title: Pathological study of the 2019 novel coronavirus disease (COVID-19) through postmortem core biopsies

text: Data on pathologic changes of the 2019 novel coronavirus disease (COVID-19) are scarce. To gain knowledge about the pathology that may contribute to disease progression and fatality, we performed postmortem needle core biopsies of lung, liver, and heart in four patients who died of COVID-19 pneumonia. The patients’ ages ranged from 59 to 81, including three males and one female. Each patient had at least one underlying disease, including immunocompromised status (chronic lymphocytic leukemia and renal transplantation) or other conditions (cirrhosis, hypertension, and diabetes). Time from disease onset to death ranged from 15 to 52 days. All patients had elevated white blood cell counts, with significant rise toward the end, and all had lymphocytopenia except for the patient with leukemia. Histologically, the main findings are in the lungs, including injury to the alveolar epithelial cells, hyaline membrane formation, and hyperplasia of type II pneumocytes, all components of diffuse alveolar damage. Consolidation by fibroblastic proliferation with extracellular matrix and fibrin forming clusters in airspaces is evident. In one patient, the consolidation consists of abundant intra-alveolar neutrophilic infiltration, consistent with superimposed bacterial bronchopneumonia. The liver exhibits mild lobular infiltration by small lymphocytes, and centrilobular sinusoidal dilation. Patchy necrosis is also seen. The heart shows only focal mild fibrosis and mild myocardial hypertrophy, changes likely related to the underlying conditions. In conclusion, the postmortem examinations show advanced diffuse alveolar damage, as well as superimposed bacterial pneumonia in some patients. Changes in the liver and heart are likely secondary or related to the underlying diseases.


title: Comovement amongst the demand for New Zealand tourism

text: Tourism research is replete with applications of univariate time-series decomposition techniques: multivariate frameworks have been largely ignored. In this paper, we employ a common-feature-based, multivariate trend-cycle decomposition approach to examine common trends and common cycles amongst the demand for New Zealand tourism originating in Australia, China, the US, the UK, as well as other tourism-importing countries considered as one group: Others. Upon isolating trends and cycles in tourism demand from these countries, we find evidence of strong long-term comovement: they share one common trend. We also find evidence of short-term comovement, albeit to a lesser extent: four common cycles emerge; the cyclical patterns in tourism demand from Australia, China, the US, and Others are strongly correlated.


title: Recombinant SARS-CoV-2 spike S1-Fc fusion protein induced high levels of neutralizing responses in nonhuman primates

text: Abstract The COVID-19 outbreak has become a global pandemic responsible for over 2,000,000 confirmed cases and over 126,000 deaths worldwide. In this study, we examined the immunogenicity of CHO-expressed recombinant SARS-CoV-2 S1-Fc fusion protein in mice, rabbits, and monkeys as a potential candidate for a COVID-19 vaccine. We demonstrate that the S1-Fc fusion protein is extremely immunogenic, as evidenced by strong antibody titers observed by day 7. Strong virus neutralizing activity was observed on day 14 in rabbits immunized with the S1-Fc fusion protein using a pseudovirus neutralization assay. Most importantly, in less than 20 days and three injections of the S1-Fc fusion protein, two monkeys developed higher virus neutralizing titers than a recovered COVID-19 patient in a live SARS-CoV-2 infection assay. Our data strongly suggests that the CHO-expressed SARS-CoV-2 S1-Fc recombinant protein could be a strong candidate for vaccine development against COVID-19.


title: Mucosal immunity: The immunology of breast milk

text: Abstract The mammary glands represent one part of the mucosal immune system, a definable, subunit of humoral and cellular immune functions in man that appears to have developed particular qualities well suited to guard our interface with the environment. As our understanding of secretory immunoglobulins and lymphocyte migration patterns continues to develop, the immunologic components found in breast milk appear increasingly likely to play a specific immunologic role in the protection of the nursing infant. The biologic basis for the observed protective effect of breast-feeding is reviewed with an emphasis on the mechanisms involved in the development and maintenance of mucosal immunity in general.


title: Emerging Prevention and Treatment Strategies to Control COVID-19

text: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), has now become a serious global threat after inflicting more than 8 million infections and 425,000 deaths in less than 6 months. Currently, no definitive treatment or prevention therapy exists for COVID-19. The unprecedented rise of this pandemic has rapidly fueled research efforts to discover and develop new vaccines and treatment strategies against this novel coronavirus. While hundreds of vaccines/therapeutics are still in the preclinical or early stage of clinical development, a few of them have shown promising results in controlling the infection. Here, in this review, we discuss the promising vaccines and treatment options for COVID-19, their challenges, and potential alternative strategies.


title: Cytokine and Chemokine Levels in Patients with Severe Fever with Thrombocytopenia Syndrome Virus

text: BACKGROUND: Severe fever with thrombocytopenia syndrome virus (SFTSV), which can cause hemorrhagic fever–like illness, is a newly discovered bunyavirus in China. The pathogenesis of SFTSV infection is poorly understood. However, it has been suggested that immune mechanisms, including cytokines and chemokines, play an important role in disease pathogenesis. In the present study, we investigated host cytokine and chemokine profiles in serum samples of patients with SFTSV infection from Northeast China and explored a possible correlation between cytokine levels and disease severity. METHODS AND PRINCIPAL FINDINGS: Acute phase serum samples from 40 patients, diagnosed with SFTSV infection were included. Patients were divided into two groups – severe or non-severe – based on disease severity. Levels of tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, interleukin-6, interferon (IFN)-γ, IFN- γ-induced protein (IP)-10 and RANTES were measured in the serum samples with commercial ELISAs. Statistical analysis showed that increases in TNF-α, IP-10 and IFN-γ were associated with disease severity. CONCLUSIONS: We suggest that a cytokine-mediated inflammatory response, characterized by cytokine and chemokine production imbalance, might be in part responsible for the disease progression of patients with SFTSV infection.


title: Going to Bat(s) for Studies of Disease Tolerance

text: A majority of viruses that have caused recent epidemics with high lethality rates in people, are zoonoses originating from wildlife. Among them are filoviruses (e.g., Marburg, Ebola), coronaviruses (e.g., SARS, MERS), henipaviruses (e.g., Hendra, Nipah) which share the common features that they are all RNA viruses, and that a dysregulated immune response is an important contributor to the tissue damage and hence pathogenicity that results from infection in humans. Intriguingly, these viruses also all originate from bat reservoirs. Bats have been shown to have a greater mean viral richness than predicted by their phylogenetic distance from humans, their geographic range, or their presence in urban areas, suggesting other traits must explain why bats harbor a greater number of zoonotic viruses than other mammals. Bats are highly unusual among mammals in other ways as well. Not only are they the only mammals capable of powered flight, they have extraordinarily long life spans, with little detectable increases in mortality or senescence until high ages. Their physiology likely impacted their history of pathogen exposure and necessitated adaptations that may have also affected immune signaling pathways. Do our life history traits make us susceptible to generating damaging immune responses to RNA viruses or does the physiology of bats make them particularly tolerant or resistant? Understanding what immune mechanisms enable bats to coexist with RNA viruses may provide critical fundamental insights into how to achieve greater resilience in humans.


title: Pseudotyped Vesicular Stomatitis Virus for Analysis of Virus Entry Mediated by SARS Coronavirus Spike Proteins

text: Severe acute respiratory syndrome (SARS) coronavirus (CoV) contains a spike (S) protein that binds to a receptor molecule (angiotensin-converting enzyme 2; ACE2), induces membrane fusion, and serves as a neutralizing epitope. To study the functions of the S protein, we describe here the generation of SARS-CoV S protein-bearing vesicular stomatitis virus (VSV) pseudotype using a VSV∆G∗/GFP system in which the G gene is replaced by the green fluorescent protein (GFP) gene (VSV-SARS-CoV-St19/GFP). Partial deletion of the cytoplasmic domain of SARS-CoV S protein (SARS-CoV-St19) allowed efficient incorporation into the VSV particle that enabled the generation of a high titer of pseudotype virus. Neutralization assay with anti-SARS-CoV antibody revealed that VSV-SARS-St19/GFP pseudotype infection is mediated by SARS-CoV S protein. The VSV∆G∗/SEAP system, which secretes alkaline phosphatase instead of GFP, was also generated as a VSV pseudotype having SARS-CoV S protein (VSV-SARS-CoV-St19/SEAP). This system enabled high-throughput analysis of SARS-CoV S protein-mediated cell entry by measuring alkaline phosphatase activity. Thus, VSV pseudotyped with SARS-CoV S protein is useful for developing a rapid detection system for neutralizing antibody specific for SARS-CoV infection as well as studying the S-mediated cell entry of SARS-CoV.


title: Letter: Covid‐19, and vitamin D. Authors' reply

text: LINKED CONTENT This article is linked to Sands et al papers. To view these articles, visit https://doi.org/10.1111/apt.15555 and https://doi.org/10.1111/apt.15716.


title: Coronavirus-induced demyelination occurs in the absence of CD28 costimulatory signals

text: Abstract Infection of mice with mouse hepatitis virus (MHV) strain A59 results in acute encephalitis, hepatitis, and chronic demyelinating disease. T lymphocytes play an important role in MHV infection, and costimulatory signals are an important component of T cell function. To elucidate the role of the main costimulatory molecule, CD28, in MHV pathogenesis and demyelination, we examined the kinetics of MHV-A59 infection in CD28 knockout mice. MHV-A59-infected CD28 knockout mice developed acute encephalitis and hepatitis, and the same degree of chronic demyelination as normal C57Bl/6 (B6) mice. Thus, CD28, the costimulatory T cell molecule, is not required for MHV infection and MHV-induced demyelination.


