title: Rescue of the 1947 Zika Virus Prototype Strain with a Cytomegalovirus Promoter-Driven cDNA Clone

text: The recent Zika virus (ZIKV) outbreak has been linked to severe pathogenesis. Here, we report the construction of a plasmid carrying a cytomegalovirus (CMV) promoter-expressed prototype 1947 Uganda MR766 ZIKV cDNA that can initiate infection following direct plasmid DNA transfection of mammalian cells. Incorporation of a synthetic intron in the nonstructural protein 1 (NS1) region of the ZIKV polyprotein reduced viral cDNA-associated toxicity in bacteria. High levels of infectious virus were produced following transfection of the plasmid bearing the wild-type MR766 ZIKV genome, but not one with a disruption to the viral nonstructural protein 5 (NS5) polymerase active site. Multicycle growth curve and plaque assay experiments indicated that the MR766 virus resulting from plasmid transfection exhibited growth characteristics that were more similar to its parental isolate than previously published 2010 Cambodia and 2015 Brazil cDNA-rescued ZIKV. This ZIKV infectious clone will be useful for investigating the genetic determinants of ZIKV infection and pathogenesis and should be amenable to construction of diverse infectious clones expressing reporter proteins and representing a range of ZIKV isolates. IMPORTANCE The study of ZIKV, which has become increasingly important with the recent association of this virus with microcephaly and Guillain-Barré syndrome, would benefit from an efficient strategy to genetically manipulate the virus. This work describes a model system to produce infectious virus in cell culture. We created a plasmid carrying the prototype 1947 Uganda MR766 ZIKV genome that both was stable in bacteria and could produce high levels of infectious virus in mammalian cells through direct delivery of this DNA. Furthermore, growth properties of this rescued virus closely resembled those of the viral isolate from which it was derived. This model system will provide a simple and effective means to study how ZIKV genetics impact viral replication and pathogenesis.


title: Connexin 43/47 channels are important for astrocyte/oligodendrocyte cross-talk in myelination and demyelination

text: The gap junctions (GJs), which form intercellular communicating channels between two apposing cells or form hemichannel with extracellular environment, perform crucial functions to maintain small molecule homeostasis. The central nervous system (CNS) GJs are important for maintenance of myelin sheath and neuronal activity. Connexin (Cx) proteins are building blocks of GJs. Recent cell-biological investigations show that amongst the CNS specific Cxs, the most abundant Cx protein, Cx43 and its oligodendrocytic coupling partner Cx47 primarily important for maintenance of CNS myelin. Recent investigations elucidate that the expression of Cx43 and Cx47 is very important to maintain K+ buffering and nutrient homeostasis in oligodendrocytes, CNS myelin and oligodendrocyte function. The investigations on Multiple Sclerosis (MS) patient samples and EAE hypothesized that the functional loss of Cx43/Cx47 could be associated with spread of chronic MS lesions. Exploring the mechanism of initial GJ alteration and its effect on demyelination in this model of MS might play a primary role to understand the basis of altered CNS homeostasis, observed during MS. In this review, we mainly discuss the role of CNS GJs, specifically the Cx43/Cx47 axis in the perspective of demyelination.


title: Protective effect of intranasal peste des petits ruminants virus and bacterin vaccinations: Clinical, hematological, serological, and serum oxidative stress changes in challenged goats

text: BACKGROUND AND AIM: The current vaccination for peste des petits ruminants virus (PPRV) is stalled by myriad challenges and continuous endemicity of pneumonia due to fulminant bacterial complication in goats. The present study evaluated the protective effect of intranasal PPRV linage 1 and bacterine vaccinations. MATERIALS AND METHODS: Twelve West African Dwarf (WAD) goats aged 6 months were randomly grouped and vaccinated within 2 weeks using a combination of PPRV lineage 1 vaccine (Nig/75), and bacterin from Mannheimia haemolytica (Mh) or Pasteurella multocida intranasally. The goats were observed for 3 weeks post-vaccination before comingled with a known infected WAD goat with apparent clinical signs of peste des petits ruminants and further observed clinically for 5 weeks post-infection (PI). Blood samples were taken for hematology while sera were assayed for antioxidants (glutathione peroxidase, glutathione transferase, and superoxide dismutase) activities and pro-oxidants (malondialdehyde content, reduced glutathione, hydrogen peroxide generation(,) and myeloperoxidase) using spectrophotometric methods. Data were subjected to parametric statistics at α=0.05 using GraphPad Prism version 21. RESULTS: Clinically, there were pyrexia, oculonasal discharge, diarrhea, anemia, leukopenia, and increased pro-oxidants in the unvaccinated goats, while moderate neutrophilia and leukocytosis were observed in PPRV and bacterin vaccinated goats. Two unvaccinated goats were weak and euthanized at 13 and 28 days PI. The goats vaccinated with PPRV and Mh showed better response clinically and biochemically. CONCLUSION: The mucosal vaccination of goats with PPRV vaccine and bacterine will protect against exposure and culminate in the development of protective mucosal, humoral, and cell-mediated immune responses. This vaccination strategy will provide framework needed in the prevention and control of endemic caprine pneumonia in Nigeria.


title: Activation of protein kinase R is required for induction of stress granules by respiratory syncytial virus but dispensable for viral replication

text: Abstract We performed experiments to determine the effect of PKR activation on respiratory syncytial virus (RSV) replication. We first determined that RSV infection activates PKR which induces the phosphorylation of eIF2α, resulting in the formation of host stress granules. We used RNA interference to decrease endogenous PKR levels. RSV replication was not altered in cells deficient for PKR expression. However, RSV-mediated stress granule formation was significantly reduced in PKR-knockdown cells. As an alternative method to block PKR activation, we used treatment with the kinase inhibitor 2-aminopurine (2-AP). We observed that 2-AP treatment significantly reduced viral replication. We also treated PKR-knockdown cells with 2-AP and inoculated with RSV. Under these conditions, 2-AP treatment diminished viral replication in the absence of PKR expression. These results suggest that PKR activation has a minimal effect on RSV replication and that the antiviral effect of 2-AP during RSV infection likely occurs via a PKR-independent mechanism.


title: Risk assessment and laboratory investigation of respiratory illness in travellers returning to Singapore 2012–2015: experience from the MERS-CoV Surveillance Programme

text: Since the emergence of Middle East respiratory syndrome coronavirus (MERS-CoV), Singapore has enhanced its national surveillance system to detect the potential importation of this novel pathogen. Using the guidelines from the Singapore Ministry of Health, a suspect case was defined as a person with clinical signs and symptoms suggestive of pneumonia or severe respiratory infection with breathlessness, and with an epidemiological link to countries where MERS-CoV cases had been reported within the preceding 14 days. This report describes a retrospective review of 851 suspected MERS-CoV cases assessed at the adult tertiary-care hospital in Singapore between September 2012 and December 2015. In total, 262 patients (31%) were hospitalized. All had MERS-CoV infection ruled out by RT–PCR or clinical assessment. Two hundred and thirty (88%) of the hospitalized patients were also investigated for influenza virus by RT–PCR. Of these, 62 (27%) tested positive for seasonal influenza. None of the patients with positive influenza results had been vaccinated in the year prior to hospital admission. Ninety-three (36%) out of the 262 hospitalized patients had clinical and/or radiological evidence of pneumonia. This study demonstrates the potential benefits of pre-travel vaccination against influenza and pneumococcal disease.


COVID-19 exposes animal agriculture’s vulnerability


title: Current Trends in Diagnostics of Viral Infections of Unknown Etiology

text: Viruses are evolving at an alarming rate, spreading and inconspicuously adapting to cutting-edge therapies. Therefore, the search for rapid, informative and reliable diagnostic methods is becoming urgent as ever. Conventional clinical tests (PCR, serology, etc.) are being continually optimized, yet provide very limited data. Could high throughput sequencing (HTS) become the future gold standard in molecular diagnostics of viral infections? Compared to conventional clinical tests, HTS is universal and more precise at profiling pathogens. Nevertheless, it has not yet been widely accepted as a diagnostic tool, owing primarily to its high cost and the complexity of sample preparation and data analysis. Those obstacles must be tackled to integrate HTS into daily clinical practice. For this, three objectives are to be achieved: (1) designing and assessing universal protocols for library preparation, (2) assembling purpose-specific pipelines, and (3) building computational infrastructure to suit the needs and financial abilities of modern healthcare centers. Data harvested with HTS could not only augment diagnostics and help to choose the correct therapy, but also facilitate research in epidemiology, genetics and virology. This information, in turn, could significantly aid clinicians in battling viral infections.


title: Electron tomography in life science

text: Abstract Electron tomography (ET) is a three-dimensional technique suitable to study pleomorphic biological structures with nanometer resolution. This makes the methodology remarkably versatile, allowing the exploration of a large range of biological specimens, both in an isolated state and in their cellular context. The application of ET has undergone an exponential growth over the last decade, enabled by seminal technological advances in methods and instrumentation, and is starting to make a significant impact on our understanding of the cellular world. While the attained results are already remarkable, ET remains a young technique with ample potential to be exploited. Current developments towards large-scale automation, higher resolution, macromolecular labeling and integration with other imaging techniques hold promise for a near future in which ET will extend its role as a pivotal tool in structural and cell biology.


Should we try the antiinflammatory natural product, celastrol, for COVID‐19?


title: Clinical and Molecular Features of Feline Foamy Virus and Feline Leukemia Virus Co-Infection in Naturally-Infected Cats

text: Feline foamy virus (FFV) and feline leukemia virus (FeLV) belong to the Retroviridae family. While disease has not been reported for FFV infection, FeLV infection can cause anemia and immunosuppression (progressive infection). Co-infection with FFV/FeLV allows evaluation of the pathogenic potential and epidemiology of FFV infection in cats with FeLV pathology. Blood and buccal swab samples from 81 cats were collected in Rio de Janeiro. Plasma was serologically tested for FeLV. DNA extracted from peripheral blood mononuclear cells and buccal swabs was used to PCR detect FFV and FeLV. A qPCR was developed to detect and measure FFV proviral loads (pVLs) in cats. FeLV qPCR was performed using previous methods. The median log10 pVL of FFV mono-infected individuals was lower than found in FFV/FeLV co-infected cats in buccal swabs (p = 0.003). We found 78% of cats had detectable buccal FFV DNA in FFV mono-infected and FFV co-infected FeLV-progressive cats, while in FeLV-regressive cats (those without signs of disease) 22% of cats had detectable buccal FFV DNA (p = 0.004). Our results suggest that regressive FeLV infection may reduce FFV saliva transmission, the main mode of FV transmission. We did not find evidence of differences in pathogenicity in FFV mono- and -dually infected cats. In summary, we show that FVs may interact with FeLV within the same host. Our study supports the utility of cats naturally co-infected with retroviruses as a model to investigate the impact of FV on immunocompromised mammalian hosts.


title: The cauliflower mosaic virus reverse transcriptase is not produced by the mechanism of ribosomal frameshifting in Saccharomyces cerevisiae

text: Abstract The capsid protein and the reverse transcriptase of cauliflower mosaic virus (CaMV) are encoded by two genes (ORF IV and ORF V) that lie in different translation reading frames. A comparison can be drawn between the synthesis of both CaMV proteins and the fusion protein in a yeast retrotransposon, Ty, resulting from a +1 frameshifting event which fuses two out-of-phase ORFs encoding the structural protein and the reverse transcriptase of Ty. For this reason, we constructed a yeast expression vector containing CaMV ORF VII fused to CaMV ORF III by a fragment of 452 bp including the overlapping region of ORF IV and ORF V, ORF VII and ORF III being used as reporter genes. We characterized two proteins (22 and 50 kDa) synthesized from this plasmid in the yeast expression system. We demonstrated that the 50-kDa polypeptide is not synthesized from a +1 frameshifting event but is probably a dimeric form of the 22-kDa protein. From this result we conclude that the CaMV reverse transcriptase is not produced by a mechanism of ribosomal frameshifting.


