and cow calves (n = 14) exhibiting clinical signs of diarrhoea and from healthy calves were collected during 2007–2010 from subtropical (central India) and tarai area of western temperate Himalayan foothills (Uttarakhand). The dsRNA nature of the virus was confirmed by nuclease treatment (RNase A, RNaseT1 and DNase 1). PAGE results confirmed 3.67% (5/136) positivity for PBV, showing a typical genomic migration pattern with two discrete bands with size of approximately 2.4 and 1.7 kbps for the larger and smaller segments, respectively. Among the five PBV samples identified, three were from buffalo calves and one from cow calf exhibiting clinical signs of acute diarrhoea, while one sample from non-diarrhoeic buffalo calf also showed the presence of PBV. None of the samples showed dual infection of rotavirus and PBV. The preliminary findings indicate sporadic incidences of PBV in bovine calves and emphasize the need for the development of better diagnostics for early detection and genetic characterization of these emerging isolates of farm animals of economic significance.


title: A Recovered Case of COVID-19 Myocarditis and ARDS Treated with Corticosteroids, Tocilizumab, and Experimental AT-001

text: Abstract: We describe the case of a critically-ill patient with myocarditis and severe ARDS related to COVID-19 infection. This case highlights management strategies including the use of corticosteroids, an IL-6 inhibitor, and an aldose reductase inhibitor resulting in complete clinical recovery.


title: Integration of halofuginone lactate treatment and disinfection with p-chloro-m-cresol to control natural cryptosporidiosis in calves

text: Abstract Two field studies were accomplished on a dairy farm in Saxony to compare different strategies for the control of natural cryptosporidiosis in newborn calves. In the first study, 96 newborn calves were allocated to four different groups immediately after birth. Calves of group H and of group HN were treated orally with 120μg/kg body weight (BW) of halofuginone lactate daily during the first seven days of life. Calves of group C and of group CN were treated with a same volume of tap water. As an additional measure, the pens of groups HN and CN were disinfected with 3% Neopredisan 135-1®(p-chloro-m-cresol), the pens of groups C and H remained non-disinfected. Faeces were examined semi-quantitatively for oocyst excretion using carbolfuchsin-staining and the clinical course was recorded. While disinfection alone (group CN) had no effect on oocyst shedding and diarrhoea, treatment with halofuginone lactate (groups H and HN) reduced oocyst shedding and diarrhoea significantly. Combination of treatment and disinfection (group HN) controlled cryptosporidiosis completely during the first two weeks after birth. However, prevalence of diarrhoea and oocyst shedding was higher in the third week of life in group HN than in any of the other groups. This delayed occurrence of cryptosporidiosis was not seen in study 2 when all calves were similarly protected by treatment and specific disinfection. Oocyst shedding was not observed in peripartal cows either by carbolfuchsin staining, ELISA or PCR.


title: Which type of personal protective equipment (PPE) and which method of donning or doffing PPE carries the least risk of infection for healthcare workers?