title: Local Antigen Encounter Is Essential for Establishing Persistent CD8(+) T-Cell Memory in the CNS

text: While the brain is considered an immune-privileged site, the CNS may nevertheless be the focus of immune mediated inflammation in the case of infection and certain autoimmune diseases, e.g., multiple sclerosis. As in other tissues, it has been found that acute T-cell infiltration may be followed by establishment of persistent local T-cell memory. To improve our understanding regarding the regulation of putative tissue resident memory T (Trm) cells in CNS, we devised a new model system for studying the generation of Trm cells in this site. To this purpose, we exploited the fact that the CNS may be a sanctuary for adenoviral infection, and to minimize virus-induced disease, we chose replication-deficient adenoviruses for infection of the CNS. Non-replicating adenoviruses are known to be highly immunogenic, and our studies demonstrate that intracerebral inoculation causes marked local T-cell recruitment, which is followed by persistent infiltration of the CNS parenchyma by antigen specific CD8(+) T cells. Phenotypical analysis of CNS infiltrating antigen specific CD8(+) T cells was consistent with these cells being Trms. Regarding the long-term stability of the infiltrate, resident CD8(+) T cells expressed high levels of the anti-apoptotic molecule Bcl-2 as well as the proliferation marker Ki-67 suggesting that the population is maintained through steady homeostatic proliferation. Functionally, memory CD8(+) T cells from CNS matched peripheral memory cells with regard to capacity for ex vivo cytotoxicity and cytokine production. Most importantly, our experiments revealed a key role for local antigen encounter in the establishment of sustained CD8(+) T-cell memory in the brain. Inflammation in the absence of cognate antigen only led to limited and transient infiltration by antigen specific CD8(+) T cells. Together these results indicate that memory CD8(+) T cells residing in the CNS predominantly mirror previous local infections and immune responses to local autoantigens.


title: INITIAL HIGH VIRAL LOAD IS ASSOCIATED WITH PROLONGED SHEDDING OF HUMAN RHINOVIRUS IN ALLOGENEIC HEMATOPOIETIC CELL TRANSPLANT RECIPIENTS

text: INTRODUCTION: Recent data suggest human rhinovirus (HRV) is associated with lower respiratory tract infection and mortality in hematopoietic cell transplant (HCT) recipients. Examining risk factors for prolonged viral shedding may provide critical insight for the development of novel therapeutics and help inform infection prevention practices. Our objective was to identify risk factors for prolonged shedding of HRV post-HCT. MATERIALS AND METHODS: We prospectively collected weekly nasal samples from allogeneic HCT recipients from day 0 to day 100 post-transplant, and performed realtime RT-PCR (12/2005 to 2/2010). Subjects with symptomatic HRV infection and a negative test within 2 weeks of the last positive were included. Duration of shedding was defined as time between the first positive and first negative samples. Cycle threshold (Ct) values were used as a proxy for viral load. HRV species were identified by sequencing the 5’ non-coding region. Logistic regression analyses were performed to evaluate factors associated with prolonged shedding (≥21 days). RESULTS: We identified 38 HCT recipients with HRV infection fulfilling study criteria (32 adults, 6 children). Median duration of shedding was 9.5 days (2-89 days); 18 patients had prolonged shedding. Among 26 samples sequenced, 69% were species A, and species B and C accounted for 15% each; the median shedding duration of HRV did not differ between species (P = 0.17). Bivariable logistic regression analyses suggest that initial high viral load (low Ct value) is associated with prolonged shedding. CONCLUSIONS: HCT recipients with initial high viral loads are at risk for prolonged HRV viral shedding.


How to Perform Pediatric Lung Ultrasound Examinations in the Time of COVID‐19


title: Challenges amid COVID-19 times - Review of the changing practices in a clinical chemistry laboratory from a developing country

text: Corona Virus Disease 2019 (COVID-19) pandemic is the defining global health crisis of our time. Compared with its neighbors China and Iran, which were rated as epi-centers of the outbreak, Pakistan has lower standards of health care, unstable economy and dearth of financial resources to tackle the outbreak. Like other institutes and industries in the country, clinical laboratories were succumbed to a variety of challenges. This article is based on the experience and adapted workflow measures from the Clinical Chemistry laboratory at the Aga Khan University Hospital (AKUH), Karachi, which serves as a national referral center with its widespread network of satellite laboratories and phlebotomy centers across the country. It highlights the challenges faced and the appropriate responses to ensure the provision of diagnostic facilities during the COVID-19 outbreak. Furthermore, the lessons acquired and necessary preparations for the post crisis situation are also incorporated.


title: Cloning, expression and characterization of ferret CXCL10

text: Chemokines and their receptors function in the recruitment and activation of cells of the immune system to sites of inflammation. As such, chemokines play an important role in mediating pathophysiological events during microbial infection. In particular, CXCL9, 10 and 11 and their cognate receptor CXCR3 have been associated with the clinical course of several infectious diseases, including severe acute respiratory syndrome (SARS) and influenza. While CXCL9, 10 and 11 share the same receptor and have overlapping functions, each can also have unique activity in host defense. The lack of a preferred characterized animal model for SARS has brought our attention to ferrets, which have been used for years in influenza studies. The lack of immunological reagents for ferrets prompted us to clone CXCL9, 10, 11 and CXCR3 and, in the case of CXCL10, to express the gene as a recombinant protein. In this study we demonstrate that endogenous ferret CXCL10 exhibits similar mRNA expression patterns in the lungs of deceased SARS patients and ferrets experimentally infected with SARS coronavirus. This study therefore represents an important step towards development of the ferret as a model for the role of CXCL9, 10 and 11:CXCR3 axis in severe viral infections.


The Novel Coronavirus COVID-19 Outbreak: Global Implications for Antimicrobial Resistance


title: Emergency remote teaching during Coronavirus pandemic: the current trend and future directive at Middle East College Oman

text: Due to COVID-19 outbreaks worldwide, the academic institutions have been enforced to entirely cancel face-to-face teaching including laboratories and other learning experiences as a mitigation step against the risk posed by the Coronavirus. Accordingly, various measures by the higher education providers have been initiated to implement social isolation strategies, and online teaching is followed with rapid curriculum transformation. The online delivery is more convenient, as it can provide vibrant and dynamic teaching and learning environment. However, due to time constraint, the curriculum transformation is anticipated to occur rapidly without sufficient preparation. Therefore, in this study, the concept of the emergency remote teaching (ERT) including its application and evaluation is thoroughly discussed. The application of the ERT in the Middle East College Oman has been considered as case study. This study draws on CIPP evaluation model to assess the effectiveness of the adopted model, and qualitative data were collected online taking random samples of students and educators. Besides, interview and questionnaire responses, experiences, beliefs, and challenges encountered by the educators and students on the emergency remote teaching were used and analyzed. In addition, students’ weekly attendance segregated with session modes, levels and module nature were considered to evaluate the students’ participation to the online classes. The collected information were analyzed, and based on the analysis outcomes, recommendations were forwarded to serve as an input for future strategies and policies and to improve the performance of teaching learning activities during similar circumstances.


Old Master Zhu: in memory of virologist Guan-Fu Zhu


title: Report of EEG Finding on Critically Ill Patients with COVID‐19

text: In March 2020, we treated a cohort of 26 critically ill hospitalized SARS‐CoV‐2 infected patients who received EEGs to assess unexplained altered mental status, loss of consciousness, or poor arousal and responsiveness. Of the 26 patients studied, 5 patients had EEGs that showed Periodic Discharges (PD) consisting of high amplitude frontal monomorphic delta waves with absence of epileptic activity. These findings may suggest CNS injury potentially related to COVID‐19 in these patients. This article is protected by copyright. All rights reserved.


title: Clinical and virological data of the first cases of COVID-19 in Europe: a case series

text: BACKGROUND: On Dec 31, 2019, China reported a cluster of cases of pneumonia in people at Wuhan, Hubei Province. The responsible pathogen is a novel coronavirus, named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We report the relevant features of the first cases in Europe of confirmed infection, named coronavirus disease 2019 (COVID-19), with the first patient diagnosed with the disease on Jan 24, 2020. METHODS: In this case series, we followed five patients admitted to Bichat-Claude Bernard University Hospital (Paris, France) and Pellegrin University Hospital (Bordeaux, France) and diagnosed with COVID-19 by semi-quantitative RT-PCR on nasopharyngeal swabs. We assessed patterns of clinical disease and viral load from different samples (nasopharyngeal and blood, urine, and stool samples), which were obtained once daily for 3 days from hospital admission, and once every 2 or 3 days until patient discharge. All samples were refrigerated and shipped to laboratories in the National Reference Center for Respiratory Viruses (The Institut Pasteur, Paris, and Hospices Civils de Lyon, Lyon, France), where RNA extraction, real-time RT-PCR, and virus isolation and titration procedures were done. FINDINGS: The patients were three men (aged 31 years, 48 years, and 80 years) and two women (aged 30 years and 46 years), all of Chinese origin, who had travelled to France from China around mid-January, 2020. Three different clinical evolutions are described: (1) two paucisymptomatic women diagnosed within a day of exhibiting symptoms, with high nasopharyngeal titres of SARS-CoV-2 within the first 24 h of the illness onset (5·2 and 7·4 log(10) copies per 1000 cells, respectively) and viral RNA detection in stools; (2) a two-step disease progression in two young men, with a secondary worsening around 10 days after disease onset despite a decreasing viral load in nasopharyngeal samples; and (3) an 80-year-old man with a rapid evolution towards multiple organ failure and a persistent high viral load in lower and upper respiratory tract with systemic virus dissemination and virus detection in plasma. The 80-year-old patient died on day 14 of illness (Feb 14, 2020); all other patients had recovered and been discharged by Feb 19, 2020. INTERPRETATION: We illustrated three different clinical and biological types of evolution in five patients infected with SARS-CoV-2 with detailed and comprehensive viral sampling strategy. We believe that these findings will contribute to a better understanding of the natural history of the disease and will contribute to advances in the implementation of more efficient infection control strategies. FUNDING: REACTing (Research & Action Emerging Infectious Diseases).