The Author Reply: A Comment on “Quaternary Prevention in Public Health”


title: Multi-perspective comparisons and mitigation implications of SO(2) and NO(x) discharges from the industrial sector of China: a decomposition analysis

text: This study is the first attempt to investigate the drivers of Chinese industrial SO(2) and NO(x) emissions from both periodic and structural perspectives through a decomposition analysis using the logarithmic mean Divisia index (LMDI). The two pollutants’ emissions were decomposed into output effects, structural effects, clean production effects, and pollution abatement effects. The results showed that China’s industrial SO(2) discharge increased by 1.14 Mt during 2003–2014, and the contributions from the four effects were 23.17, − 1.88, − 3.80, and − 16.36 Mt, respectively. Likewise, NO(x) discharge changed by − 3.44 Mt over 2011–2014, and the corresponding contributions from the four effects were 2.97, − 0.62, − 1.84, and − 3.95 Mt. Thus, the output effect was mainly responsible for the growth of the two discharges. The average annual contribution rates of SO(2) and NO(x) from output were 14.33 and 5.97%, respectively, but pollution abatement technology presented the most obvious mitigating effects (− 10.11 and − 7.92%), followed by the mitigating effects of clean production technology (− 2.35 and − 3.7%), and the mitigation from the structural effect was the weakest (− 1.16 and − 1.25%, respectively), which meant pollutant reduction policies related to industrial structure adjustment should be a long-term measure for the two discharges. In addition, the sub-sectors of I20 (manufacture of raw chemical materials and chemical products), I24 (manufacture of non-metallic mineral products), and I26 (smelting and pressing of non-ferrous metals) were the major contributors to both discharges. Thus, these sub-sectors should be given priority consideration when designing mitigation-related measures. Last, some particular policy implications were recommended for reducing the two discharges, including that the government should seek a technological discharge reduction route. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s11356-018-1306-x) contains supplementary material, which is available to authorized users.


title: Targeting endosomal acidification by chloroquine analogs as a promising strategy for the treatment of emerging viral diseases

text: Emerging viruses such as HIV, dengue, influenza A, SARS coronavirus, Ebola, and other viruses pose a significant threat to human health. Majority of these viruses are responsible for the outbreaks of pathogenic lethal infections. To date, there are no effective therapeutic strategies available for the prophylaxis and treatment of these infections. Chloroquine analogs have been used for decades as the primary and most successful drugs against malaria. Concomitant with the emergence of chloroquine‐resistant Plasmodium strains and a subsequent decrease in the use as antimalarial drugs, other applications of the analogs have been investigated. Since the analogs have interesting biochemical properties, these drugs are found to be effective against a wide variety of viral infections. As antiviral action, the analogs have been shown to inhibit acidification of endosome during the events of replication and infection. Moreover, immunomodulatory effects of analogs have been beneficial to patients with severe inflammatory complications of several viral diseases. Interestingly, one of the successful targeting strategies is the inhibition of HIV replication by the analogs in vitro which are being tested in several clinical trials. This review focuses on the potentialities of chloroquine analogs for the treatment of endosomal low pH dependent emerging viral diseases.


title: Rapid semi-automated quantitative multiplex tandem PCR (MT-PCR) assays for the differential diagnosis of influenza-like illness

text: BACKGROUND: Influenza A, including avian influenza, is a major public health threat in developed and developing countries. Rapid and accurate detection is a key component of strategies to contain spread of infection, and the efficient diagnosis of influenza-like-illness is essential to protect health infrastructure in the event of a major influenza outbreak. METHODS: We developed a multiplexed PCR (MT-PCR) assay for the simultaneous diagnosis of respiratory viruses causing influenza-like illness, including the specific recognition of influenza A haemagglutinin subtypes H1, H3, and H5. We tested several hundred clinical specimens in two diagnostic reference laboratories and compared the results with standard techniques. RESULTS: The sensitivity and specificity of these assays was higher than individual assays based on direct antigen detection and standard PCR against a range of control templates and in several hundred clinical specimens. The MT-PCR assays provided differential diagnoses as well as potentially useful quantitation of virus in clinical samples. CONCLUSIONS: MT-PCR is a potentially powerful tool for the differential diagnosis of influenza-like illness in the clinical diagnostic laboratory.


title: Predicting optimal lockdown period with parametric approach using three-phase maturation SIRD model for COVID-19 pandemic

text: This paper proposes a three-phase Susceptible-Infected-Recovered-Dead (SIRD) model to calculate an optimal lockdown period for some specific geographical regions that will be favorable to break not only the transmission chain but also will help country’s economy to recover and support infrastructure in a fight against COVID-19. Proposed model is novel since it additionally includes parameters i.e. silent carriers, sociability of newly infected person and unregistered died coronavirus infected people along with the infection rate, suspected rate and death rate. These parameters contribute a lot to figure out the more clear model, along with essential parameters. The model takes the testing rate of suspected people into consideration and this rate varies with respect to phase of the epidemic growth. Proposed SIRD model is divided into three phases based on the awareness and sustainability of disease. Time is divided into different periods as rate of infection and recovery fluctuates region to region. The model is tested on China data and is efficient enough to propose a model very close to their actual figures of infected people, recovered people, died and active cases. The model predicts the optimal lockdown period as 73 days for China which is very close to their actual lockdown period (77 days). Further, the model is implemented to predict the optimal lockdown period of India and Italy.


title: CARES Act to help providers bolster care; supports communities, businesses

text: As the country continues to grapple with this unprecedented COVID‐19 pandemic, mental health field leaders are responding to what the most recent $2 trillion stimulus aid package passed by the Senate and House and signed by President Trump means for mental health and substance use providers.


title: Chapter 9 Trends in antiviral strategies

text: Abstract Viral populations are true moving targets regarding the genomic sequences to be targeted in antiviral designs. Experts from different fields have expressed the need for new paradigms for antiviral interventions and viral disease control. This chapter reviews several antiviral strategies that aim at counteracting the adaptive capacity of viral quasispecies. The proposed designs are based on combinations of different antiviral drugs and immune modulators, or in the administration of virus-specific mutagenic agents, in an approach termed lethal mutagenesis of viruses. It consists of decreasing viral fitness by an excess of mutations that render viral proteins suboptimal or nonfunctional. Viral extinction by lethal mutagenesis involves several sequential, overlapping steps that recapitulate the major concepts of intrapopulation interactions and genetic information stability discussed in preceding chapters. Despite the magnitude of the challenge, the chapter closes with some optimistic prospects for effective control of viruses displaying error-prone replication, based on the combined targeting of replication fidelity and the induction of the innate immune response.


title: Complementary and Integrative Treatments Upper Respiratory Infection

text: The combination of nasal congestion, rhinorrhea, sore throat, cough, and malaise is the symptomatic profile that constitutes an uncomplicated upper respiratory tract infection (URI), also known as the common cold. Because no known cure exists for a URI, numerous products are available, each marketed with the promise of alleviating the associated symptoms and/or shortening the duration of illness. The evidence supporting these claims is variable and is the focus of this article, with an emphasis on complementary and integrative therapies.


title: Prevalence of SARS-CoV-2 infection in previously undiagnosed health care workers at the onset of the U.S. COVID-19 epidemic

text: IMPORTANCE: Healthcare workers are presumed to be at increased risk of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection due to occupational exposure to infected patients. However, no epidemiological study has examined the prevalence of SARS-CoV-2 infection in a cohort of healthcare workers during the early phase of community transmission. OBJECTIVE: To determine the baseline prevalence of SARS-CoV-2 infection in a cohort of previously undiagnosed healthcare workers and a comparison group of non-healthcare workers. DESIGN: Prospective cohort study SETTING: A large U.S. university and two affiliated university hospitals PARTICIPANTS: 546 health care workers and 283 non-health care workers with no known prior SARS-CoV-2 infection EXPOSURE: Healthcare worker status and role MAIN OUTCOME(S) AND MEASURE(S): SARS-CoV-2 infection status as determined by presence of SARS-CoV-2 RNA in oropharyngeal swabs. RESULTS: At baseline, 41 (5.0%) of participants tested positive for SARS-CoV-2 infection, of whom 14 (34.2%) reported symptoms. The prevalence of SARS-CoV-2 infection was higher among healthcare workers (7.3%) than in non-healthcare workers (0.4%), representing a 7.0% greater absolute risk (95% confidence interval for risk difference 4.7%, 9.3%). The majority of infected healthcare workers (62.5%) worked as nurses. Positive tests increased across the two weeks of cohort recruitment in line with rising confirmed cases in the hospitals and surrounding counties. CONCLUSIONS AND RELEVANCE: In a prospective cohort conducted in the early phases of community transmission, healthcare workers had a higher prevalence of SARS-CoV-2 infection than non-healthcare workers, attesting to the occupational hazards of caring for patients in this crisis. Baseline data reported here will enable us to monitor the spread of infection and examine risk factors for transmission among healthcare workers. These results will inform optimal strategies for protecting the healthcare workforce, their families, and their patients. CLINICALTRIALS.GOV REGISTRATION NUMBER: NCT04336215


title: Infection prevention and control competencies for hospital-based health care personnel

text: BACKGROUND: Infection prevention and control education for hospital-based health care personnel has differed across organizations because of a lack of identified practice competencies. This gap also has resulted in variation of the educational curriculum in the academic setting and a lack of consistent preparation for emergency responses. The purpose of this study was to develop a list of competencies and measurable activities, or terminal objectives, for hospital-based health care personnel applicable for use during routine patient care activities as well as during natural and man-made disasters. METHODS: Competencies and terminal objectives related to infection prevention and control were developed using an evidence-based approach comprising the following steps: (a) review of the literature, (b) review of existing competencies and published curricula/training objectives, (c) synthesis of new competencies and terminal objectives, (d) expert panel review and competency refinement using the Delphi survey process, and (e) delineation of competencies by occupation. The 8 disciplines addressed were nurses (RNs and LPNs), nursing assistants, physicians, respiratory therapists, physical and occupational therapists, environmental services, laboratory professionals, and ancillary staff. RESULTS: An initial list of competency statements and terminal objectives were compiled and then vetted by a Delphi panel of experts in infection prevention and control until > 80% agreement was achieved on all competency statements and terminal objectives. CONCLUSION: The final matrix of competencies and terminal objectives developed through this process may be used as a content framework for educational curricula and training materials for hospital-based health care personnel. The process also may be of use in determining the core competencies and terminal objectives regarding infection prevention and control for health care personnel in other settings. Validation of these results is an important next step.