text: Data sources CENTRAL, MEDLINE, Embase and CINAHL. Study selection Controlled studies (randomised or non-randomised) that evaluated the effect of full-body PPE on healthcare workers (HCW) exposed to highly infectious diseases, assessed which method of donning and doffing PPE was associated with reduced risk of contamination or infection for HCW, and which training methods increased compliance with PPE protocols. Data extraction and synthesis Two reviewers independently screened the titles and abstracts for inclusion of studies. Full text articles were subsequently assessed for eligibility and disagreements were resolved through consensus. Using criteria outlined in the Cochrane Handbook for Systematic Reviews of Interventions, pairs of review authors independently assessed risk of bias for each randomised study and rated each potential source of bias as high, low, or unclear. ROBINS-I tool was used for the assessment of risk of bias in non-randomised intervention studies. Where appropriate, random effects meta-analyses were conducted. Results A total of 24 studies (randomised controlled trials [RCT] [n = 14]; Quasi-RCT [n = 1] and non-randomised design [n=9]) with 2278 participants were included. Included studies compared types of PPE (n = 8), evaluated modified PPE (n = 6), procedures for donning and doffing PPE (n = 8), and types of training (n = 3). Twenty-two studies were simulation studies, of which 18 simulated exposure of HCW to contaminated body fluids using fluorescent markers or harmless microbes and measured contamination outcomes, and four studies provided modified PPE or procedures and measured compliance with donning and doffing procedures. Types of PPE Powered, air-purifying respirator with coverall may protect against the risk of contamination better than a N95 mask and gown (risk ratio [RR] 0.27, 95% confidence interval [CI] 0.17 to 0.43) but was more difficult to don (non-compliance: RR 7.5, 95% CI 1.81 to 31.1). Gowns compared to aprons may protect better against contamination (MD) -10.28, 95% CI -14.77 to -5.79). Breathable types of PPE are more comfortable and may increase user satisfaction, however with little impact on contamination. Modified PPE versus standard PPE Appropriate modifications to PPE design may lead to less contamination compared to standard PPE. For example, contamination can be reduced using a sealed gown and glove combination so that they can be removed together and cover the wrist area (RR 0.27, 95% CI 0.09 to 0.78), tight fitting gown around the neck, wrist area and hands (RR 0.08, 95% CI 0.01 to 0.55) and added tabs to facilitate doffing of masks (RR 0.33, 95%nCI 0.14 to 0.80) or gloves (RR 0.22, 95% CI 0.15 to 0.31). Guidance on PPE use: following the guidance and recommendations from the Centres for Disease Control and Prevention for doffing PPE compared to no guidance may reduce self-contamination (MD −5.44, 95% CI −7.43 to −3.45). One-step removal of gloves and gown compared to separate removal (RR 0.20, 95% CI 0.05 to 0.77), double gloving compared to single gloving (RR 0.34, 95% CI 0.17 to 0.66) and sanitising gloves before doffing with quaternary ammonium or bleach (but not alcohol-based hand rub) may decrease contamination. Additional verbal instructions may lead to fewer errors in doffing (MD −0.9, 95% CI −1.4 to −0.4). User training To a vast extent, face-to-face training may reduce non-compliance with doffing guidance (odds ratio 0.45, 95% CI 0.21 to 0.98) compared to solely providing folders or videos. In addition, computer simulation may lead to fewer errors in doffing (MD −1.2, 95% CI −1.6 to −0.7) and video lecture on donning PPE may lead to better skills scores (MD 30.70, 95% CI 20.14 to 41.26) compared to traditional lectures. Conclusions The more body parts are covered with PPE the better protection it offers. However, this is also associated and increased difficulty in donning and doffing PPE, and the PPE is less comfortable. Coveralls are the most difficult PPE to remove but may offer the best protection, followed by long gowns, gowns and aprons. The included studies had a high or unclear risk of bias, indirectness of evidence in simulation studies and small participant numbers. This increases the uncertainty about the estimates of effects, and it is likely that the true effects may be substantially different from the ones reported in this review.


title: Common cold decreases lung function in infants with recurrent wheezing

text: Abstract Background Common acute viral respiratory infections (colds) are the most frequent cause of exacerbations in infants with recurrent wheezing (RW). However, there is no quantitative information about the effect of colds on the lung function of infants with RW. This study was undertaken to determine the effect of common cold on forced expiratory parameters measured from raised lung volume in infants with RW. Methods Spirometric lung function (expiratory flows from raised lung volume) was randomly assessed in 28 infants with RW while they had a common cold and when asymptomatic. Results It was found that during colds there was a significant decrease in all forced expiratory parameters and this was much more evident for flows (FEF50%, FEF75% and FEF25–75%) which were definitively abnormal (less than −1.65 z-score) in the majority of infants. There was not association between family asthma, tobacco exposure, and other factors, with the extent of lung function decrease during colds. Tobacco during pregnancy but not a history of family asthma was significantly associated to lower expiratory flows; however, the association was significant only when infants were asymptomatic. Conclusion This study shows that common colds cause a marked reduction of lung function in infants with RW.


title: Mechanical Ventilation in an Airborne Epidemic

text: With the increasing threat of pandemic influenza and catastrophic bioterrorism, it is important for intensive care providers to be prepared to meet the challenge of large-scale airborne epidemics causing mass casualty respiratory failure. The severe acute respiratory syndrome outbreak exposed the vulnerability of health care workers and highlighted the importance of establishing stringent infection control and crisis management protocols. Patients who have acute lung injury and acute respiratory distress syndrome who require mechanical ventilation should receive a lung protective, low tidal volume strategy. Controversy remains regarding the use of high-frequency oscillatory ventilation and noninvasive positive pressure ventilation. Standard, contact, and airborne precautions should be instituted in intensive care units, with special care taken when aerosol-generating procedures are performed.