title: Coverage of Health Information by Different Sources in Communities: Implication for COVID-19 Epidemic Response

text: Health personnel and community workers are at the front line of the COVID-19 emergency response and need to be equipped with adequate knowledge related to epidemics for an effective response. This study aimed to identify the coverage of COVID-19 health information via different sources accessed by health workers and community workers in Vietnam. A cross-sectional study using a web-based survey was carried out from January to February 2020 in Vietnam. Respondent-driven sampling (RDS) was used for recruiting participants. We utilized the exploratory factor analysis (EFA) to examine the construct validity of the questionnaire. A higher percentage of participants knew about “Clinical and pathogen characteristics of COVID-19”, compared to “Regulations and policies related to COVID-19”. The percentage of participants accessing the information on “Guidelines and policies on prevention and control of COVID-19” was the lowest, especially among medical students. “Mass media and peer-educators” channels had a higher score of accessing COVID-19 information, compared to “Organizations/ agencies/ associations” sources. Participants consumed most of their COVID-19 information via “Internet, online newspapers, social networks”. Our findings indicate an urgency to re-design training programs and communication activities for a more effective dissemination of information related to the COVID-19 epidemic or epidemics in general.


title: Encephalitogenicity of myelin basic protein exon-2 peptide in mice

text: Abstract Immunization with a synthetic peptide with an amino acid sequence corresponding to mouse myelin basic protein exon-2 induced mild experimental allergic encephalitis (EAE) in B10.RIII mice, very mild disease in SJL/J mice and no disease in (SJL × PL)F1 hybrid mice. In contrast, adoptive transfer of an exon-2 peptide-specific T cell line from SJL mice induced severe relapsing EAE in syngeneic recipients. The T cell line was specific for exon-2 peptide and did not cross-react appreciably with an MBP preparation consisting of the 18.5 and 14-kDa isoforms. mRNA for exon-2 containing isoforms could be demonstrated in the spinal cord of SJL/J and B10.RIII mice by amplification using exon-2 and exon-4 oligonucleotide primers. On a relative basis, the level of exon-2 cDNA was lower than that of exon-1 cDNA in the same spinal cord preparations from both strains of mice.


title: Structural genomics and interactomics of 2019 Wuhan novel coronavirus, 2019-nCoV, indicate evolutionary conserved functional regions of viral proteins

text: During its first month, the recently emerged 2019 Wuhan novel coronavirus (2019-nCoV) has already infected many thousands of people in mainland China and worldwide and took hundreds of lives. However, the swiftly spreading virus also caused an unprecedentedly rapid response from the research community facing the unknown health challenge of potentially enormous proportions. Unfortunately, the experimental research to understand the molecular mechanisms behind the viral infection and to design a vaccine or antivirals is costly and takes months to develop. To expedite the advancement of our knowledge we leverage the data about the related coronaviruses that is readily available in public databases, and integrate these data into a single computational pipeline. As a result, we provide a comprehensive structural genomics and interactomics road-maps of 2019-nCoV and use these information to infer the possible functional differences and similarities with the related SARS coronavirus. All data are made publicly available to the research community at http://korkinlab.org/wuhan


title: SARS coronavirus entry into host cells through a novel clathrin- and caveolae-independent endocytic pathway

text: While severe acute respiratory syndrome coronavirus (SARS-CoV) was initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that virus entry may also involve endocytosis. We have found that SARS-CoV enters cells via pH- and receptor-dependent endocytosis. Treatment of cells with either SARS-CoV spike protein or spike-bearing pseudoviruses resulted in the translocation of angiotensin-converting enzyme 2 (ACE2), the functional receptor of SARS-CoV, from the cell surface to endosomes. In addition, the spike-bearing pseudoviruses and early endosome antigen 1 were found to colocalize in endosomes. Further analyses using specific endocytic pathway inhibitors and dominant-negative Eps15 as well as caveolin-1 colocalization study suggested that virus entry was mediated by a clathrin- and caveolae-independent mechanism. Moreover, cholesterol- and sphingolipid-rich lipid raft microdomains in the plasma membrane, which have been shown to act as platforms for many physiological signaling pathways, were shown to be involved in virus entry. Endocytic entry of SARS-CoV may expand the cellular range of SARS-CoV infection, and our findings here contribute to the understanding of SARS-CoV pathogenesis, providing new information for anti-viral drug research. SUPPLEMENTARY INFORMATION: The online version of this article (doi:10.1038/cr.2008.15) contains supplementary material, which is available to authorized users.


title: Surgery during COVID-19 crisis conditions: can we protect our ethical integrity against the odds?

text: COVID-19 is reducing the ability to perform surgical procedures worldwide, giving rise to a multitude of ethical, practical and medical dilemmas. Adapting to crisis conditions requires a rethink of traditional best practices in surgical management, delving into an area of unknown risk profiles. Key challenging areas include cancelling elective operations, modifying procedures to adapt local services and updating the consenting process. We aim to provide an ethical rationale to support change in practice and guide future decision-making. Using the four principles approach as a structure, Medline was searched for existing ethical frameworks aimed at resolving conflicting moral duties. Where insufficient data were available, best guidance was sought from educational institutions: National Health Service England and The Royal College of Surgeons. Multiple papers presenting high-quality, reasoned, ethical theory and practice guidance were collected. Using this as a basis to assess current practice, multiple requirements were generated to ensure preservation of ethical integrity when making management decisions. Careful consideration of ethical principles must guide production of local guidance ensuring consistent patient selection thus preserving equality as well as quality of clinical services. A critical issue is balancing the benefit of surgery against the unknown risk of developing COVID-19 and its associated complications. As such, the need for surgery must be sufficiently pressing to proceed with conventional or non-conventional operative management; otherwise, delaying intervention is justified. For delayed operations, it is our duty to quantify the long-term impact on patients’ outcome within the constraints of pandemic management and its long-term outlook.


title: COVID-19 in Italy: Dataset of the Italian Civil Protection Department

text: The database here described contains data of integrated surveillance for the “Coronavirus disease 2019” (abbreviated as COVID-19 by the World Health Organization) in Italy, caused by the novel coronavirus SARS-CoV-2. The database, included in a main folder called COVID-19, has been designed and created by the Italian Civil Protection Department, which currently manages it. The database consists of six folders called ‘aree’ (containing charts of geographical areas interested by containment measures), ‘dati-andamento-nazionale’ (containing data relating to the national trend of SARS-CoV-2 spread), ‘dati-json’ (containing data that summarize the national, provincial and regional trends of SARS-CoV-2 spread), ‘dati-province’ (containing data relating to the provincial trend of SARS-CoV-2 spread), ‘dati-regioni’ (containing data relating to the regional trend of SARS-CoV-2 spread) and ‘schede-riepilogative’ (containing summary sheets relating to the provincial and regional trends of SARS-CoV-2 spread). The Italian Civil Protection Department daily receives data by the Italian Ministry of Health, analyzes them and updates the database. Thus, the database is subject to daily updates and integrations. The database is freely accessible (CC-BY-4.0 license) at https://github.com/pcm-dpc/COVID-19. This database is useful to provide insight on the spread mechanism of SARS-CoV-2, to support organizations in the evaluation of the efficiency of current prevention and control measures, and to support governments in the future prevention decisions.


title: Studies in neurobiology and aging at the United States National Institutes of health‐sponsored regional primate research centers

text: The seven NIH‐sponsored Regional Primate Research Centers conduct a wide variety of studies in the neurosciences and on aging. Monkeys and chimpanzees are investigated to validate the results of research obtained with lower species. In addition, primates are often the only species with the appropriate characteristics for studying a particular health problem or certain behavioral or biological principles. One neurological disorder for which the primate model has proven crucial is Parkinson's disease. The surgical implantation of dopaminergic cells and tissue containing a growth factor has been developed in the primate model, and is now being evaluated clinically in humans with Parkinsonism. In addition to neurological diseases such as Parkinson's, multiple sclerosis, and epilepsy, the visual system is a strong focus of basic and applied research at the Centers. In 1981, two neuroscientists whose basic research on vision with application to pediatric ophthalmology took place in part at the New England Regional Primate Research Center, were honored with the Nobel Prize in Physiology and Medicine. Currently, vision studies at the Yerkes Regional Primate Research Center, for example, focus on myopia, post‐surgical treatment of congenital cataracts, adult cataract development and treatment, and laser surgery to correct corneal abnormalities. At the Wisconsin Regional Primate Research Center, the role of diet in visual impairment is evaluated during infancy. Diet is also studied to determine its role in the aging process, and the effects of aging on memory. Further, brain cells and other components of the nervous system in aging are also being investigated at these centers. © 1994 Wiley‐Liss, Inc.


title: Influence of Polyproline Region and Macro Domain Genetic Heterogeneity on HEV Persistence in Immunocompromised Patients

text: Hepatitis E virus (HEV) can chronically infect immunocompromised patients. The polyproline region (PPR) and the macro domain of ORF1 protein may modulate virus production and/or the host immune response. We investigated the association between the genetic heterogeneity of HEV quasispecies in ORF1 and the outcome of infection in solid-organ transplant patients. Both sequence entropy and genetic distances during the hepatitis E acute phase were higher in patients whose infection became chronic than in those who cleared the virus. Hence, great quasispecies heterogeneity in the regions encoding the PPR and the macro domain may facilitate HEV persistence.


title: Cdrom Archive: A Gateway to Study Camel Phenotypes

text: Camels are livestock that exhibit unique morphological, biochemical, and behavioral traits, which arose by natural and artificial selection. Investigating the molecular basis of camel traits has been limited by: (1) the absence of a comprehensive record of morphological trait variation (e.g., diseases) and the associated mode of inheritance, (2) the lack of extended pedigrees of specific trait(s), and (3) the long reproductive cycle of the camel, which makes the cost of establishing and maintaining a breeding colony (i.e., monitoring crosses) prohibitively high. Overcoming these challenges requires (1) detailed documentation of phenotypes/genetic diseases and their likely mode of inheritance (and collection of related DNA samples), (2) conducting association studies to identify phenotypes/genetic diseases causing genetic variants (instead of classical linkage analysis, which requires extended pedigrees), and (3) validating likely causative variants by screening a large number of camel samples from different populations. We attempt to address these issues by establishing a systematic way of collecting camel DNA samples, and associated phenotypic information, which we call the “Cdrom Archive.” Here, we outline the process of building this archive to introduce it to other camel researchers (as an example). Additionally, we discuss the use of this archive to study the phenotypic traits of Arabian Peninsula camel breeds (the “Mezayen” camels). Using the Cdrom Archive, we report variable phenotypic traits related to the coat (color, length, and texture), ear and tail lengths, along with other morphological measurements.