title: Mice with a human touch

text: Years of tinkering with the mouse immune system genes has finally produced a winner, with the approval of the first fully human monoclonal antibody made in a mouse. A real breakthrough or an incremental improvement? Christopher Thomas Scott investigates.


title: A nonparametric estimation of the infection curve

text: Predicting the future course of an epidemic depends on being able to estimate the current numbers of infected individuals. However, while back-projection techniques allow reliable estimation of the numbers of infected individuals in the more distant past, they are less reliable in the recent past. We propose two new nonparametric methods to estimate the unobserved numbers of infected individuals in the recent past in an epidemic. The proposed methods are noniterative, easily computed and asymptotically normal with simple variance formulas. Simulations show that the proposed methods are much more robust and accurate than the existing back projection method, especially for the recent past, which is our primary interest. We apply the proposed methods to the 2003 Severe Acute Respiratory Syndorme (SARS) epidemic in Hong Kong.


title: High-affinity Na+-dependent dicarboxylate cotransporter promotes cellular senescence by inhibiting SIRT1

text: Abstract High-affinity Na+-dependent dicarboxylate cotransporter (NaDC3) can transport Krebs cycle intermediates into cells. Our previous study has shown that NaDC3 promotes cellular senescence, but its mechanism is not clear. It is known that when the concentration of intermediates in Krebs cycle is increased, NAD+/NADH ratio will be decreased. NAD+-dependent histone deacetylase sirtuin1 (SIRT1) prolongs mammalian cellular lifespan. Therefore, we propose that NaDC3 accelerates cellular aging by inhibiting SIRT1. After NaDC3 was overexpressed in two human embryo lung fibroblastic cell lines, WI38 and MRC-5, we found that the cells displayed aging-related phenotypes in advance. Meanwhile, the level of SIRT1 activity was down-regulated. In WI38/hNaDC3 cells treated with the activators of SIRT1, aging-related phenotypes induced by NaDC3 were obviously improved. The NAD+/NADH ratio in WI38/hNaDC3 cells was also decreased. Further study found that enhanced intracellular NAD+ level could attenuate the aging phenotypes induced by NaDC3. Thus, NaDC3 promotes cellular senescence probably by inhibiting NAD+-dependent SIRT1.


Virus linked with prostate cancer


title: Comparative analysis reveals adaptive evolution of bat IFITMs and a novel antiviral determinant

text: Host interferon-induced transmembrane proteins (IFITMs) are broad-spectrum antiviral restriction factors. Of these, IFITM3 potently inhibits viruses that enter cells through acidic endosomes, many of which are zoonotic and emerging viruses with bats (order Chiroptera) as natural hosts. We previously demonstrated that microbat IFITM3 is antiviral. Here we show that bat IFITMs are characterized by strong adaptive evolution and identify a highly variable and functionally important site - codon 70 - within the conserved CD225 domain of IFITMs. Mutation of this residue in microbat IFITM3 impairs restriction of four different virus families that enter cells via endosomes. This mutant shows altered subcellular localization and reduced S-palmitoylation, a phenotype copied by mutation of conserved cysteine residues in microbat IFITM3. Furthermore, we show that microbat IFITM3 is S-palmitoylated on cysteine residues C71, C72 and C105, mutation of each cysteine residue individually impairs virus restriction, and a triple C71-C72-C105 mutant loses all restriction, concomitant with subcellular re-localization of microbat IFITM3 to Golgi-associated sites. Thus, we propose that S-palmitoylation is critical for Chiropteran IFITM3 function and identify a key molecular determinant of IFITM3 S-palmitoylation.


title: Synthetic and semi-synthetic strategies to study ubiquitin signaling

text: The post-translational modification ubiquitin can be attached to the ɛ-amino group of lysine residues or to a protein's N-terminus as a mono ubiquitin moiety. Via its seven intrinsic lysine residues and its N-terminus, it can also form ubiquitin chains on substrates in many possible ways. To study ubiquitin signals, many synthetic and semi-synthetic routes have been developed for generation of ubiquitin-derived tools and conjugates. The strength of these methods lies in their ability to introduce chemo-selective ligation handles at sites that currently cannot be enzymatically modified. Here, we review the different synthetic and semi-synthetic methods available for ubiquitin conjugate synthesis and their contribution to how they have helped investigating conformational diversity of diubiquitin signals. Next, we discuss how these methods help understanding the ubiquitin conjugation–deconjugation system by recent advances in ubiquitin ligase probes and diubiquitin-based DUB probes. Lastly, we discuss how these methods help studying post-translational modification of ubiquitin itself.


title: Methods of inactivation of SARS-CoV-2 for downstream biological assays

text: The scientific community has responded to the COVID-19 pandemic by rapidly undertaking research to find effective strategies to reduce the burden of this disease. Encouragingly, researchers from a diverse array of fields are collectively working towards this goal. Research with infectious SARS-CoV-2 is undertaken in high containment laboratories, however, it is often desirable to work with samples at lower containment levels. To facilitate the transfer of infectious samples from high containment laboratories, we have tested methods commonly used to inactivate virus and prepare the sample for additional experiments. Incubation at 80°C, and a range of detergents and UV energies were successful at inactivating a high titre of SARS-CoV-2. These protocols can provide a framework for in house inactivation of SARS-CoV-2 in other laboratories, ensuring the safe use of samples in lower containment levels.


title: Porcine epidemic diarrhea virus does not replicate in porcine monocyte-derived dendritic cells, but activates the transcription of type I interferon and chemokine

text: Abstract Porcine epidemic diarrhea virus (PEDV) belongs to the alphacoronavirus of the Coronaviridae. It is the major etiological agent of the recent outbreaks of piglet diarrhea and death in the US. Limited knowledge is currently available regarding the role of dendritic cells in PEDV infection. Here, we observed that PEDV did not replicate in monocyte-derived dendritic cells as evidenced by the decrease of viral gene transcript copies in infected cells by qRT-PCR and the absence of viral proteins by immunofluorescence staining as well as the absence of virus particles in infected cells by transmission electron microscopy. In addition, PEDV did not compromise cell viability at 48, 72, and 96h after infection at either a MOI of 2.5 or 5. Interestingly, an increased transcription of type I interferon including interferon-α and β was observed in infected cells compared to mock infected cells. Surprisingly, we did not detect any interferon-β in the supernatants of infected cells. A slight increase in interferon-α protein production in the supernatants of PEDV-infected cells was observed compared to mock infected cells. We also observed a markedly increased transcription of interferon inducible protein −10 (IP-10). Overall, PEDV does not replicate in porcine Mo-DC, but activates the transcription of type I interferon and chemokine IP-10.


3D PRINTING OF FACE SHIELDS DURING COVID-19 PANDEMIC: A TECHNICAL NOTE


title: Borna Disease Virus and the Brain

text: Abstract Viruses with the ability to establish persistent infection in the central nervous system (CNS) can induce progressive neurologic disorders associated with diverse pathological manifestations. Clinical, epidemiological, and virological evidence supports the hypothesis that viruses contribute to human mental diseases whose etiology remains elusive. Therefore, the investigation of the mechanisms whereby viruses persist in the CNS and disturb normal brain function represents an area of research relevant to clinical and basic neurosciences. Borna disease virus (BDV) causes CNS disease in several vertebrate species characterized by behavioral abnormalities. Based on its unique features, BDV represents the prototype of a new virus family. BDV provides an important model for the investigation of the mechanisms and consequences of viral persistence in the CNS. The BDV paradigm is amenable to study virus–cell interactions in the CNS that can lead to neurodevelopmental abnormalities, immune-mediated damage, as well as alterations in cell differentiated functions that affect brain homeostasis. Moreover, seroepidemiological data and recent molecular studies indicate that BDV is associated with certain neuropsychiatric diseases. The potential role of BDV and of other yet to be uncovered BDV-related viruses in human mental health provides additional impetus for the investigation of this novel neurotropic infectious agent.


title: Decoding the enigma of antiviral crisis: Does one target molecule regulate all?

text: Abstract Disease fatality associated with Ebola, SARS-CoV and dengue infections in humans is attributed to a cytokine storm that is triggered by excessive pro-inflammatory responses. Interleukin (IL)-6 acts as a mediator between pro- and anti-inflammatory reactivity by initiating trans- and classical-signaling, respectively. Hence, IL-6 is assumed to provide a target for a broad range of antiviral agents. Available immunosuppressive antivirals are directed to control an often exaggerated pro-inflammatory response that gives rise to complex clinical conditions such as lymphocytopenia. It is known that IL-6, via its soluble receptor (sIL-6R), initiates a pro-inflammatory response while an anti-inflammatory response is triggered by the membrane-bound IL-6 receptor (IL-6R). Future antivirals should thus aim to target the mechanism that regulates switching between IL-6 trans- and classical-signaling. In this review, we propose that the tumour necrosis factor-α converting enzyme ADAM-17 could be the master molecule involved in regulating IL-6 class switching and through this in controlling pro- and anti-inflammatory responses to viral antigenic stimuli. Therefore, ADAM-17 should be considered as a potential target molecule for novel antiviral drug discovery that would regulate host reactivity to infection and thereby limit or prevent fatal outcomes.