title: Protein Structure and Sequence Reanalysis of 2019-nCoV Genome Refutes Snakes as Its Intermediate Host and the Unique Similarity between Its Spike Protein Insertions and HIV-1

text: [Image: see text] As the infection of 2019-nCoV coronavirus is quickly developing into a global pneumonia epidemic, the careful analysis of its transmission and cellular mechanisms is sorely needed. In this Communication, we first analyzed two recent studies that concluded that snakes are the intermediate hosts of 2019-nCoV and that the 2019-nCoV spike protein insertions share a unique similarity to HIV-1. However, the reimplementation of the analyses, built on larger scale data sets using state-of-the-art bioinformatics methods and databases, presents clear evidence that rebuts these conclusions. Next, using metagenomic samples from Manis javanica, we assembled a draft genome of the 2019-nCoV-like coronavirus, which shows 73% coverage and 91% sequence identity to the 2019-nCoV genome. In particular, the alignments of the spike surface glycoprotein receptor binding domain revealed four times more variations in the bat coronavirus RaTG13 than in the Manis coronavirus compared with 2019-nCoV, suggesting the pangolin as a missing link in the transmission of 2019-nCoV from bats to human.


title: Using exercises to improve public health preparedness in Asia, the Middle East and Africa

text: BACKGROUND: Exercises are increasingly common tools used by the health sector and other sectors to evaluate their preparedness to respond to public health threats. Exercises provide an opportunity for multiple sectors to practice, test and evaluate their response to all types of public health emergencies. The information from these exercises can be used to refine and improve preparedness plans. There is a growing body of literature about the use of exercises among local, state and federal public health agencies in the United States. There is much less information about the use of exercises among public health agencies in other countries and the use of exercises that involve multiple countries. RESULTS: We developed and conducted 12 exercises (four sub-national, five national, three sub-regional) from August 2006 through December 2008. These 12 exercises included 558 participants (average 47) and 137 observers (average 11) from 14 countries. Participants consistently rated the overall quality of the exercises as very good or excellent. They rated the exercises lowest on their ability to identifying key gaps in performance. The vast majority of participants noted that they would use the information they gained at the exercise to improve their organization’s preparedness to respond to an influenza pandemic. Participants felt the exercises were particularly good at raising awareness and understanding about public health threats, assisting in evaluating plans and identifying priorities for improvement, and building relationships that strengthen preparedness and response across sectors and across countries. Participants left the exercises with specific ideas about the most important actions that they should engage in after the exercise such as improved planning coordination across sectors and countries and better training of health workers and response personnel. CONCLUSIONS: These experiences suggest that exercises can be a valuable, low-burden tool to improve emergency preparedness and response in countries around the world. They also demonstrate that countries can work together to develop and conduct successful exercises designed to improve regional preparedness to public health threats. The development of standardized evaluation methods for exercises may be an additional tool to help focus the actions to be taken as a result of the exercise and to improve future exercises. Exercises show great promise as tools to improve public health preparedness across sectors and countries.


title: Study of measles virus recombinant proteins and their immunobiological properties

text: Recombinant proteins rN (nucleocapsid) and rH/Nh (hemagglutinin) of the measles virus strain NovO/96 of genotype A were obtained. The immunobiological properties of the proteins were studied in the reaction with a panel of positive and negative sera. BALB/c mice were immunized with recombinant proteins and native antigen of the measles virus strain NovO/96 in order to obtain hyperimmune serum and its analysis using ELISA (enzyme-linked immunosorbent assay) and PRN (plaque reduction neutralization). The hyperimmune sera against recombinant proteins and native antigen of the measles virus strain NovO/96 were found to be highly active in ELISA. The antibodies against the proteins rN and rH/Nh were found to be capable of neutralizing the virus in titer 1 : 13.5 and 1 : 22.9, respectively. The neutralization titer of the antibodies generated against native antigen of the measles virus strain NovO/96 was 1 : 25.7.


title: Identification and characterization of Iflavirus 3C-like protease processing activities

text: Abstract Viral replication and capsid assembly in the viruses in the order Picornavirales requires polyprotein proteolytic processing by 3C or 3C-like (3CL) proteases. We identified and characterized the 3CL protease of Ectropis obliqua virus (EoV) of the newly established family Iflaviridae (order Picornavirales). The bacterially expressed EoV 3CL protease domain autocatalytically released itself from larger precursors by proteolytic cleavage, and cleavage sites were determined via N-terminal sequencing of the cleavage products. This protease also mediated trans-proteolytic activity and cleaved the polyprotein at the same specific positions. Moreover, we determined the critical catalytic residues (H2261, D2299, C2383) for the protease activity, and characterized the biochemical properties of EoV 3CL and its responses to various protease inhibitors. Our work is the first study to identify an iflaviral 3CL protease and further characterize it in detail and should foster our understanding of EoV and other iflaviruses.