title: Ontology-driven aspect-based sentiment analysis classification: An infodemiological case study regarding infectious diseases in Latin America

text: Infodemiology is the process of mining unstructured and textual data so as to provide public health officials and policymakers with valuable information regarding public health. The appearance of this new data source, which was previously unimaginable, has opened up a new way in which to improve public health systems, resulting in better communication policies and better detection systems. However, the unstructured nature of the Internet, along with the complexity of the infectious disease domain, prevents the information extracted from being easily understood. Moreover, when dealing with languages other than English, for which some of the most common Natural Language Processing resources are not available, the correct exploitation of this data becomes even more difficult. We intend to fill these gaps proposing an ontology-driven aspect-based sentiment analysis with which to measure the general public’s opinions as regards infectious diseases when expressed in Spanish by employing a case study of tweets concerning the Zika, Dengue and Chikungunya viruses in Latin America. Our proposal is based on two technologies. We first use ontologies in order to model the infectious disease domain with concepts such as risks, symptoms, transmission methods or drugs, among other concepts. We then measure the relationship between these concepts in order to determine the degree to which one concept influences other concepts. This new information is subsequently applied in order to build an aspect-based sentiment analysis model based on statistical and linguistic features. This is done by applying deep-learning models. Our proposal is available on a web platform, where users can see the sentiment for each concept at a glance and analyse how each concept influences the sentiment of the others.


title: Relating structure to evolution in class II viral membrane fusion proteins

text: Enveloped viruses must fuse their lipid membrane to a cellular membrane to deliver the viral genome into the cytoplasm for replication. Viral envelope proteins catalyze this critical membrane fusion event. They fall into at least three distinct structural classes. Class II fusion proteins have a conserved three-domain architecture and are found in many important viral pathogens. Until 2013, class II proteins had only been found in flaviviruses and alphaviruses. However, in 2013 a class II fusion protein was discovered in the unrelated phlebovirus genus, and two unexpectedly divergent envelope proteins were identified in families that also contain prototypical class II proteins. The structural relationships of newly identified class II proteins, reviewed herein, shift the paradigm for how these proteins evolved.


title: Fabrication of Electrochemical-Based Bioelectronic Device and Biosensor Composed of Biomaterial-Nanomaterial Hybrid

text: The field of bioelectronics has paved the way for the development of biochips, biomedical devices, biosensors and biocomputation devices. Various biosensors and biomedical devices have been developed to commercialize laboratory products and transform them into industry products in the clinical, pharmaceutical, environmental fields. Recently, the electrochemical bioelectronic devices that mimicked the functionality of living organisms in nature were applied to the use of bioelectronics device and biosensors. In particular, the electrochemical-based bioelectronic devices and biosensors composed of biomolecule-nanoparticle hybrids have been proposed to generate new functionality as alternatives to silicon-based electronic computation devices, such as information storage, process, computations and detection. In this chapter, we described the recent progress of bioelectronic devices and biosensors based on biomaterial-nanomaterial hybrid.


title: Two Glycosylation Sites in H5N1 Influenza Virus Hemagglutinin That Affect Binding Preference by Computer-Based Analysis

text: Increasing numbers of H5N1 influenza viruses (IVs) are responsible for human deaths, especially in North Africa and Southeast Asian. The binding of hemagglutinin (HA) on the viral surface to host sialic acid (SA) receptors is a requisite step in the infection process. Phylogenetic analysis reveals that H5N1 viruses can be divided into 10 clades based on their HA sequences, with most human IVs centered from clade 1 and clade 2.1 to clade 2.3. Protein sequence alignment in various clades indicates the high conservation in the receptor-binding domains (RBDs) is essential for binding with the SA receptor. Two glycosylation sites, 158N and 169N, also participate in receptor recognition. In the present work, we attempted to construct a serial H5N1 HA models including diverse glycosylated HAs to simulate the binding process with various SA receptors in silico. As the SA-α-2,3-Gal and SA-α-2,6-Gal receptor adopted two distinctive topologies, straight and fishhook-like, respectively, the presence of N-glycans at 158N would decrease the affinity of HA for all of the receptors, particularly SA-α-2,6-Gal analogs. The steric clashes of the huge glycans shown at another glycosylation site, 169N, located on an adjacent HA monomer, would be more effective in preventing the binding of SA-α-2,3-Gal analogs.


title: A Murine Coronavirus MHV-S Isolate from Persistently infected Cells Has a Leader and Two Consensus Sequences between the M and N Genes

text: Abstract A plaque-cloned mouse hepatitis virus mutant, MHV-S No. 8, was isolated from Ki-BALB cells persistently infected with MHV-S. The mRNAs 1 to 6 were larger in the mutant, whereas there was no difference between the two viruses in the size of the smallest mRNA, mRNA 7. Sequence analyses of the genomic RNA, mRNA 6, and mRNA 7 of the two viruses revealed that an additional 111 nt were inserted just upstream of the intergenic consensus sequence preceding the N gene in MHV-S No. 8. The inserted region consisted of two different parts; the 3′-most 30 nt corresponded to nucleotides 28 to 57 of the leader sequence and the 5′-most 81 nt corresponded to nucleotides 58 to 138 of mRNA 7. This structure of No. 8 was most likely generated by RNA-RNA recombination between genomic RNA and subgenomic RNA species. The nucleotide insertion in the intergenic sequence between genes M and N resulted in two consensus sequences separated by 111 nt. Primer extension analysis revealed that the amount of a slightly larger, subgenomic mRNA resulting from initiation of synthesis at the upstream consensus sequence was only 5% of the usual sized mRNA 7 initiated from the downstream consensus sequence.


title: Hot topics en infecciones respiratorias

text: Resumen Se efectúa una revisión de los artículos más interesantes de infecciones respiratorias, publicados en el último trimestre del año 2009 y en 2010. En cuanto a las bronquiectasias, destacan las Guidelines de la British Thoracic Society, así como diversos artículos de la evolución natural del proceso y el impacto de las reagudizaciones de ésta, así como del tratamiento inhalatorio antibiótico de las bronquiectasias. Con respecto a la tuberculosis, es recomendable señalar el documento de consenso SEPAR-SEIMC para el manejo de la tuberculosis, así como las publicaciones sobre la utilización de interferón-gamma en el diagnóstico de infección tuberculosa. Las nuevas recomendaciones SEPAR de neumonía adquirida en la comunidad (NAC) se han publicado muy recientemente, y junto con ellas destacan estudios de etiología viral de las NAC, impacto del tratamiento corticoideo en las neumonías, duración del tratamiento antibiótico y medidas preventivas de neumonías, tanto comunitarias como nosocomiales. Abstract We review the most interesting articles on respiratory infections published in the last trimester of 2009 and in 2010. Notable publications in bronchiectasis were the Guidelines of the British Thoracic Society, as well as several articles on the natural course of the process, the impact of exacerbations on the course of the disease, and treatment with inhaled antibiotics. Other notable publications were the SEPAR-SEIMC consensus document for the management of tuberculosis and articles on the use of interferon-gamma in the diagnosis of tuberculosis infection. The new recommendations of the Spanish Society of Pneumology and Thoracic Surgery on community-acquired pneumonia have recently been published. Equally important are studies on the viral etiology of community-acquired pneumonia, the impact of corticosteroid treatment in pneumonia, the duration of antibiotic therapy and preventive measures in both community-acquired and nosocomial pneumonia.


title: Stage-Specific Requirement for Eomes in Mature NK Cell Homeostasis and Cytotoxicity

text: Natural killer (NK) cells are cytotoxic innate lymphoid cells (ILCs) that mediate antiviral and antitumor responses and require the transcriptional regulator Eomesodermin (Eomes) for early development. However, the role of Eomes and its molecular program in mature NK cell biology is unclear. To address this, we develop a tamoxifen-inducible, type-1-ILC-specific (Ncr1-targeted) cre mouse and combine this with Eomes-floxed mice. Eomes deletion after normal NK cell ontogeny results in a rapid loss of NK cells (but not ILC1s), with a particularly profound effect on penultimately mature stage III NK cells. Mechanisms responsible for stage III reduction include increased apoptosis and impaired maturation from stage II precursors. Induced Eomes deletion also decreases NK cell cytotoxicity and abrogates in vivo rejection of major histocompatibility complex (MHC)-class-I-deficient cells. However, other NK cell functional responses, and stage IV NK cells, are largely preserved. These data indicate that mature NK cells have distinct Eomes-dependent and -independent stages.


title: Relationship between mRNA stability and intron presence

text: Abstract Introns were found to enhance almost every steps of gene expression except increasing mRNA stability. By analyzing the genome-wide data of mRNA stability published by someone previously, we found that human intron-containing genes have more stable mRNAs than intronless genes, and the Arabidopsis thaliana genes with the most unstable mRNAs have fewer introns than other genes in the genome. After controlling for mRNA length, we found mRNA stability is still positively correlated with intron number in human intron-containing genes. But in yeast Saccharomyces cerevisiae, two different datasets on mRNA half-life gave conflicting results. The components of messenger ribonucleoprotein particles recruited during intron splicing may be retained in cytoplasmic mRNPs and act as signals of mRNA stability or simply insulators to avoid mRNA degradation.


title: Clinical Presentations for Influenza and Influenza-Like Illness in Young, Immunized Soldiers

text: Concern about respiratory diseases in soldiers increased in the late 1990s as production of the successful adenovirus vaccines stopped and the possibilities of an emergent pandemic influenza strain and use of bioweapons by terrorists were seriously considered. Current information on the causes and severity of influenza-like illness (ILI) was lacking. Viral agents and clinical presentations were described in a population of soldiers highly immunized for influenza. Using standard virus isolation techniques, 10 agents were identified in 164 (48.2%) of 340 soldiers hospitalized for ILI. Influenza isolates (29) and adenoviruses (98) occurred most frequently. Most influenza cases were caused by influenza A and probably resulted from a mismatch between circulating and vaccine viruses. Most (58.5%) patients with an adenovirus had a chest radiograph; 31.3% of these had an infiltrate. Clinical findings did not differentiate ILI caused by the various agents. Only 29 cases of influenza occurred in ∼7,200 person-years of observation, supporting the use of influenza vaccine.