July 2020 New in Review


title: Properties of Two Enterovirus Antibodies that are Utilized in Diabetes Research

text: Human enteroviruses (EVs) comprise >100 different types. Research suggests a non-chance association between EV infections and type 1 diabetes. Immunohistochemical studies with the anti-EV antibody 5D-8.1 have shown that the EV capsid antigen is present in pancreatic islet cells of diabetic subjects. When it was noticed that 5D-8.1 may cross-react with human proteins, doubt was casted on the significance of the above histopathologic findings. To address this issue, properties of EV antibodies 5D-8.1 and 9D5 have been investigated using peptide microarrays, peptide substitution scanning, immunofluorescence of EV-infected cells, EV neutralization assays, bioinformatics analysis. Evidence indicates that the two antibodies bind to distinct non-neutralizing linear epitopes in VP1 and are specific for a vast spectrum of EV types (not for other human viruses). However, their epitopes may align with a few human proteins at low expected values. When tested by immunofluorescence, high concentrations of 5D-8.1 yelded faint cytoplasmic staining in uninfected cells. At reduced concentrations, both antibodies produced dotted staining only in the cytoplasm of infected cells and recognized both acute and persistent EV infection. Thus, the two monoclonals represent distinct and independent probes for hunting EVs in tissues of patients with diabetes or other endocrine conditions.


title: Houttuynia cordata Thunb. and its bioactive compound 2-undecanone significantly suppress benzo(a)pyrene-induced lung tumorigenesis by activating the Nrf2-HO-1/NQO-1 signaling pathway

text: BACKGROUND: Lung cancer remains the most common cause of cancer-related deaths, with a high incidence and mortality in both sexes worldwide. Chemoprevention has been the most effective strategy for lung cancer prevention. Thus, exploring novel and effective candidate agents with low toxicity for chemoprevention is essential and urgent. Houttuynia cordata Thunb. (Saururaceae) (H. cordata), which is a widely used herbal medicine and is also popularly consumed as a healthy vegetable, exhibits anti-inflammatory, antioxidant and antitumor activity. However, the chemopreventive effect of H. cordata against benzo(a)pyrene (B[a]P)-initiated lung tumorigenesis and the underlying mechanism remain unclear. METHODS: A B[a]P-stimulated lung adenocarcinoma animal model in A/J mice in vivo and a normal lung cell model (BEAS.2B) in vitro were established to investigate the chemopreventive effects of H. cordata and its bioactive compound 2-undecanone against lung tumorigenesis and to clarify the underlying mechanisms. RESULTS: H. cordata and 2-undecanone significantly suppressed B[a]P-induced lung tumorigenesis without causing obvious systemic toxicity in mice in vivo. Moreover, H. cordata and 2-undecanone effectively decreased B[a]P-induced intracellular reactive oxygen species (ROS) overproduction and further notably protected BEAS.2B cells from B[a]P-induced DNA damage and inflammation by significantly inhibiting phosphorylated H2A.X overexpression and interleukin-1β secretion. In addition, H. cordata and 2-undecanone markedly activated the Nrf2 pathway to induce the expression of the antioxidative enzymes heme oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO-1). Nrf2 silencing by transfection with Nrf2 siRNA markedly decreased the expression of HO-1 and NQO-1 to diminish the reductions in B[a]P-induced ROS overproduction, DNA damage and inflammation mediated by H. cordata and 2-undecanone. CONCLUSIONS: H. cordata and 2-undecanone could effectively activate the Nrf2-HO-1/NQO-1 signaling pathway to counteract intracellular ROS generation, thereby attenuating DNA damage and inflammation induced by B[a]P stimulation and playing a role in the chemoprevention of B[a]P-induced lung tumorigenesis. These findings provide new insight into the pharmacological action of H. cordata and indicate that H. cordata is a novel candidate agent for the chemoprevention of lung cancer. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13046-019-1255-3) contains supplementary material, which is available to authorized users.


title: Clinical characteristics of COVID-19 infection in chronic obstructive pulmonary disease: a multicenter, retrospective, observational study.

text: Background Coronavirus disease 2019 (COVID-19) has been a global pandemic disease, with more than 4 million cases and nearly 300,000 deaths. Little is known about COVID-19 in patients with chronic obstructive pulmonary disease (COPD). We aimed to evaluate the influence of preexisting COPD on the progress and outcomes of COVID-19. Methods This was a multicenter, retrospective, observational study. We enrolled 1,048 patients aged 40 years and above, including 50 patients with COPD and 998 patients without COPD, and with COVID-19 confirmed via high-throughput sequencing or real-time reverse transcription-polymerase chain reaction, between December 11, 2019 and February 20, 2020. We collected data of demographics, pathologic test results, radiologic imaging, and treatments. The primary outcomes were composite endpoints determined by admission to an intensive care unit, the use of mechanical ventilation, or death. Results Compared with patients who had COVID-19 but not COPD, those with COPD had higher rates of fatigue (56.0% vs. 40.2%), dyspnea (66.0% vs. 26.3%), diarrhea (16.0% vs. 3.6%), and unconsciousness (8.0% vs. 1.7%) and a significantly higher proportion of increased activated partial thromboplastin time (23.5% vs. 5.2%) and D-dimer (65.9% vs. 29.3%), as well as ground-glass opacities (77.6% vs. 60.3%), local patchy shadowing (61.2% vs. 41.4%), and interstitial abnormalities (51.0% vs. 19.8%) on chest computed tomography. Patients with COPD were more likely to develop bacterial or fungal coinfection (20.0% vs. 5.9%), acute respiratory distress syndrome (ARDS) (20.0% vs. 7.3%), septic shock (14.0% vs. 2.3%), or acute renal failure (12.0% vs. 1.3%). Patients with COPD and COVID-19 had a higher risk of reaching the composite endpoints [hazard ratio (HR): 2.17, 95% confidence interval (CI): 1.40-3.38; P=0.001] or death (HR: 2.28, 95% CI: 1.15-4.51; P=0.019), after adjustment. Conclusions In this study, patients with COPD who developed COVID-19 showed a higher risk of admission to the intensive care unit, mechanical ventilation, or death.


title: Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay

text: Abstract Objective To evaluate a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of SARS-CoV-2, and compare it with RT polymerase chain reaction (RT-PCR). Methods We designed primers specific to the orf1ab and S genes of SARS-CoV-2. Total viral RNA was extracted using the QIAamp Viral RNA Mini Kit. We optimized the RT-LAMP assay. And, this assay was evaluated for its sensitivity and specificity of detection using real-time turbidity monitoring and visual observation. Results The primer sets orf1ab-4 and S-123 amplified the genes in the shortest times, the mean (±SD) time was 18 ± 1.32 min and 20 ± 1.80 min, respectively, and 63°C was the optimum reaction temperature. The sensitivity was 2×101 copies and 2×102 copies per reaction with primer sets orf1ab-4 and S-123, respectively. This assay showed no cross-reactivity with other 60 respiratory pathogens. To describe the availability of this method in clinical diagnosis, we collected 130 specimens from patients with clinically suspected SARS-CoV-2 infection. Among them, 58 were confirmed to be positive and 72 were negative by RT-LAMP. The sensiticity was 100% (95% CI 92.3% - 100%), specificity 100% (95% CI 93.7% - 100%). This assay detected SARS-CoV-2 in the mean (±SD) time of 26.28 ± 4.48 min and the results can be identified with visual observation. Conclusion These results demonstrate that we developed a rapid, simple, specific, and sensitive RT-LAMP assay for SARS-CoV-2 detection among clinical samples. It will be a powerful tool for SARS-CoV-2 identification, and for monitoring suspected patients, close contacts, and high-risk groups.


title: Continuing Medical Student Education During the COVID19 Pandemic: Development of a Virtual Radiation Oncology Clerkship

text: PURPOSE: Our institution cancelled all in-person clerkships due to the COVID-19 pandemic. In response, we designed a virtual radiation oncology medical student clerkship. METHODS AND MATERIALS: We convened an advisory panel to design a virtual clerkship curriculum. We implemented clerkship activities using a cloud-based learning management system, video web conferencing systems and a telemedicine portal. Students completed assessments pre- and post-clerkship to provide data to improve future versions of the clerkship. RESULTS: The virtual clerkship spans 2 weeks and is graded pass or fail. Students attend interactive didactic sessions during the first week and participate in virtual clinic and give talks to the department during the second week. Didactic sessions include lectures, case-based discussions, treatment planning seminars and material adapted from the Radiation Oncology Education Collaborative Study Group curriculum. Students also attend virtual departmental quality assurance rounds, cancer center seminars and multi-disciplinary tumor boards. The enrollment cap was met during the first virtual clerkship period (April 27 through May 8, 2020), with a total of 12 students enrolling. CONCLUSIONS: Our virtual clerkship can increase student exposure and engagement in radiation oncology. Data on clerkship outcomes are forthcoming.


title: Decline in the Use of Surgical Biopsy for Diagnosis of Pulmonary Disease in Hematopoietic Cell Transplantation Recipients in an Era of Improved Diagnostics and Empirical Therapy

text: Abstract Historically, diagnosis of enigmatic pulmonary disease after hematopoietic cell transplantation (HCT) required lung biopsy, but recent advancements in diagnosis and therapy for respiratory infections have changed how clinicians approach pulmonary abnormalities. We examined temporal trends in the use of lung biopsy after HCT. We retrospectively reviewed patients who underwent their first allogeneic HCT at the Fred Hutchinson Cancer Research Center between the years 1993 to 1997, 2003 to 2007, and 2013 to 2015 and subsequently underwent surgical lung biopsy for any reason. Lung biopsy between cohorts were analyzed using a Cox proportional hazards model with death and relapse considered competing risks. Of 1418 patients, 52 (3.7%) underwent 54 post-HCT surgical lung biopsies during 1993 to 1997 compared with 24 (2.1%) and 25 biopsies in the 2003 to 2007 cohort; 2 cases of surgical lung biopsies out of 786 HCT recipients occurred during the 2013 to 2015 cohort (.25%). The median time to biopsy post-HCT was 71.5 days (IQR, 31 to 89) for the early cohort and 97 days (IQR, 42 to 124) for the late cohort, for an overall biopsy incidence of .15 and .075 per 1000 patient days in the first year after HCT, respectively. Patients in the 2003 to 2007 cohort were less likely to undergo a lung biopsy (adjusted HR, .50; 95% CI, .29 to .83; P = .008) when compared with patients in the early cohort, but more patients in the early cohort underwent lung biopsy without antecedent bronchoscopy (25/54 [46%] versus 3/25 [12%], P = .005). Although infections were a more common finding at biopsy in the early cohort (35/1418 versus 8/1148, P < .001), the number of biopsies demonstrating noninfectious lesions was similar between the two cohorts (19/1418 versus 17/1148, P = .76). Fungal infections were the major infectious etiology in both cohorts (32/35 [91%] versus 5/8 [63%], P = .07), but there was a significant reduction in the number of Aspergillus species found at biopsy between the cohorts (30/54 versus 1/25, P < .001). A similar percentage underwent biopsy with therapeutic intent for invasive fungal disease in the 2 cohorts (8/54 [15%] versus 4/25 [16%]). Surgical evaluation of lung disease in HCT recipients significantly declined over a span of 2 decades. The decline from the years 1993 to 1997 compared with 2003 to 2007 was because of a reduction in the number of biopsies for post-transplant infections due to aspergillosis, which is temporally related to improved diagnostic testing by minimally invasive means and the increased use of empiric therapy with extended-spectrum azoles. This practice of primary nonsurgical diagnostic and treatment approaches to pulmonary disease post-HCT have continued, shown by low numbers of surgical biopsies over the last 3 years.