title: Enteropathogens in pups from pet shops and breeding facilities

text: OBJECTIVES: To evaluate faecal and clinical scores and presence of several enteropathogens possibly implicated in the development of diarrhoea in pups aged between 6 and 16 weeks independently of their health status. METHODS: Pups were selected from pet shops and breeding facilities and assigned a faecal and clinical score. Standard isolation methods were used to determine presence of parasites, viruses and bacteria in faecal samples. For Escherichia coli, virulence genes were assessed by multiplex polymerase chain reaction. RESULTS: Fifty‐six pups were included in this study. Eighteen had no diarrhoea, 22 had no significant clinical signs related to gastroenteritis. Samples were positive for Toxocara canis (n=29), Giardia duodenalis (n=35), Cystoisospora (n=22), E. coli (n=47) and Clostridium perfringens (n=20). In four E. coli positive samples, genes were detected that correlate with pathogenicity in other animal species. A significant positive correlation between the presence of T. canis and faecal score was found. CLINICAL SIGNIFICANCE: Puppies obtained from a pet shop or breeding facility have a high risk of gastrointestinal disease. Furthermore, infectious agents may be present independently of faecal or clinical score. The identification of possible pathogenic E. coli strains suggests that their role in diarrhoea warrant further investigation.


title: Mental health nursing at a time of crisis

text: In Albert Camus' The Plague, the medical training of chief protagonist Dr Bernard Rieux fails to inoculate against his initial disbelief that an epidemic has arrived in the Algerian town of Oran. As Camus writes, 'Everybody knows that pestilences have a way of recurring in the world; yet somehow we find it hard to believe in ones that crash down on our heads from a blue sky' (Camus, 1947).


title: Decreased serum albumin level indicates poor prognosis of COVID-19 patients: hepatic injury analysis from 2,623 hospitalized cases

text: Coronavirus disease 2019 (COVID-19) is a global pandemic which has caused numerous deaths worldwide. The present study investigated the roles of hypoproteinemia in the clinical outcome and liver dysfunction of COVID-19 patients. In this retrospective study, we extracted data from 2,623 clinically confirmed adult COVID-19 patients (>18 years old) between January 29, 2020 and March 6, 2020 in Tongji Hospital, Wuhan, China. The patients were divided into three groups—non-critically ill, critically ill, and death groups—in accordance with the Chinese Clinical Guideline for COVID-19. Serum albumin, low-density lipoproteins cholesterol (LDL-C), and high-density lipoproteins cholesterol (HDL-C) concentrations and inflammatory cytokines levels were measured and compared among these three groups. The median age of these 2,623 patients was 64 years old (interquartile range (IQR), 52–71). Among the patients enrolled in the study, 2,008 (76.6%) were diagnosed as non-critically ill and 615 (23.4%) were critically ill patients, including 383 (14.6%) critically ill survivors and 232 (8.8%) critically ill deaths in the hospital. Marked hypoalbuminemia occurred in 38.2%, 71.2%, and 82.4% patients in non-critically ill, critically ill, and death groups, respectively, on admission and 45.9%, 77.7%, and 95.6% of these three groups, respectively, during hospitalization. We also discovered that serum low-density lipoprotein (LDL) and HDL levels were significantly lower in critically ill and death groups compared to non-critically ill group. Meanwhile, the patients displayed dramatically elevated levels of serum inflammatory factors, while a markedly prolonged activated partial thromboplastin time (APTT) in critically ill patients reflected coagulopathy. This study suggests that COVID-19-induced cytokine storm causes hepatotoxicity and subsequently critical hypoalbuminemia, which are associated with exacerbation of disease-associated inflammatory responses and progression of the disease and ultimately leads to death for some critically ill patients. ELECTRONIC SUPPLEMENTARY MATERIAL: Supplementary material is available for this article at 10.1007/s11427-020-1733-4 and is accessible for authorized users.