title: Experimental studies of a coronavirus and coronavirus-like agent in a barrier-maintained feline breeding colony

text: Minimal-disease (MD) cats raised in a barrier-maintained feline breeding colony routinely become coronavirus (CV) antibody-positive at 5–8 weeks of age and remain seropositive indefinitely. In addition, they shed coronavirus-like particles (CVLPs) in their feces which are morphologically distinct from “typical” coronaviruses. Oronasal, intraperitoneal, intraduodenal, and intravenous inoculations of CVLPs were uniformly unsuccessful in the seroconversion of cats to CV, whereas subsequent prolonged close contact with colony cats did result in seroconversion. Such evidence indicates that MD cats in this barrier colony are infected with at least two non-cross-reactive agents: (1) feline enteric CVLPs, which are shed in feces and are morphologically and antigenically distinct from established coronaviruses (CVs), and (2) a virus which is antigenically cross-reactive with CVs, but which is apparently shed by some route other than the lower gastrointestinal tract.


title: Assessing ACE2 expression patterns in lung tissues in the pathogenesis of COVID-19

text: Abstract It has been reported that SARS-CoV-2 may use ACE2 as a receptor to gain entry into human cells, in a way similar to that of SARS-CoV. Analyzing the distribution and expression level of ACE2 may therefore help reveal underlying mechanisms of viral susceptibility and post-infection modulation. In this study, we utilized previously uploaded information on ACE2 expression in various conditions including SARS-CoA to evaluate the role of ACE2 in SARS-CoV and extrapolate that to COVID-19. We found that the expression of ACE2 in healthy populations and patients with underlying diseases was not significantly different. However, based on the elevated expression of ACE2 in cigarette smokers, we speculate that long-term smoking may be a risk factor for COVID-19. Analysis of ACE2 in SARS-CoV infected cells suggests that ACE2 is not only a receptor but is also involved in post-infection regulation, including immune response, cytokine secretion, and viral genome replication. Moreover, we constructed Protein-protein interaction (PPI) networks and identified hub genes in viral activity and cytokine secretion. Our findings may help clinicians and researchers gain more insight into the pathogenesis of SARS-CoV-2 and design therapeutic strategies for COVID-19.


title: 33 Viral Respiratory Infections

text: Abstract Acute respiratory illnesses are the most frequently occurring illness in all age groups globally. Disease is mostly limited to the upper airways and is self-limiting, but a small percentage can progress to lower respiratory tract infections as bronchiolitis and pneumonia. Children and elderly people are at increased risk, especially in developing countries. The most important etiologic agents of severe lower respiratory illness are bacteria such as Streptococcus pneumoniae and Haemophilus influenzae and viruses such as respiratory syncytial virus (RSV) and influenza virus. Efficacious vaccines are available against the two bacteria and the influenza virus. Viruses are much more important in mild upper and middle respiratory tract infections and in bronchiolitis in children, whereas bacteria are the main cause of pneumonia, especially in adults. Clinical syndromes overlap considerably, and there is increasing evidence of bacterial-viral co-infections and of bacterial pneumonia being secondary to viral respiratory tract infection.


title: COVID-19–associated acute pancreatitis: A rare cause of acute abdomen

text: Covid-19 is a viral disease that has spread throughout the world causing unprecedented impact on millions of people. It's a pandemic disease called officially by the World Health Organization (WHO) in March 11,2020. The disease mainly involves the upper respiratory tract system. Extrapulmonary manifestations are still not well understood. In this report, we describe a case of acute pancreatitis in a patient with Covid-19 infection who presented to our hospital.


title: A 6K-Deletion Variant of Salmonid Alphavirus Is Non-Viable but Can Be Rescued through RNA Recombination

text: Pancreas disease (PD) of Atlantic salmon is an emerging disease caused by Salmonid alphavirus (SAV) which mainly affects salmonid aquaculture in Western Europe. Although genome structure of SAV has been characterized and each individual viral protein has been identified, the role of 6K protein in viral replication and infectivity remains undefined. The 6K protein of alphaviruses is a small and hydrophobic protein which is involved in membrane permeabilization, protein processing and virus budding. Because these common features are shared across many viral species, they have been named viroporins. In the present study, we applied reverse genetics to generate SAV3 6K-deleted (Δ6K) variant and investigate the role of 6K protein. Our findings show that the 6K-deletion variant of salmonid alphavirus is non-viable. Despite viral proteins of Δ6K variant are detected in the cytoplasm by immunostaining, they are not found on the cell surface. Further, analysis of viral proteins produced in Δ6K cDNA clone transfected cells using radioimmunoprecipitation (RIPA) and western blot showed a protein band of larger size than E2 of wild-type SAV3. When Δ6K cDNA was co-transfected with SAV3 helper cDNA encoding the whole structural genes including 6K, the infectivity was rescued. The development of CPE after co-transfection and resolved genome sequence of rescued virus confirmed full-length viral genome being generated through RNA recombination. The discovery of the important role of the 6K protein in virus production provides a new possibility for the development of antiviral intervention which is highly needed to control SAV infection in salmonids.


title: COVID-19 and Diabetes: Knowledge in Progress

text: Abstract Aims We aimed to briefly review the general characteristics of the novel coronavirus (SARS-CoV-2) and provide a better understanding of the coronavirus disease (COVID-19) in people with diabetes, and its management. Methods We searched for articles in PubMed and Google Scholar databases till 02 April 2020, with the following keywords: “SARS-CoV-2”, “COVID-19”, “infection”, “pathogenesis”, “incubation period”, “transmission”, “clinical features”, “diagnosis”, “treatment”, “diabetes”, with interposition of the Boolean operator “AND”. Results The clinical spectrum of COVID-19 is heterogeneous, ranging from mild flu-like symptoms to acute respiratory distress syndrome, multiple organ failure and death. Older age, diabetes and other comorbidities are reported as significant predictors of morbidity and mortality. Chronic inflammation, increased coagulation activity, immune response impairment, and potential direct pancreatic damage by SARS-CoV-2 might be among the underlying mechanisms of the association between diabetes and COVID-19. No conclusive evidence exists to support the discontinuation of angiotensin-converting enzyme inhibitors (ACEI) or angiotensin receptor blockers because of COVID-19 in people with diabetes. Caution should be taken to potential hypoglycemic events with the use of chloroquine in these subjects. Patient tailored therapeutic strategies, rigorous glucose monitoring and careful consideration of drug interactions might reduce adverse outcomes. Conclusions Suggestions are made on the possible pathological mechanisms of the relationship between diabetes and COVID-19, and its management. No definite conclusions can be made based on current limited evidence. Further research regarding this relationship and its clinical management is warranted.


title: The economic impact of SARS: How does the reality match the predictions?

text: Abstract Objectives To perform a retrospective analysis of the macro-economic impact of the 2003 Severe Acute Respiratory Syndrome (SARS) outbreak. Methods As several years have now passed, it is possible to interrogate national statistics that have become available since the outbreak to provide a more accurate estimate of the actual macro-economic impact of SARS. National statistics were examined for anomalies that corresponded to the timing of the SARS outbreak and, where possible, the size of any gain or loss found estimated. Results Estimates and models produced at the time of the outbreak suggested that SARS could have a catastrophic effect on the global economy. Our analysis suggests that the scale of the SARS impact on affected economies was far smaller than suggested by contemporary media reports and model estimates. Conclusions This exercise holds important lessons for estimating the economic impact of future outbreaks – such as pandemic influenza – and measures to control or prevent them. We suggest that further work is needed to develop a more comprehensive macro-economic model able to more accurately estimate the relative cost and effect of a global response to outbreaks of international concern. The implications of our findings are discussed in the light of a prospective influenza pandemic.


The ongoing H1N1 flu pandemic and the intensive care community: challenges, opportunities, and the duties of scientific societies and intensivists


title: Evidence that multiple defects in murine DC-SIGN inhibit a functional interaction with pathogens

text: Abstract Certain viruses, bacteria, fungi and parasites target dendritic cells through the interaction with the cellular attachment factor DC-SIGN, making this C-type lectin an attractive target for therapeutic intervention. Studies on DC-SIGN function would be greatly aided by the establishment of a mouse model, however, it is unclear if the murine (m) homologue of human (h) DC-SIGN also binds to pathogens. Here, we investigated the interaction of mDC-SIGN, also termed CIRE, with the Ebolavirus glycoprotein (EBOV-GP), a ligand of hDC-SIGN. We found that mDC-SIGN neither binds EBOV-GP nor enhances infection by reporterviruses pseudotyped with EBOV-GP. Analysis of chimeras between mDC-SIGN and hDC-SIGN provided evidence that determinants in the carbohydrate recognition domain and in the neck domain of mDC-SIGN inhibit a functional interaction with EBOV-GP. Moreover, mDC-SIGN was found be monomeric, suggesting that lack of multimerization, which is believed to be required for efficient pathogen recognition by hDC-SIGN, might be one factor that prevents binding of mDC-SIGN to EBOV-GP. Our results suggest that mDC-SIGN on murine dendritic cells is not an adequate model for pathogen interactions with hDC-SIGN.


title: Public response to MERS-CoV in the Middle East: iPhone survey in six countries

text: Abstract Gulf Cooperation Council (GCC) countries bear the heaviest brunt of MERS-CoV. This study aims to compare public awareness and practice around MERS-CoV across GCC countries. A cross-sectional survey was conducted using the Gulf Indicators (GI) smartphone app among people in the six GCC countries, namely Saudi Arabia, Kuwait, the United Arab Emirates, Qatar, Bahrain, and Oman. A total of 1812 participants recruited. All were aware of MERS-CoV, yet the perception and practice around MERS-CoV varied widely between countries. Over two thirds were either “not concerned” or “slightly concerned” about contracting MERS-CoV; believing that they were under Allah’s (God’s) protection (40%) was the most cited reason. While 79% were aware that the disease can transmit through droplet from infected person, only 12% stated that MERS-CoV transmits via camels; people in Saudi Arabia were better aware of the transmission. Nevertheless, only 22% of respondents believed that camels are the zoonotic reservoir of MERS-CoV. Those who were concerned about contracting MERS-CoV (aOR: 1.6, 95% CI: 1.2–2.1, p <0.01) and those who thought MERS-CoV to be a severe disease only for those with high-risk conditions (aOR: 1.5, 95% CI: 1.1–2.1, p <0.01) were more likely to believe that camels are the zoonotic source. However, residents of KSA (aOR: 0.03, 95% CI: 0.01–0.07, p <0.01), UAE (aOR: 0.01, 95% CI: 0.004–0.02, p <0.01) and Kuwait (aOR: 0.03, 95% CI: 0.01–0.07, p <0.01) were less likely to believe that camels are the main zoonotic source compared to respondents from the other countries. Hygienic measures were more commonly adopted than avoidance of camels or their raw products, yet there was a discrepancy between the countries. This study highlights that despite being aware of the ongoing MERS-CoV epidemic; many people lack accurate understanding about MERS-CoV transmission, prevention, and are not fully compliant with preventive measures.