title: Characterization of the duplicate L-SIGN and DC-SIGN genes in miiuy croaker and evolutionary analysis of L-SIGN in fishes

text: Abstract Dendritic cell-specific ICAM-3-grabbing non-integrin (DC-SIGN/CD209) and liver/lymph node-specific ICAM-grabbing non-integrin (L-SIGN/CD299) which are homologues of DC-SIGN are important members in C-type lectin receptors family as key molecules to recognize and eliminate pathogens in the innate immune system. DC-SIGN and L-SIGN have become hot topics in recent studies which both served as cell adhesion and phagocytic pathogen recognition receptors in mammals. However, there have been almost no studies of DC-SIGN and L-SIGN structure and characters in fish, only DC-SIGN in the zebrafish had been studied. In our study, we identified and characterized the full-length miiuy croaker (Miichthys miiuy) DC-SIGN (mmDC-SIGN) and L-SIGN (mmL-SIGN) genes. The sequence analysis results showed that mmDC-SIGN and mmL-SIGN have the same domains with other vertebrates except primates, and share some conserved motifs in CRD among all the vertebrates which play a crucial role in interacting with Ca2+ and for recognizing mannose-containing motifs. Gene synteny of DC-SIGN and L-SIGN were analyzed for the first time and gene synteny of L-SIGN was conserved among the five fishes. Interestingly, one gene next to L-SIGN from gene synteny had high similarity with L-SIGN gene that was described as L-SIGN-like in fish species. While only one L-SIGN gene existed in other vertebrates, two L-SIGN in fish may be in consequence of the fish-specific genome duplication to adapt the specific environment. The evolutionary analysis showed that the ancestral lineages of L-SIGN gene in fishes experienced purifying selection and the current lineages of L-SIGN gene in fishes underwent positive selection, indicating that the ancestral lineages and current lineages of L-SIGN gene in fishes underwent different evolutionary patterns. Both mmDC-SIGN and mmL-SIGN were expressed in all tested tissues and ubiquitously up-regulated in infected liver, spleen and kidney at different sampling time points, indicating that the mmDC-SIGN and mmL-SIGN participated in the immune response to defense against bacteria infection.


title: Navigating Immunosuppression in a Pandemic: A Guide for the Dermatologist from the COVID Task Force of the Medical Dermatology Society and Society of Dermatology Hospitalists

text: Abstract: Dermatologists treating immune-mediated skin disease must now contend with the uncertainties associated with immunosuppressive use in the context of the SARS-CoV-2 pandemic. Though the risk of infection with many commonly used immunosuppressive agents remains low, direct data evaluating the safety of such agents in COVID-19 are scarce. This article reviews and offers guidance based upon currently available safety data, and the most recent COVID-19 outcome data, in patients with immune-mediated dermatologic disease. The interdisciplinary panel of experts emphasize a stepwise, shared decision-making approach in the management of immunosuppressive therapy. The goal of this article is to help providers minimize the risk of disease flares while simultaneously minimizing the risk of iatrogenic harm during an evolving pandemic.


title: New coronavirus outbreak. Lessons learned from the severe acute respiratory syndrome epidemic

text: System dynamics approach offers great potential for addressing how intervention policies can affect the spread of emerging infectious diseases in complex and highly networked systems. Here, we develop a model that explains the severe acute respiratory syndrome coronavirus (SARS-CoV) epidemic that occurred in Hong Kong in 2003. The dynamic model developed with system dynamics methodology included 23 variables (five states, four flows, eight auxiliary variables, six parameters), five differential equations and 12 algebraic equations. The parameters were optimized following an iterative process of simulation to fit the real data from the epidemics. Univariate and multivariate sensitivity analyses were performed to determine the reliability of the model. In addition, we discuss how further testing using this model can inform community interventions to reduce the risk in current and future outbreaks, such as the recently Middle East respiratory syndrome coronavirus (MERS-CoV) epidemic.


title: Recomendaciones para la atención oftalmológica durante el estado de alarma por la pandemia de enfermedad por coronavirus COVID-19

text: ABSTRACT Objective: Minimize exposure to the SARS-CoV-2, reduce the chances of cross-transmission between patients and healthcare personnel, and prevent the development of postoperative complications from the management of patients with eye diseases during the 2019 coronavirus disease pandemic (COVID -19). Methods: COVID-19 literature review and consensus establishment between different Spanish ophthalmology societies in order to provide guidelines and recommendations of maximum resources primarily conditioned by the state of alert, confinement and social distancing that occurs in Spain since March 16, 2020. Results: The recommendations will promote the adoption of action and protection measures for eye care in outpatient clinics, surgical areas and hospitalization, for unconfirmed (asymptomatic and symptomatic) and confirmed COVID-19 patients. Measures must be adapted to the circumstances and availability of Personal Protective Equipment (PPE) in each of the centers and Autonomous Communities, which will be updated according to the pandemic phases and the measures adopted by the Spanish Government. Conclusions: During the COVID-19 pandemic, attention to the potential health risks to the population caused by coronavirus should prevail over the possible progression of the common eye diseases. Ophthalmologists and other eye care professionals must assume a possible progression of these diseases due to the impossibility of adequate patient follow-up.


title: Perforated acute abdomen in a patient with COVID-19: an atypical manifestation of the disease†

text: Abstract Introduction The Coronavirus belongs to a family of RNA viruses that can cause respiratory infection, with the possibility of gastrointestinal manifestations in approximately 5% to 50% of the cases. Objective To report a surgical case with a diagnosis of COVID-19 that developed acute perforated abdomen and pneumothorax. Case report This was an 80-year-old female patient with respiratory symptoms, with dry cough and fever and diffuse abdominal pain with signs of peritonitis. She had leukocytosis, kidney dysfunction and an increase in D-dimer with positive PCR for COVID. Computed tomography of the chest and abdomen showed pneumothorax on the right and extensive pneumoperitoneum Conclusion The presentation of COVID-19 with severe pulmonary and abdominal complications requires specialized and emergency treatments, but it has high mortality rates.


title: Effects of lipopolysaccharide structure on lycopene production in Escherichia coli

text: Abstract Lipopolysaccharides, the major molecules in the outer membrane of Escherichia coli, affect the behavior of bacteria including outer membrane permeability, but its influence on lycopene production in E. coli has never been reported. In this study, the effects of lipopolysaccharides with different structures on lycopene biosynthesis were investigated. Firstly, the heterogenous crtEBI operon were overexpressed in 10 LPS mutant strains of E. coli W3110 (ΔwaaC, ΔwaaF, ΔwaaY, ΔwaaG, ΔwaaR, ΔwaaO, ΔwaaU, ΔwaaP, ΔwaaY and ΔwaaB), and their ability to produce lycopene were compared. ΔwaaC/pWSK29-crtEBI, ΔwaaF/pWSK29-crtEBI and ΔwaaY/pWSK29-crtEBI produced 4.19, 4.20, and 3.81 mg/g lycopene, respectively, while the control W3110/pWSK29-crtEBI produced 3.71 mg/g lycopene; the other strains produced less lycopene than the control. In order to enhance lycopene production, genes dxr, dxr, ispA, and idi were overexpressed in ΔwaaC/pWSK29-crtEBI, ΔwaaF/pWSK29-crtEBI individually or in combination, and the lycopene production in each strain was analyzed. The maximum yield of 5.39 mg/g was achieved in ΔwaaC/pWSK29-crtEBI-SRA, which is 142% higher than that in W3110/pWSK29-crtEBI. The results indicate that the length of lipopolysaccharide affects lycopene biosynthesis in E. coli, and the shorter lipopolysaccharide and higher outer membrane permeability might be beneficial to lycopene biosynthesis.


title: Key Strategies for Clinical Management and Improvement of Healthcare Services for Cardiovascular Disease and Diabetes Patients in the Coronavirus (COVID-19) Settings: Recommendations From the REPROGRAM Consortium

text: Patients with cardiovascular disease and diabetes are at potentially higher risk of infection and fatality due to COVID-19. Given the social and economic costs associated with disability due to these conditions, it is imperative that specific considerations for clinical management of these patients be observed. Moreover, the reorganization of health services around the pandemic response further exacerbates the growing crisis around limited access, treatment compliance, acute medical needs, and mental health of patients in this specific subgroup. Existing recommendations and guidelines emanating from respective bodies have addressed some of the pressure points; however, there are variations and limitations vis a vis patient with multiple comorbidities such as obesity. This article will pull together a comprehensive assessment of the association of cardiovascular disease, diabetes, obesity and COVID-19, its impact on the health systems and how best health systems can respond to mitigate current challenges and future needs. We anticipate that in the context of this pandemic, the cardiovascular disease and diabetes patients need a targeted strategy to ensure the harm to this group does not translate to huge costs to society and to the economy. Finally, we propose a triage and management protocol for patients with cardiovascular disease and diabetes in the COVID-19 settings to minimize harm to patients, health systems and healthcare workers alike.