title: Comparative genomic analysis of pre-epidemic and epidemic Zika virus strains for virological factors potentially associated with the rapidly expanding epidemic

text: Less than 20 sporadic cases of human Zika virus (ZIKV) infection were reported in Africa and Asia before 2007, but large outbreaks involving up to 73% of the populations on the Pacific islands have started since 2007, and spread to the Americas in 2014. Moreover, the clinical manifestation of ZIKV infection has apparently changed, as evident by increasing reports of neurological complications, such as Guillain–Barré syndrome in adults and congenital anomalies in neonates. We comprehensively compared the genome sequences of pre-epidemic and epidemic ZIKV strains with complete genome or complete polyprotein sequences available in GenBank. Besides the reported phylogenetic clustering of the epidemic strains with the Asian lineage, we found that the topology of phylogenetic tree of all coding regions is the same except that of the non-structural 2B (NS2B) coding region. This finding was confirmed by bootscan analysis and multiple sequence alignment, which suggested the presence of a fragment of genetic recombination at NS2B with that of Spondweni virus. Moreover, the representative epidemic strain possesses one large bulge of nine bases instead of an external loop on the first stem-loop structure at the 3′-untranslated region just distal to the stop codon of the NS5 in the 1947 pre-epidemic prototype strain. Fifteen amino acid substitutions are found in the epidemic strains when compared with the pre-epidemic strains. As mutations in other flaviviruses can be associated with changes in virulence, replication efficiency, antigenic epitopes and host tropism, further studies would be important to ascertain the biological significance of these genomic changes.


COVID-19 and Interconnectedness


title: Synthesis and biological evaluation of phosphoramidate prodrugs of two analogues of 2-deoxy-d-ribose-1-phosphate directed to the discovery of two carbasugars as new potential anti-HIV leads

text: Abstract 2-Deoxy-α-d-ribose-1-phosphate is of great interest as it is involved in the biosynthesis and/or catabolic degradation of several nucleoside analogues of biological and therapeutic relevance. However due to the lack of a stabilising group at its 2-position, it is difficult to synthesize stable prodrugs of this compound. In order to overcome this lack of stability, the synthesis of carbasugar analogues of 2-deoxyribose-1-phosphate was envisioned. Herein the preparation of a series of prodrugs of two carbocyclic analogues of 2-deoxyribose-1-phosphate using the phosphoramidate ProTide technology, along with their biological evaluation against HIV and cancer cell proliferation, is reported.


title: Transient Oligomerization of the SARS-CoV N Protein – Implication for Virus Ribonucleoprotein Packaging

text: The nucleocapsid (N) phosphoprotein of the severe acute respiratory syndrome coronavirus (SARS-CoV) packages the viral genome into a helical ribonucleocapsid and plays a fundamental role during viral self-assembly. The N protein consists of two structural domains interspersed between intrinsically disordered regions and dimerizes through the C-terminal structural domain (CTD). A key activity of the protein is the ability to oligomerize during capsid formation by utilizing the dimer as a building block, but the structural and mechanistic bases of this activity are not well understood. By disulfide trapping technique we measured the amount of transient oligomers of N protein mutants with strategically located cysteine residues and showed that CTD acts as a primary transient oligomerization domain in solution. The data is consistent with the helical oligomer packing model of N protein observed in crystal. A systematic study of the oligomerization behavior revealed that altering the intermolecular electrostatic repulsion through changes in solution salt concentration or phosphorylation-mimicking mutations affects oligomerization propensity. We propose a biophysical mechanism where electrostatic repulsion acts as a switch to regulate N protein oligomerization.


title: Resilient and Agile Engineering Solutions to Address Societal Challenges such as Coronavirus Pandemic

text: The world is witnessing tumultuous times as major economic powers including the US, UK, Russia, India, and most of Europe continue to be in a state of lockdown. The worst-hit sectors due to this lockdown are sales, production (manufacturing), transport (aerospace and automotive) and tourism. Lockdowns became necessary as a preventive measure to avoid the spread of the contagious and infectious “Coronavirus Disease 2019” (COVID-19). This newly identified disease is caused by a new strain of the virus being referred to as Severe Acute Respiratory Syndrome CoronaVirus 2 (SARS CoV-2; formerly called 2019-nCoV). We review the current medical and manufacturing response to COVID-19, including advances in instrumentation, sensing, use of lasers, fumigation chambers and development of novel tools such as lab-on-the-chip using combinatorial additive and subtractive manufacturing techniques and use of molecular modelling and molecular docking in drug and vaccine discovery. We also offer perspectives on future considerations on climate change, outsourced versus indigenous manufacturing, automation, and antimicrobial resistance. Overall, this paper attempts to identify key areas where manufacturing can be employed to address societal challenges such as COVID-19.