title: NINDS Strategies for Enhancing the Diversity of Neuroscience Researchers

text: Neuroscience is one of the fastest growing fields and highlights the excitement about research, but it also demonstrates the impact that our large scientific community can make in prioritizing equity and inclusion throughout science. I discuss strategies at multiple systemic levels where opportunities and interventions could be implemented to enhance neuroscience workforce diversity.


title: Ethical and Practical Considerations in Providing Critical Care to Patients With Ebola Virus Disease

text: Infectious disease epidemics in the past have given rise to psychologic and emotional responses among health-care workers (HCWs), stemming from fear of infection during patient care. Early experiences in the AIDS epidemic provide an example where fear of contagion resulted in differential treatment of patients infected with HIV. However, with a deeper understanding of AIDS pathogenesis and treatment, fear and discrimination diminished. Parallels exist between early experiences with AIDS and the present outbreak of Ebola virus disease in West Africa, particularly regarding discussions of medical futility in seriously ill patients. We provide a historical perspective on HCWs’ risk of infection during the provision of CPR, discuss physicians’ duty to treat in the face of perceived or actual HCW risk, and, finally, present the protocols implemented at the National Institutes of Health to reduce HCW risk while providing lifesaving and life-sustaining care.


title: Aerosolized avian influenza virus by laboratory manipulations

text: BACKGROUND: Avian H5N1 influenza viruses present a challenge in the laboratory environment, as they are difficult to collect from the air due to their small size and relatively low concentration. In an effort to generate effective methods of H5N1 air removal and ensure the safety of laboratory personnel, this study was designed to investigate the characteristics of aerosolized H5N1 produced by laboratory manipulations during research studies. RESULTS: Normal laboratory procedures used to process the influenza virus were carried out independently and the amount of virus polluting the on-site atmosphere was measured. In particular, zootomy, grinding, centrifugation, pipetting, magnetic stirring, egg inoculation, and experimental zoogenetic infection were performed. In addition, common accidents associated with each process were simulated, including breaking glass containers, syringe injection of influenza virus solution, and rupturing of centrifuge tubes. A micro-cluster sampling ambient air pollution collection device was used to collect air samples. The collected viruses were tested for activity by measuring their ability to induce hemagglutination with chicken red blood cells and to propagate in chicken embryos after direct inoculation, the latter being detected by reverse-transcription PCR and HA test. The results showed that the air samples from the normal centrifugal group and the negative-control group were negative, while all other groups were positive for H5N1. CONCLUSIONS: Our findings suggest that there are numerous sources of aerosols in laboratory operations involving H5N1. Thus, laboratory personnel should be aware of the exposure risk that accompanies routine procedures involved in H5N1 processing and take proactive measures to prevent accidental infection and decrease the risk of virus aerosol leakage beyond the laboratory.


title: From Touchdown to Transcription: The Reovirus Cell Entry Pathway

text: Mammalian orthoreoviruses (reoviruses) are prototype members of the Reoviridae family of nonenveloped viruses. Reoviruses contain ten double-stranded RNA gene segments enclosed in two concentric protein shells, outer capsid and core. These viruses serve as a versatile experimental system for studies of virus cell entry, innate immunity, and organ-specific disease. Reoviruses engage cells by binding to cell-surface carbohydrates and the immunoglobulin superfamily member, junctional adhesion molecule-A (JAM-A). JAM-A is a homodimer formed by extensive contacts between its N-terminal immunoglobulin-like domains. Reo-virus attachment protein σ1 disrupts the JAM-A dimer, engaging a single JAM-A molecule by virtually the same interface used for JAM-A homodimerization. Following attachment to JAM-A and carbohydrate, reovirus internalization is promoted by β1 integrins, most likely via clathrin-dependent endocytosis. In the endocytic compartment, reovirus outer-capsid protein σ3 is removed by cathepsin proteases, which exposes the viral membrane-penetration protein, μ1. Proteolytic processing and conformational rearrangements of μ1 mediate endosomal membrane rupture and delivery of transcriptionally active reovirus core particles into the host cell cytoplasm. These events also allow the ϕ cleavage fragment of μ1 to escape into the cytoplasm where it activates NF-κB and elicits apoptosis. This review will focus on mechanisms of reovirus cell entry and activation of innate immune response signaling pathways.


title: Expression of the B Subunit of E. coli Heat-labile Enterotoxin in the Chloroplasts of Plants and its Characterization

text: Transgenic chloroplasts have become attractive systems for heterologous gene expressions because of unique advantages. Here, we report a feasibility study for producing the nontoxic B subunit of Escherichia coli heat-labile enterotoxin (LTB) via chloroplast transformation of tobacco. Stable site-specific integration of the LTB gene into chloroplast genome was confirmed by PCR and genomic Southern blot analysis in transformed plants. Immunoblot analysis indicated that plant-derived LTB protein was oligomeric, and dissociated after boiling. Pentameric LTB molecules were the dominant molecular species in LTB isolated from transgenic tobacco leaf tissues. The amount of LTB protein detected in transplastomic tobacco leaf was approximately 2.5% of the total soluble plant protein, approximately 250-fold higher than in plants generated via nuclear transformation. The GM1–ELISA binding assay indicated that chloroplast-synthesized LTB protein bound to GM1-ganglioside receptors. LTB protein with biochemical properties identical to native LTB protein in the chloroplast of edible plants opens the way for inexpensive, safe, and effective plant-based edible vaccines for humans and animals.


title: Changing Trend of Infectious Diseases in Nepal

text: Many infectious/ communicable diseases (IDs) are endemic in Nepal. Until a decade and half ago, IDs were the major cause of both morbidity and mortality accounting 70% for both. However, as a result of various preventive measures implemented by both the state and non-state actors, the overall IDs have shown a changing (declining) trend. The most impressive decline has been seen in the intestinal helminth infection. Though the overall burden of IDs is decreasing, several newer infectious diseases (emerging infections) namely, dengue fever, scrub typhus, influenza (H5N1 and H1N1), and others are posing a great public health problem. On the other hand, though sporadic, outbreaks of endemic diseases together with HIV-TB coinfection and infection with drug resistance microbes during recent years have constituted a serious public health as well as medical problem. On the contrary, with the decline of IDs, noninfectious diseases (noncommunicable disease, NCD) namely, diabetes, cancer (and cancer therapy), and others are on the rise particularly in urban areas. Hence, currently Nepal is trapped in “double burden” of diseases. Risk of opportunistic infection has increased in immunocompromised person with NCD. To address the present situation, the multi-sectoral plan and strategies developed must be implemented effectively.


title: China’s response to a novel coronavirus stands in stark contrast to the 2002 SARS outbreak response

text: The strengthening of the Chinese Center for Disease Control and Prevention has been a turning point in outbreak responses in the area. This represents very welcome progress and development for global health security and diplomacy.


title: Performance of cows and summer-born calves and economics in semi-confined and confined beef systems

text: ABSTRACT Objectives Cow and calf performance in a semi-confined cow-calf production system was compared with total confinement using summer-born calves. Material and Methods The experiment was conducted over 3 yr in eastern Nebraska (ENREC) and 2 yr in western Nebraska (PREC) in a randomized complete block design. Lactating, crossbred beef cows (n = 127 at ENREC; n = 56 at PREC) with summer-born calves were used from November to April. Treatments were (1) dry-lot feeding or (2) cornstalk grazing with supplementation. Dry-lot pairs were limit fed a crop residue and distillers–based diet to meet energy requirements of a lactating cow. A dried distillers grain–based pellet was supplemented to pairs on cornstalks at a rate of 2.4 kg of DM/pair daily. Dry-lot cow-calf pairs were limit fed 12.1 kg of DM/d throughout the trial. Results and Discussion At ENREC, cows wintered on cornstalks lost BW and had a 0.46-unit decrease in BCS (P < 0.01), whereas cows in the dry-lot gained BW and had a 0.24-unit increase in BCS. At PREC, BCS increased by 0.03 units for cows wintered in the dry-lot and decreased by 0.26 units for cows wintered on cornstalks (P < 0.04). At both locations, calves fed in a dry-lot had greater ADG and BW per day of age compared with calves offered cornstalk grazing with supplementation (P ≤ 0.03). Implications and Applications A partial budget suggests that lower winter production inputs compensate for reduced performance of calves when cow-calf pairs are wintered on cornstalks, making residue grazing a viable option in partial confinement systems.


title: Interferons: Reprogramming the Metabolic Network against Viral Infection

text: Viruses exploit the host and induce drastic metabolic changes to ensure an optimal environment for replication and the production of viral progenies. In response, the host has developed diverse countermeasures to sense and limit these alterations to combat viral infection. One such host mechanism is through interferon signaling. Interferons are cytokines that enhances the transcription of hundreds of interferon-stimulated genes (ISGs) whose products are key players in the innate immune response to viral infection. In addition to their direct targeting of viral components, interferons and ISGs exert profound effects on cellular metabolism. Recent studies have started to illuminate on the specific role of interferon in rewiring cellular metabolism to activate immune cells and limit viral infection. This review reflects on our current understanding of the complex networking that occurs between the virus and host at the interface of cellular metabolism, with a focus on the ISGs in particular, cholesterol-25-hydroxylase (CH25H), spermidine/spermine acetyltransferase 1 (SAT1), indoleamine-2,3-dioxygenase (IDO1) and sterile alpha motif and histidine/aspartic acid domain-containing protein 1 (SAMHD1), which were recently discovered to modulate specific metabolic events and consequently deter viral infection.