title: Are chilblain‐like acral skin lesions really indicative of COVID‐19? A prospective study and literature review

text: Recently, young COVID‐19 patients have presented with erythematous and purpuric acral lesions similar to chilblains(1–3). Despite the scarcity of published cases, this topic has attracted significant mass media attention.(4) Dermatologists have noted that more people than would be expected at this time of year are seeking medical attention for these chilblain‐like lesions. Some have suggested that people presenting with this manifestation should be tested and isolated(2). Determining the accuracy of this association is therefore crucial. To establish the real prevalence of COVID‐19 in patients with acral skin lesions, we firstly evaluated all the cases of acral lesions presented in dermatology and paediatrics departments and family doctors’ offices in an eastern Spanish region over a three‐week period. Then we prospectively performed a SARS‐CoV‐2 PCR on nasopharyngeal aspirates taken from all available patients to determine whether their cutaneous manifestations were predictive of a positive result. To put our findings into context, we reviewed all the articles published before May 2020 concerning COVID‐19 patients with cutaneous lesions. We evaluated 58 patients, whose characteristics are summarized in Table 1. In most cases, lesions were chilblain‐like. Fifteen patients had already been tested, and only one had a positive result: an 85‐year‐old man admitted for severe Covid‐19 pneumonia. He had an ulcer on a toe that was finally determined to be vascular in nature. We performed prospective PCR testing in 24 patients. All test results were negative. In total, then, PCR was negative in 38 patients and positive in a single patient whose lesion was very unlikely to be due to COVID‐19. Our bibliography search returned 97 articles and we found 2 more through cross‐references. Nine of these articles dealt with acral lesions specifically. Their results are summarized in Table 2. Data published in the literature are heterogeneous, as are the methods employed to collect them. The first papers looked at cutaneous manifestations in patients with confirmed SARS‐CoV‐2 infection.(5–8) This approach cannot reveal whether this dermatologic manifestation is a specific marker of SARS‐CoV‐2 infection, since patients without COVID‐19 are not included. The other approach, which we have followed, is to analyse all patients with acral lesions. This can be done retrospectively, reporting on patients in the sample who have already been tested, or prospectively, performing the test on all available patients, regardless of whether they have symptoms. The retrospective method has a significant risk of confounding bias: due to the scarcity of COVID‐19 tests(8), they are usually reserved for patients with COVID‐19‐related symptoms, who are obviously more likely to test positive. All previous studies including only patients with cutaneous acral lesions (summarized in table 2) have been retrospective, and only a minority of patients were tested. In total, 12 out of at least 49 tested patients were positive (24.5%). Combined with our results, they total 13 positives out of 88 tests (14.8%). There are two possible explanations for the high proportion of negative results: I) A high number of false negatives. II) The lesions are not related to SARS‐CoV‐2 infection. The low prevalence of an infected contact in our sample, after three weeks of strict confinement in Spain, makes the possibility of being infected in our cohort less likely. The diffusion of this entity by the mass media may have caused patients who would not normally consult to do so.(9) . Other possible explanations include a concomitant parvovirus B19 outbreak(10) or trauma‐induced lesions. Our study suggests that acral skin lesions are not a specific marker of SARS‐CoV‐2 infection. Although larger prospective studies are needed, current evidence indicates that acral skin lesions should not be regarded as a sign of COVID‐19 in otherwise asymptomatic patients.


title: Prevalence of antibodies to specific infectious agents in bovine fetuses from a slaughterhouse in Minnesota

text: Abstract Sera from 486 bovine fetuses, approximately 60 to 270 days of gestation, were collected at slaughter and tested for the presence of immunoglobulins (Ig). One hundred ten (27%) of the sera were positive for IgG and/or IgM. The earliest age at which fetuses tested positive for IgM and IgG was estimated to be 100 and 120 days, respectively. Ig concentration increased with increased age of the fetus. Sera that were positive for Ig were tested for the presence of specific antibodies to five different infectious agents. Bovine parvovirus antibodies were found in 99 of 110 sera (90%) by hemagglutination inhibition (HI) test. However, only 35 (31.8%) of these sera were positive by serum neutralization (SN) test. Antibodies to parainfluenza-3 virus were detected in 30 sera (27%) by HI test and in 20 sera (18%) by SN test. Five (4%) sera contained SN antibodies to bovine viral diarrhea virus. Only one (0.9%) serum sample contained SN antibodies to infectious bovine rhinotracheitis virus. None of the sera had antibodies against five Leptospira spp. Results of this study suggest that bovine parvovirus may be a potential cause of reproductive problems in cattle.


title: Vaccines and Vaccination Practices: Key to Sustainable Animal Production

text: Abstract Protection of food animals from diseases is essential to meet the current and future demands of animal protein for human consumption. Vaccination is a crucial component of the overall strategy to achieve this goal. This article addresses the types of vaccines and vaccination techniques for cattle, swine, poultry, and fish, including brief introductions defining pathogens, immune responses, genetic resistance, and biosecurity.


title: Regulation of Stress Granules and P-Bodies During RNA Virus Infection

text: RNA granules are structures within cells that play major roles in gene expression and homeostasis. Two principle kinds of RNA granules are conserved from yeast to mammals: stress granules (SGs), which contain stalled translation initiation complexes, and processing bodies (P-bodies, PBs), which are enriched with factors involved in RNA turnover. Since RNA granules are associated with silenced transcripts, viruses subvert RNA granule function for replicative advantages. This review, focusing on RNA viruses, discusses mechanisms that manipulate stress granules and P-bodies to promote synthesis of viral proteins. Three main themes have emerged for how viruses manipulate RNA granules; i) cleavage of key host factors, ii) control of PKR activation and iii) redirecting RNA granule components for new or parallel roles in viral reproduction, at the same time disrupting RNA granules. Viruses utilize one or more of these routes to achieve robust and productive infection.


title: Computational simulation of interactions between SARS coronavirus spike mutants and host species-specific receptors

text: Abstract As a critical adaptive mechanism, amino acid replacements on the severe acute respiratory syndrome coronavirus (SARS-CoV) spike protein could alter the receptor-binding specificity of this envelope glycoprotein and in turn lead to the emergence or reemergence of this viral zoonosis. Based on the X-ray structures of SARS-CoV spike receptor-binding domain (RBD) in complex with its functional receptor (angiotensin-converting enzyme 2, ACE2), we perform computational simulations of interactions between three representative RBD mutants and four host species-specific receptors. The comparisons between computational predictions and experimental evidences validate our structural bioinformatics approaches. And the predictions further indicate that some viral prototypes might utilize the rat ACE2 while rats might serve as a vector or reservoir of SARS-CoV.


title: Hygienic hand antiseptics: Should they not have activity and label claims against viruses?

text: Abstract Enteric and respiratory viruses are among the most frequent causes of human infections, and hands play an important role in the spread of these and many other viral diseases. Regular and proper hand hygiene by caregivers and food handlers in particular is essential to decontaminate hands and potentially interrupt such spread. What would be considered a proper decontamination of hands? Handwashing with regular soap and water is often considered sufficient, but what of hygienic handwash and handrub antiseptic products? Are they more effective? The evidence suggests that some clearly are. Activity against bacteria may not reflect the ability of hygienic hand antiseptics to deal with viruses, especially those that are nonenveloped. In spite of the acknowledged importance of hands as vehicles for viruses, there is a lack of suitable regulatory mechanism for handwash or handrub products to make claims of efficacy against viruses. This is in contrast with the ability of general-purpose disinfectants to make antiviral claims, although transmission of viruses from surfaces other than those of reusable medical devices may play only a minor role in virus transmission. This review discusses the (1) recent information on the relative importance of viruses as human pathogens, particularly those causing enteric and respiratory infections; (2) the survival of relevant viruses on human hands in comparison with common gram-negative and gram-positive bacteria; (3) the potential of hands to transfer or receive such contamination on casual contact; (4) role of hands in the spread of viruses; (5) the potential of hygienic measures to eliminate viruses from contaminated hands; (6) relative merits of available protocols to assess the activity of hygienic hand antiseptics against viruses; and (7) factors considered crucial in any tests to assess the activity of hygienic hand antiseptics against viruses. In addition, this review proposes surrogate viruses in such testing and discusses issues for additional consideration by researchers, manufacturers, end-users, and regulators. (Am J Infect Control 2002;30:355-72)


title: A Cross-Sectional Examination on the Factors Related to Emergency Nurses’ Motivation to Protect Themselves against an Ebola Infection

text: Abstract Introduction The 2014-2016 West African Ebola outbreak impacted the United States. Due to the sporadic occurrence of the Ebola infection, there is insufficient research regarding how the United States emergency nurses provide care to patients potentially infected with the Ebola virus and the nurses’ motivation to protect themselves when providing care to these patients. This study was aimed to investigate the predictors of emergency room nurse’s protection motivation. Method A cross-sectional design was employed in the study. A survey developed based on a modified Protection Motivation Theory was administered to randomly selected emergency room nurses who are members of the Emergency Nurses Association. Descriptive statistics, nonparametric Kruskal-Wallis H-Test (as well as post hoc Dunn-Bonferroni test), Spearman Rho correlation, and Stepwise Multiple Linear regression were conducted for data analysis. Result Protection motivation was found in two components: proactive and passive protection motivation. The regression analysis indicated that response efficacy (β=.27, p<.001) and self-efficacy (β=.17, p<.01) significantly predict emergency nurses’ proactive protection motivation, while perceived vulnerability (β=.26, p<.001), response cost (β=.19, p=.001) and knowledge (β=-.15, p<.01) significantly predict emergency nurses’ passive protection motivation. Conclusion Results indicate the need for interventions to improve emergency nurses’ response efficacy, self-efficacy, and knowledge, while simultaneously reducing the nurses’ perceived vulnerability and response cost. Such interventions would be expected to proactively motivate nurses to protect themselves when providing care to patients that exhibit the signs and symptoms of an Ebola infection and reduce their passive protection motivation.


title: Community pharmacists and communication in the time of COVID-19: Applying the health belief model

text: Abstract The emergence of the novel coronavirus disease (COVID-19) pandemic presents an unprecedented health communications challenge. Healthcare providers should reinforce behaviors that limit the spread of the pandemic, including social distancing and remaining in the home whenever possible. Formal communications toolkits may not be prepared in a timely fashion. Community pharmacists can reinforce mitigation behaviors by applying the health belief model (HBM). This commentary provides an overview of the HBM and offers suggestions on how community pharmacists can use it as a guide to patient communication in these uncertain contexts.


title: The emergence of a novel coronavirus (SARS‐CoV‐2) disease and their neuroinvasive propensity may affect in COVID‐19 patients

text: An outbreak of a novel coronavirus (SARS‐CoV‐2) infection has recently emerged and rapidly spreading in humans causing a significant threat to international health and the economy. Rapid assessment and warning are crucial for an outbreak analysis in response to serious public health. SARS‐CoV‐2 shares highly homological sequences with SARS‐CoVs causing highly lethal pneumonia with respiratory distress and clinical symptoms similar to those reported for SARS‐CoV and MERS‐CoV infections. Notably, some COVID‐19 patients also expressed neurologic signs like nausea, headache, and vomiting. Several studies have reported that coronaviruses are not only causing respiratory illness but also invade the central nervous system through a synapse‐connected route. SARS‐CoV infections are reported in both patients and experimental animals' brains. Interestingly, some COVID‐19 patients have shown the presence of SARS‐CoV‐2 virus in their cerebrospinal fluid. Considering the similarities between SARS‐CoV and SARS‐CoV‐2 in various aspects, it remains to clarify whether the potent invasion of SARS‐CoV‐2 may affect in COVID‐19 patients. All these indicate that more detailed criteria are needed for the treatment and the prevention of SARS‐CoV‐2 infected patients. In the absence of potential interventions for COVID‐19, there is an urgent need for an alternative strategy to control the spread of this disease.


title: Letter regarding the article “Enhancing immunity in viral infections, with special emphasis on COVID-19: A review”

text: This letter is aimed to complete the discussion about the potential role of probiotics in COVID-19 patients, also stressing the importance of gut microbiota and the role of the so called gut-lungs axis.