title: Evolution of infectious bronchitis virus in Taiwan: Characterisation of RNA recombination in the nucleocapsid gene

text: Abstract Avian infectious bronchitis virus (IBV) belongs to the Coronaviridae family and causes significant economic loss in Taiwan (TW), even in flocks that have been extensively immunised with Massachusetts (Mass)-serotype vaccines. Phylogenetic analysis of all non-structural and most structural genes shows that TW IBV is genetically distinct from the US strain and more similar to Chinese (CH) IBV. In contrast, the nucleocapsid (N) gene of TW IBV presents phylogenetic incongruence. RNA recombination at the 5′ end of the N gene between TW and US IBV is shown to be responsible for this discordance. Surprisingly, the recombinant N gene is found in all of tested TW IBV isolates, suggesting that a recombination event gave origin to a founder lineage. Our data indicate that RNA recombination in the recombinant 5′ end of the N gene may have caused the emergence of the current IBV population in Taiwan.


title: Significance of crinoid preservation: Clare Shale Formation (Upper Carboniferous), Fisherstreet Bay, Doolin, County Clare, Ireland

text: Some fossils, such as crinoid stems, are not widely appreciated by collectors and researchers, yet can provide unique data regarding taphonomy and palaeoecology. A long crinoid pluricolumnal showing a distinctive pattern of preservation was collected from the Clare Shale Formation (Upper Carboniferous) at Fisherstreet Bay, Doolin, County Clare, western Ireland. The specimen is partly disarticulated and represents the mesistele to mesistele/dististele transition; attachment was by unbranched long, slender radices; pluricolumnal heteromorphic; fragments of pluricolumnal are of multiples of a unit length. This specimen, cladid? sp. indet., slumped to the seafloor after death and started to disarticulate as ligaments rotted. By reference to the broken stick model, the pattern of disarticulation suggests that the noditaxis of the heteromorphic stem was N212.


title: Analysis of the spike, ORF3, and nucleocapsid genes of porcine epidemic diarrhea virus circulating on Thai swine farms, 2011–2016

text: Porcine epidemic diarrhea virus (PEDV) outbreaks on pig farms have caused significant economic loss in the swine industry since it was first reported in Thailand a decade ago. Anecdotal evidence suggests that PEDV is now endemic in this region, therefore genome information of circulating PEDV is important for molecular surveillance and evaluation of potential benefits of field vaccination. Here, we characterized PEDV infection on commercial Thai swine farms by screening 769 samples of feces and small intestinal contents from pigs with diarrhea between 2011 and 2016. Using reverse-transcription polymerase chain reaction targeting the spike (S) gene, 153 PEDV-positive samples were further subjected to analysis of the open reading frame 3 and nucleocapsid (N) genes. Comparison of 95 samples in which nucleotide sequencing was successfully obtained for all three genes revealed evolutionary diversity among the Thai PEDV strains. Phylogenetic analyses suggest that although some Thai strains changed little from years past, others resembled more closely to the recent strains reported in China. Interestingly, eight Thai PEDV strains possessed amino acid deletions in the N protein. The PEDV sequence divergence may be responsible for driving periodic outbreaks and continued persistence of PEDV on commercial swine farms. Our findings provide important insight into regional PEDV strains in circulation, which may assist future inclusions of suitable strains for future PEDV vaccines.


title: Point of care procedures in physiatry: Practice considerations during the Covid-19 pandemic

text: Coronavirus Disease 2019 (Covid-19) is an active pandemic that has required rapid conversion of practice patterns to mitigate disease spread. Although recommendations have been released for physicians to postpone elective procedures, the utility of common physiatry procedures and their infectious risk profile have yet to be clearly delineated. In this article, we describe an update on existing national recommendations and outline considerations as practitioners and institutions strive to meet the needs of patients with disabilities.


title: Vectors for gene expression in mammalian cells

text: Publisher Summary The achievement of robust and regulated protein production in mammalian cells is a complex process that requires careful consideration of many factors, including transcriptional and translational control elements, RNA processing, gene copy number, mRNA stability, the chromosomal site of gene integration, potential toxicity of recombinant proteins to the host cell, and the genetic properties of the host. Gene transfer into mammalian cells may be effected either by infection with virus that carries the recombinant gene of interest, or by direct transfer of plasmid DNA. This chapter discusses the molecular architecture of non-viral vectors for high-level protein production. Virus-based vectors for gene therapy, protein production, vaccine development and other applications are summarized in a table and described.


Your liberty or your life. Talking Point on public health versus civil liberties


title: Can video consultations replace face-to-face interviews? Palliative medicine and the Covid-19 pandemic: rapid review

text: During the Covid-19 pandemic, a strategy to minimise face-to-face (FtF) visits and limit viral spread is essential. Video consultations offer clinical assessment despite restricted movement of people. We undertook a rapid literature review to identify the highest currently available level of evidence to inform this major change in clinical practice. We present a narrative synthesis of the one international and one national guideline and two systematic reviews—all published within the last 18 months. The global evidence appears to support video consultations as an effective, accessible, acceptable and cost-effective method of service delivery. Organisations must ensure software is simple, effective, reliable and safe, with the highest level of security for confidentiality. Although video consultations cannot fully replace FtF, they can radically reduce the need for FtF and the risk of Covid-19 spread in our communities while maintaining high standards of care. For patient safety, it will be critical to follow the WHO guidance regarding: a standard operating procedure; clinical protocols for when video consultations can (and cannot) be used; policies to ensure equity of access in disadvantaged populations; adequate staff training; and administrative support to coordinate appointments.


title: Detection of Viral RNA Fragments in Human iPSC-Cardiomyocytes following Treatment with Extracellular Vesicles from SARS-CoV-2 Coding-Sequence-Overexpressing Lung Epithelial Cells

text: The novel coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has evolved into a worldwide pandemic. Early data suggest that the prevalence and severity of COVID-19 appear to be higher among patients with underlying cardiovascular risk factors. Despite the expression of angiotensin-converting enzyme 2 (ACE2), a functional receptor for SARS-CoV-2 infection, in cardiomyocytes, there has been no conclusive evidence of direct viral infection although the presence of inflammation and viral genome within the hearts of COVID-19 patients have been reported. Here we transduced A549 lung epithelial cells with lentivirus overexpressing selected genes of the SARS-CoV-2. We then isolated extracellular vesicles (EVs) from the supernatant of A549 cells and detected the presence of viral RNA within the purified EVs. Importantly, we observed that human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were able to actively uptake these EVs and viral genes were subsequently detected in the cardiomyocytes. Accordingly, uptake of EVs containing viral genes led to an upregulation of inflammation-related genes in hiPSC-CMs. Thus, our findings indicate that SARS-CoV-2 RNA-containing EVs represent an indirect route of viral RNA entry into cardiomyocytes.


title: Leptomeningitis and polycaryocyte formation in the CNS of rats inoculated subcutaneously with Herpes simplex virus

text: Abstract Wistar rats were inoculated subcutaneously with either type 1 (HSV1) or type 2 (HSV2) Herpes simplex virus at 5 days of age. Animals were killed in extremis or at the end of the 14-day observation period postinoculation. Acute destructive meningoencephalitis with hemorrhage and leukocytic infiltration was observed in both groups. Polycaryocytes comprised of cells of the internal granular layer of the cerebellum were observed in some animals inoculated with HSV1. These multinucleated cells appeared to be formed by fusion of virus-infected cells, and intranuclear inclusion bodies were observed. Lesions in the leptomeninges were particularly striking in animals inoculated with HSV2. Viral replication in resident cells of the leptomeninges was demonstrated by electron microscopy.


A Primary Care Response to COVID‐19 for Patients with an Opioid Use Disorder


title: Nitric oxide is elicited and inhibits viral replication in pigs infected with porcine respiratory coronavirus but not porcine reproductive and respiratory syndrome virus

text: There is little information on the role of nitric oxide ((•)NO) in innate immunity to respiratory coronavirus (CoV) infections. We examined (•)NO levels by Greiss assay in bronchoalveolar lavage (BAL) of pigs infected with either porcine respiratory coronavirus (PRCV) or porcine reproductive and respiratory syndrome virus (PRRSV), a member of Nidovirales, like CoV. The antiviral effects of (•)NO on these two viruses were tested in an in vitro system using a (•)NO donor, S-nitroso-N-acetylpenicillamine (SNAP). We detected a large increase in (•)NO levels in BAL fluids of PRCV-infected pigs, but not in PRRSV-infected pigs. Pulmonary epithelial cell necrosis induced by PRCV coincided with increased (•)NO. Moreover, (•)NO levels in cell culture medium of PRRSV-infected alveolar macrophages (AMs) did not differ from that of mock-infected AMs. Antiviral assays showed that (•)NO significantly inhibited PRCV replication in swine testicular (ST) cells, whereas PRRSV was not susceptible to (•)NO based on the conditions tested. Our study suggests that unlike PRRSV which induces apoptosis in AMs, respiratory CoVs such as PRCV that infect pulmonary epithelial cells and cause cytolysis, induce (•)NO production in the respiratory tract. Thus, (•)NO may play a role in innate immunity to respiratory CoV infections by inhibiting viral replication.