Convalescent plasma – this is no time for competition


title: Inflammation: A bridge between Diabetes and COVID-19, and possible management with sitagliptin

text: Patients with SARS-CoV-2 infections experience lymphopenia and inflammatory cytokine storms in the severe stage of the disease, leading to multi-organ damage. The exact pattern of immune system changes and their condition during the disease process is unclear. The available knowledge has indicated that the NF-kappa-B pathway, which is induced by several mediators, has a significant role in cytokine storm through the various mechanisms. Therefore, identifying the state of the immune cells and the dominant mechanisms for the production of cytokines incorporated in the cytokine storm can be a critical step in the therapeutic approach. On the other hand, some studies identified a higher risk for diabetic patients. Diabetes mellitus exhibits a close association with inflammation and increases the chance of developing COVID-19. Patients with diabetes mellitus have shown to have more virus entry, impaired immunity response, less viral elimination, and dysregulated inflammatory cytokines. The parallel analysis of COVID-19 and diabetes mellitus pathogenesis has proposed that the control of the inflammation through the interfering with the critical points of major signaling pathways may provide the new therapeutic approaches. In recent years, the role of Dipeptidyl Peptidase 4 (DPP4) in chronic inflammation has been proved. Numerous immune cells express the DPP4 protein. DPP4 regulates antibody production, cytokine secretion, and immunoglobulin class switching. DPP4 inhibitors like sitagliptin reduce inflammation intensity in different states. Following the accumulating data, we hypothesize that sitagliptin might reduce COVID-19 severity. Sitagliptin, an available DPP4 inhibitor drug, showed multidimensional anti-inflammatory effects among diabetic patients. It reduces the inflammation mostly by affecting on NF-kappa-B signaling pathway. Under the fact that inflammatory mediators are active in individuals with COVID-19, blocking the predominant pathway could be helpful.


title: SARS‐CoV‐2 lgM/lgG antibody detection confirms the infection after three negative nucleic acid detection

text: An ongoing outbreak of viral pneumonia was caused by a novel coronavirus in China in 2019. By March 19, over 200 thousand confirmed cases of SARS‐CoV‐2 infection and over 9000 deaths have been reported throughout the world. For this infectious disease, nucleic acid detection is still the gold standard for pathogenic detection. However, nucleic acid detection takes a long time and has relatively high "false negative"; therefore, we need urgently a convenient and accurate detection method to make up for this deficiency. In this article, we will show such technical characteristics of lgM/lgG serum antibody detection, compared with nucleic acid detection.


title: Acral cutaneous lesions in the time of COVID‐19

text: Coronavirus disease 2019 (COVID-19) has become a pandemic condition, yet little is known about its dermatologic manifestations. We report here on peculiar (perniosis-like) skin lesions, unreported in the previous years, observed in young outpatients visited in our Dermatologic Unit in the last 4 weeks of COVID-19 pandemic (March-April 2020). Similar cases were referred to us in the same period by Pediatricians and Dermatologists from Italy and European countries.


title: Politicizing the Pandemic: A Schemata Analysis of COVID-19 News in Two Selected Newspapers

text: This article critically studies coronavirus (COVID-19) pandemic news in the press. The article attempts to study the way the news of COVID-19 is used for political and ideological purposes. In order to achieve the aim, two newspapers namely, The New York Times from the United States of America and Global Times from China are selected. Van Dijk’s news schemata framework is used for the analysis of the reports selected from the two newspapers. Van Dijk’s news schemata is crucial for the analysis of any news story (report) whether such a news story is taken from a news channel or a newspaper and whether broadcasted or printed. Based on data analysis, the article found out that the news of COVID-19 has been politicized and used for ideological interests. The article recommends that pandemics should not be politicized, instead we should work together to save our lives and live peacefully.


title: Clinical presentations and outcome of severe acute respiratory syndrome in children

text: Summary Hong Kong has been severely affected by severe acute respiratory syndrome (SARS). Contact in households and healthcare settings is thought to be important for transmission, putting children at particular risk. Most data so far, however, have been for adults. We prospectively followed up the first ten children with SARS managed during the early phase of the epidemic in Hong Kong. All the children had been in close contact with infected adults. Persistent fever, cough, progressive radiographic changes of chest and lymphopenia were noted in all patients. The children were treated with high-dose ribavirin, oral prednisolone, or intravenous methylprednisolone, with no short-term adverse effects. Four teenagers required oxygen therapy and two needed assisted ventilation. None of the younger children required oxygen supplementation. Compared with adults and teenagers, SARS seems to have a less aggressive clinical course in younger children.


title: Inhibitory effects of glycopyrronium, formoterol, and budesonide on coronavirus HCoV-229E replication and cytokine production by primary cultures of human nasal and tracheal epithelial cells

text: Abstract Background Coronavirus 229E (HCoV-229E), one of the causes of the common cold, exacerbates chronic obstructive pulmonary disease (COPD) and bronchial asthma. Long-acting muscarinic antagonists and β2-agonists and inhaled corticosteroids inhibit the exacerbation of COPD and bronchial asthma caused by infection with viruses, including HCoV-229E. However, the effects of these drugs on HCoV-229E replication and infection-induced inflammation in the human airway are unknown. Methods Primary human nasal (HNE) and tracheal (HTE) epithelial cell cultures were infected with HCoV-229E. Results Pretreatment of HNE and HTE cells with glycopyrronium or formoterol decreased viral RNA levels and/or titers, the expression of the HCoV-229E receptor CD13, the number and fluorescence intensity of acidic endosomes where HCoV-229E RNA enters the cytoplasm, and the infection-induced production of cytokines, including IL-6, IL-8, and IFN-β. Treatment of the cells with the CD13 inhibitor 2′2′-dipyridyl decreased viral titers. Pretreatment of the cells with a combination of three drugs (glycopyrronium, formoterol, and budesonide) exerted additive inhibitory effects on viral titers and cytokine production. Pretreatment of HNE cells with glycopyrronium or formoterol reduced the susceptibility to infection, and pretreatment with the three drugs inhibited activation of nuclear factor-kappa B p50 and p65 proteins. Pretreatment with formoterol increased cAMP levels and treatment with cAMP decreased viral titers, CD13 expression, and the fluorescence intensity of acidic endosomes. Conclusions These findings suggest that glycopyrronium, formoterol, and a combination of glycopyrronium, formoterol, and budesonide inhibit HCoV-229E replication partly by inhibiting receptor expression and/or endosomal function and that these drugs modulate infection-induced inflammation in the airway.


title: Incidental COVID-19 in the radiology department: radiographic findings of COVID-19 in asymptomatic patient undergoing CT staging for breast cancer

text: A 59-year old incarcerated woman who was diagnosed with invasive ductal carcinoma in 2016 was brought in for evaluation of the breast cancer. Upon evaluation of the CT chest for breast cancer restaging, diffuse bilateral ground glass opacities and a reverse halo sign in the right lower lobe concerning for atypical viral pneumonia were discovered. The patient was afebrile, had an oxygen saturation of 100%, and denied chest pain as well as shortness of breath. On physical exam, she exhibited decreased breath sounds bilaterally and expiratory wheezing. She later received a COVID-19 test, which came back positive. Infection with the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, also known as COVID-19) may remain asymptomatic in the initial phase, leading to under-recognition and incidental detection on procedures for standard clinical indications. Hospitals, in particular nuclear medicine services, should prepare accordingly in regard to health precautions while keeping in mind the potential discrepancies between clinical presentation and resultant radiologic patterns. This awareness should be heightened in patients at higher risk (i.e. prisoners). Furthermore, by acting upon the incidental detection of this virus during its early stages, subsequent steps could help prevent the spread of the virus.


title: Disinfection of N95 masks artificially contaminated with SARS-CoV-2 and ESKAPE bacteria using hydrogen peroxide plasma: impact on the reutilization of disposable devices

text: INTRODUCTION: One of the serious consequences of the SARS-CoV-2 pandemic is the shortage of protective equipment for health personnel. N95 masks are considered one of the essential protective equipment in the management of patients with COVID-19. The shortage of N95 masks implies potential health risks for health personnel and significant economic losses for the health institution. The objective of this work was to investigate the disinfection of N95 masks artificially contaminated with SARS-CoV-2 and ESKAPE bacteria by using hydrogen peroxide plasma. MATERIAL AND METHODS: We examined the disinfection capacity of hydrogen peroxide plasma against the SARS-CoV-2 and two members of the ESKAPE bacteria (Acinetobacter baumannii and Staphylococcus aureus) through a study of artificial contamination in situ of N95 masks. Amplification of specific genes by RT-PCR of SARS-CoV-2 and microbiological culture of ESKAPE bacteria was performed before and after the disinfection process. RESULTS: SARS-CoV-2 was not detected in all assays using five different concentrations of the virus, and A. baumannii and S. aureus were not cultivable with inoculums of 10(2) to 10(6) CFU after disinfection tests of N95 masks with hydrogen peroxide plasma. CONCLUSION: Disinfection of N95 masks by using the hydrogen peroxide plasma technology can be an alternative for their reuse in a shortage situation. Implications for the use of disinfection technologies of N95 masks and the safety of health personnel are discussed.


title: Exceptional diversity and selection pressure on SARS-CoV and SARS-CoV-2 host receptor in bats compared to other mammals

text: Pandemics originating from pathogen transmission between animals and humans highlight the broader need to understand how natural hosts have evolved in response to emerging human pathogens and which groups may be susceptible to infection. Here, we investigate angiotensin-converting enzyme 2 (ACE2), the host protein bound by SARS-CoV and SARS-CoV-2. We find that the ACE2 gene is under strong selection pressure in bats, the group in which the progenitors of SARS-CoV and SARS-CoV-2 are hypothesized to have evolved, particularly in residues that contact SARS-CoV and SARS-CoV-2. We detect positive selection in non-bat mammals in ACE2 but in a smaller proportion of branches than in bats, without enrichment of selection in residues that contact SARS-CoV or SARS-CoV-2. Additionally, we evaluate similarity between humans and other species in residues that contact SARS-CoV or SARS-CoV-2, revealing potential susceptible species but also highlighting the difficulties of predicting spillover events. This work increases our understanding of the relationship between mammals, particularly bats, and coronaviruses, and provides data that can be used in functional studies of how host proteins are bound by SARS-CoV and SARS-CoV-2 strains.


title: A novel biotinylated nanobody-based blocking ELISA for the rapid and sensitive clinical detection of porcine epidemic diarrhea virus

text: BACKGROUND: Porcine epidemic diarrhea virus (PEDV), which is characterized by severe watery diarrhea, vomiting, dehydration and a high mortality rate in piglets, leads to enormous economic losses to the pork industry and remains a large challenge worldwide. Thus, a rapid and reliable method is required for epidemiological investigations and to evaluate the effect of immunization. However, the current diagnostic methods for PEDV are time-consuming and very expensive and rarely meet the requirements for clinical application. Nanobodies have been used in the clinic to overcome these problems because of the advantages of their easy expression and high level of stability. In the present work, a novel biotinylated nanobody-based blocking ELISA (bELISA) was developed to detect anti-PEDV antibodies in clinical pig serum. RESULTS: Using phage display technology and periplasmic extraction ELISA (PE-ELISA), anti-PEDV N protein nanobodies from three strains of PEDV were successfully isolated after three consecutive rounds of bio-panning from a high quality phage display VHH library. Then, purified Nb2-Avi-tag fusion protein was biotinylated in vitro. A novel bELISA was subsequently developed for the first time with biotinylated Nb2. The cutoff value for bELISA was 29.27%. One hundred and fifty clinical serum samples were tested by both newly developed bELISA and commercial kits. The sensitivity and specificity of bELISA were 100% and 93.18%, respectively, and the coincidence rate between the two methods was 94%. CONCLUSIONS: In brief, bELISA is a rapid, low-cost, reliable and useful nanobody-based tool for the serological evaluation of current PEDV vaccines efficacy and indirect diagnosis of PEDV infection.


THE ETHIC OF CARE, DISABILITY AND REHABILITATION DURING THE COVID-19 PANDEMIC


title: Live poultry market workers are susceptible to both avian and swine influenza viruses, Guangdong Province, China

text: Abstract Guangdong Province is recognized for dense populations of humans, pigs, poultry and pets. In order to evaluate the threat of viral infection faced by those working with animals, a cross-sectional, sero-epidemiological study was conducted in Guangdong between December 2013 and January 2014. Individuals working with swine, at poultry farms, or live poultry markets (LPM), and veterinarians, and controls not exposed to animals were enrolled in this study and 11 (4 human, 3 swine, 3 avian, and 1 canine) influenza A viruses were used in hemagglutination inhibition (HI) assays (7 strains) and the cross-reactivity test (9 strains) in which 5 strains were used in both tests. Univariate analysis was performed to identify which variables were significantly associated with seropositivity. Odds ratios (OR) revealed that swine workers had a significantly higher risk of elevated antibodies against A/swine/Guangdong/L6/2009(H1N1), a classical swine virus, and A/swine/Guangdong/SS1/2012(H1N1), a Eurasian avian-like swine virus than non-exposed controls. Poultry farm workers were at a higher risk of infection with avian influenza H7N9 and H9N2. LPM workers were at a higher risk of infection with 3 subtypes of avian influenza, H5N1, H7N9, and H9N2. Interestingly, the OR also indicated that LPM workers were at risk of H1N1 swine influenza virus infection, perhaps due to the presence of pigs in the LPM. While partial confounding by cross-reactive antibodies against human viruses or vaccines cannot be ruled out, our data suggests that animal exposed people as are more likely to have antibodies against animal influenza viruses.


title: Vitamin D deficiency and COVID-19 pandemic

text: • Vitamin D deficiency is very frequent. • Controlled trials showed that vitamin D decreases acute respiratory infections. • Despite a lack of direct evidence of an effect of vitamin D on COVID-19 infection. • Daily vitamin D supplementation with moderate doses is safe and cheap. • Even a small decrease in COVID-19 infections would easily justify this intervention.


title: Symptom Cluster of ICU nurses treating COVID-19 pneumonia patients in Wuhan, China

text: Abstract Objective In treating highly infectious COVID-19 pneumonia, ICU nurses face a high risk of developing somatic symptom disorder (SSD). The present study aims to investigate the symptoms and causes of SSD of ICU nurses treating COVID-19 pneumonia. The research results are expected to provide evidence for the establishment of a better management strategy. Methods This study enrolled a total of 140 ICU nurses who were selected by Jiangsu Province Hospital to work in Wuhan (the epicenter of the COVID-19 epidemic in China) on 3rd February 2020. A questionnaire “Somatic Symptom Disorders for ICU Nurses in Wuhan No.1 Hospital” was designed based on the “International Classification of Functioning, Disability and Health” (ICF). Exploratory factor analysis was performed to cluster the symptoms, and logistic regression analysis to find the risk factors of the symptoms. Results Five major symptoms were chest-discomfort-and-palpitation (31.4%), dyspnea (30.7%), nausea (21.4%), headache (19.3%), and dizziness (17.9%). In exploratory factor analysis, the symptoms were classified into three clusters: Cluster A of breathing and sleep disturbances (dizziness, sleepiness, dyspnea); Cluster B of gastrointestinal complaints and pain (nausea, headache), and Cluster C of general symptoms (xerostomia, fatigue, chest-discomfort-and-palpitation). In Cluster A, urine/feces splash, sex, and sputum splash were independent predictive factors. In Cluster B, fall of protective glasses and urine/feces splash were independent predictive factors. In Cluster C, urine/feces splash and urine/feces clearance were independent predictive factors. Conclusion The ICU nurses in Wuhan showed varying and overlapping SSDs. These SSDs could be classified into three symptom clusters. Based on the characteristics of their SSDs, specific interventions could be implemented to safeguard the health of ICU nurses.


title: An Enzymatic TMPRSS2 Assay for Assessment of Clinical Candidates and Discovery of Inhibitors as Potential Treatment of COVID-19

text: SARS-CoV-2 is the viral pathogen causing the COVID19 global pandemic. Consequently, much research has gone into the development of pre-clinical assays for the discovery of new or repurposing of FDA-approved therapies. Preventing viral entry into a host cell would be an effective antiviral strategy. One mechanism for SARS-CoV-2 entry occurs when the spike protein on the surface of SARS-CoV-2 binds to an ACE2 receptor followed by cleavage at two cut sites (“priming”) that causes a conformational change allowing for viral and host membrane fusion. This fusion event is proceeded by release of viral RNA within the host cell. TMPRSS2 has an extracellular protease domain capable of cleaving the spike protein to initiate membrane fusion. Additionally, knock-out studies in mice have demonstrated reduced infection in the absence of TMPRSS2 with no detectable physiological impact; thus, TMPRSS2 is an attractive target for therapeutic development. A validated inhibitor of TMPRSS2 protease activity would be a valuable tool for studying the impact TMPRSS2 has in viral entry and potentially be an effective antiviral therapeutic. To enable inhibitor discovery and profiling of FDA-approved therapeutics, we describe an assay for the biochemical screening of recombinant TMPRSS2 suitable for high throughput application. We demonstrate effectiveness to quantify inhibition down to subnanomolar concentrations by assessing the inhibition of camostat, nafamostat and gabexate, clinically approved agents in Japan for pancreatitis due to their inhibition of trypsin-like proteases. Nafamostat and camostat are currently in clinical trials against COVID19. The rank order potency for the three inhibitors is: nafamostat (IC(50) = 0.27 nM), camostat (IC(50) = 6.2 nM) and gabexate (IC(50) = 130 nM). Further profiling of these three inhibitors against a panel of proteases provides insight into selectivity and potency.


title: Prediction for the spread of COVID-19 in India and effectiveness of preventive measures

text: Abstract The spread of COVID-19 in the whole world has put the humanity at risk. The resources of some of the largest economies are stressed out due to the large infectivity and transmissibility of this disease. Due to the growing magnitude of number of cases and its subsequent stress on the administration and health professionals, some prediction methods would be required to predict the number of cases in future. In this paper, we have used data-driven estimation methods like long short-term memory (LSTM) and curve fitting for prediction of the number of COVID-19 cases in India 30 days ahead and effect of preventive measures like social isolation and lockdown on the spread of COVID-19. The prediction of various parameters (number of positive cases, number of recovered cases, etc.) obtained by the proposed method is accurate within a certain range and will be a beneficial tool for administrators and health officials.


title: Data Quality of Chinese Surveillance of COVID-19: Objective Analysis Based on WHO’s Situation Reports

text: Was there quality in the Chinese epidemiological surveillance system during the COVID-19 pandemic? Using data of World Health Organization’s situation reports (until situation report 55), an objective analysis was realized to answer this important question. Fulfillment of Benford’s law (first digit law) is a rapid tool to suggest good data quality. Results suggest that China had an acceptable quality in its epidemiological surveillance system. Furthermore, more detailed and complete analyses could complement the evaluation of the Chinese surveillance system.


Kann und darf das Handy gegen Corona helfen? : Pressemitteilung des Landesbeauftragten für den Datenschutz und die Informationsfreiheit Rheinland-Pfalz vom 31.03.2020


title: Risk factors for acute respiratory tract infections in general practitioner patients in The Netherlands: a case-control study

text: BACKGROUND: Acute respiratory tract infections (ARTI) are an important public health problem. Improved identification of risk factors might enable targeted intervention. Therefore we carried out a case-control study with the aim of identifying environmental risk factors for ARTI consultations in the Dutch general population. METHODS: A subset of patients visiting their GP in the period of 2000–2003 with an ARTI (cases) and age-matched controls (visiting for other complaints) were included in a case-control study. They were asked to complete a questionnaire about potential risk factors. Conditional logistic regression was used to calculate odds ratio's (OR) and 95% confidence intervals (CI) to estimate the independent effect of potential risk factors. RESULTS: A total of 493 matched pairs of case and control subjects were enrolled. Exposure to persons with respiratory complaints, both inside and outside the household, was found to be an independent risk factor for visiting a GP with an ARTI (respectively OR(adj )= 1.9 and OR(adj )= 3.7). Participants exposed to dampness or mould at home (OR(adj)=0.5) were significantly less likely to visit their GP. In accordance with the general risk of consultations for ARTI, participants with a laboratory-confirmed ARTI who were exposed to persons with respiratory complaints outside the household were also significantly more likely to visit their GP (OR(adj)=2.5). CONCLUSION: This study confirmed that heterogeneity in the general population as well as in pathogens causing ARTI makes it complicated to detect associations between potential risk factors and respiratory infections. Whereas it may be difficult to intervene on the risk posed by exposure to persons with respiratory complaints, transmission of ARTI in the general population might be reduced by improved hygienic conditions.