COVID-19 and the Role of Spine Surgeons.


title: Chapter 8 Environmental Factors and Their Regulation of Immunity in Multiple Sclerosis

text: Abstract In multiple sclerosis (MS), environmental factors and genetic traits cooperate in the induction of the chronic activation of immune cells to produce the brain pathology. Epidemiology has focused on different environmental risk factors but certainly virus infection, smoking, vitamin D levels, and sunlight exposure are the most relevant. What is certainly less clear is the way in which these external factors are able to induce and sustain the internal pathology process of the disease. Epigenetics has been recently focused on trying to shed light on this aspect. As a matter of fact epigenetic changes are highly sensitive to environmental factors that therefore may influence the susceptibility to the disease by acting through epigenetic modifications. In this chapter we discuss the most relevant environmental factors and how they may affect the immune response in MS. Finally, we discuss the possible role of the microbiota in inducing autoimmunity in MS.


title: Chitosan-based mucosal adjuvants: Sunrise on the ocean

text: Mucosal vaccination, which is shown to elicit systemic and mucosal immune responses, serves as a non-invasive and convenient alternative to parenteral administration, with stronger capability in combatting diseases at the site of entry. The exploration of potent mucosal adjuvants is emerging as a significant area, based on the continued necessity to amplify the immune responses to a wide array of antigens that are poorly immunogenic at the mucosal sites. As one of the inspirations from the ocean, chitosan-based mucosal adjuvants have been developed with unique advantages, such as, ability of mucosal adhesion, distinct trait of opening the junctions to allow the paracellular transport of antigen, good tolerability and biocompatibility, which guaranteed the great potential in capitalizing on their application in human clinical trials. In this review, the state of art of chitosan and its derivatives as mucosal adjuvants, including thermo-sensitive chitosan system as mucosal adjuvant that were newly developed by author's group, was described, as well as the clinical application perspective. After a brief introduction of mucosal adjuvants, chitosan and its derivatives as robust immune potentiator were discussed in detail and depth, in regard to the metabolism, safety profile, mode of actions and preclinical and clinical applications, which may shed light on the massive clinical application of chitosan as mucosal adjuvant.


title: Controlling SARS: a review on China’s response compared with other SARS‐affected countries

text: Objective To summarise the major control measures implemented by severe acute respiratory syndrome (SARS)‐affected countries and to compare distinctive features of the Chinese approach to other affected Asian countries and Canada. Method Literature review. Results The realisation in March 2003 that SARS was spreading led affected countries to introduce measures such as rapid dissemination of information, early case detection and isolation, tracing and quarantining of SARS contacts, traveller screening, raising public awareness of risk and institution of stricter infection control in health care settings. SARS became a notifiable disease in China in mid‐April 2003, after which introduction of efficient nationwide control measures led to containment within 2 months. Countries differed in the timeliness of implementing control measures, the mode and extent to which these were enforced and in the resources available to do so. Conclusion SARS challenged the political and public health systems of all affected countries. It demanded rapid and decisive action to be taken, yet the comparison shows how difficult this was for an unknown new disease. Guangdong reacted rapidly but this pace was not continued by China for some time, which facilitated national and international spread. Once the Chinese government changed its policy, it developed an impressive control strategy involving the public which culminated in containment. The significance of timely information was perhaps the main lesson which the SARS epidemic taught.


title: Role of the lipid rafts in the life cycle of canine coronavirus

text: Coronaviruses are enveloped RNA viruses that have evolved complex relationships with their host cells, and modulate their lipid composition, lipid synthesis and signalling. Lipid rafts, enriched in sphingolipids, cholesterol and associated proteins, are special plasma membrane microdomains involved in several processes in viral infections. The extraction of cholesterol leads to disorganization of lipid microdomains and to dissociation of proteins bound to lipid rafts. Because cholesterol-rich microdomains appear to be a general feature of the entry mechanism of non-eneveloped viruses and of several coronaviruses, the purpose of this study was to analyse the contribution of lipids to the infectivity of canine coronavirus (CCoV). The CCoV life cycle is closely connected to plasma membrane cholesterol, from cell entry to viral particle production. The methyl-β-cyclodextrin (MβCD) was employed to remove cholesterol and to disrupt the lipid rafts. Cholesterol depletion from the cell membrane resulted in a dose-dependent reduction, but not abolishment, of virus infectivity, and at a concentration of 15 mM, the reduction in the infection rate was about 68 %. MβCD treatment was used to verify if cholesterol in the envelope was required for CCoV infection. This resulted in a dose-dependent inhibitory effect, and at a concentration of 9 mM MβCD, infectivity was reduced by about 73 %. Since viral entry would constitute a target for antiviral strategies, inhibitory molecules interacting with viral and/or cell membranes, or interfering with lipid metabolism, may have strong antiviral potential. It will be interesting in the future to analyse the membrane microdomains in the CCoV envelope.


The COVID-19 global inflection point and Europe’s predicament


title: Chapter 05 Therapeutic areas Strategically important diseases of the future

text: Abstract Society and industry—still the only provider of approved medicine—share the desire to treat important diseases. Governments too, since some diseases’ high prevalence affect the economic output of society. Yet the decisions as to which drugs will be developed are strictly in private hands. The risks required to treat some really important, and often increasingly prevalent, diseases mean that the desires of society and the industry are diverging. This is not boding well for our society. The understanding and willingness of governments to step in as a partner and financier is not apparent. Will governments recognize their role in this? Since the economic cost of these diseases, in terms of reduced productivity and medical costs to treat the disease, is enormous, understanding should be forthcoming. Government has invested for many decades in an enormous program of cancer research, which now shows dividends. The importance of further government investment in basic research is apparent when industry does not want to or cannot anymore afford it. As Big Pharma drops out of important therapeutic areas, it is not promising to assume that government can step in to the vacancy. There has to be a serious effort to find a formula by which the government gently or not so gently forces the industry to continue its efforts because it is so important. Perhaps the most important of these neglected diseases is Alzheimer’s disease (AD) because of its dehumanizing nature and its predictably massively increasing prevalence. Yet, none of the currently approved Alzheimer drugs can be used to prevent, cure, or even significantly slow the progression of this disease. It is predicted that by 2030 50% of the over 80s will have the disease. Almost 100% of people with an inheritable trait will develop the disease unless we succeed in finding a preventive therapy. Only recently has a preventive trial been announced. It is often thought that insulin-dependent diabetes is a well-treated disease and needs no further diagnostic and drug development efforts. This could not be further from the truth. The long-term complications of insulin-treated diabetes add to the economic burden, especially as the growth in prevalence of the disease expands with the waistlines of the world’s population. Three hundred million may have type 2 diabetes by 2030. Pharmaceutical control of weight loss has so far been vexed by serious side effects. When it comes to diabetes drugs, we start to recognize that the control of blood glucose can be further improved; the biggest contribution is not by drugs, but by self-diagnosis with point of care glucose measuring devices. Morphine is still the most effective medicine against pain. It has been around for 5,000 years and we haven’t come up with anything better. But it can be ineffective against neuropathic pain and cancer pain. New pain medicines are difficult to find because of the subjective measurement and poor animal models. Since many current pain killers are abused, there are other hurdles. Despite the many difficulties, society needs new pain killers at the same time that notable pharma are pulling out, despite the economic opportunity. Drugs against cancer have had many high-profile and expensive failures in recent years, with all major companies having at least one failed candidate. Biotech companies have made much progress, however, and they are either collaborating with or being taken over by Big Pharma. Oncology drug development has been crucially enabled by decades of government investment. The key is the many characterized drug targets that other disease areas do not have. Government intervention in oncology and HIV provide the model for treatment of AD, neuropathic pain, and schizophrenia, among other therapeutic areas. Increasing governmental direct funding of translational research is not the solution many hope it will be.


title: Targeting regenerative exosomes to myocardial infarction using cardiac homing peptide

text: Rationale: Cardiac stem cell-derived exosomes have been demonstrated to promote cardiac regeneration following myocardial infarction in preclinical studies. Recent studies have used intramyocardial injection in order to concentrate exosomes in the infarct. Though effective in a research setting, this method is not clinically appealing due to its invasive nature. We propose the use of a targeting peptide, cardiac homing peptide (CHP), to target intravenously-infused exosomes to the infarcted heart. Methods: Exosomes were conjugated with CHP through a DOPE-NHS linker. Ex vivo targeting was analyzed by incubating organ sections with the CHP exosomes and analyzing with fluorescence microscopy. In vitro assays were performed on neonatal rat cardiomyocytes and H9C2 cells. For the animal study, we utilized an ischemia/reperfusion rat model. Animals were treated with either saline, scramble peptide exosomes, or CHP exosomes 24 h after surgery. Echocardiography was performed 4 h after surgery and 21 d after surgery. At 21 d, animals were sacrificed, and organs were collected for analysis. Results: By conjugating the exosomes with CHP, we demonstrate increased retention of the exosomes within heart sections ex vivo and in vitro with neonatal rat cardiomyocytes. In vitro studies showed improved viability, reduced apoptosis and increased exosome uptake when using CHP-XOs. Using an animal model of ischemia/reperfusion injury, we measured the heart function, infarct size, cellular proliferation, and angiogenesis, with improved outcomes with the CHP exosomes. Conclusions: Our results demonstrate a novel method for increasing delivery of for treatment of myocardial infarction. By targeting exosomes to the infarcted heart, there was a significant improvement in outcomes with reduced fibrosis and scar size, and increased cellular proliferation and angiogenesis.


title: Puzzle over viral load

text: We don't yet know if being exposed to more coronavirus particles leads to more severe covid-19 symptoms, reports Linda Geddes


title: Mapping and characterization of G-quadruplexes in Mycobacterium tuberculosis gene promoter regions

text: Mycobacterium tuberculosis is the causative agent of tuberculosis (TB), one of the top 10 causes of death worldwide in 2015. The recent emergence of strains resistant to all current drugs urges the development of compounds with new mechanisms of action. G-quadruplexes are nucleic acids secondary structures that may form in G-rich regions to epigenetically regulate cellular functions. Here we implemented a computational tool to scan the presence of putative G-quadruplex forming sequences in the genome of Mycobacterium tuberculosis and analyse their association to transcription start sites. We found that the most stable G-quadruplexes were in the promoter region of genes belonging to definite functional categories. Actual G-quadruplex folding of four selected sequences was assessed by biophysical and biomolecular techniques: all molecules formed stable G-quadruplexes, which were further stabilized by two G-quadruplex ligands. These compounds inhibited Mycobacterium tuberculosis growth with minimal inhibitory concentrations in the low micromolar range. These data support formation of Mycobacterium tuberculosis G-quadruplexes in vivo and their potential regulation of gene transcription, and prompt the use of G4 ligands to develop original antitubercular agents.


Applying (or not?) CAR-T Neurotoxicity Experience to COVID19 Delirium and Agitation


title: “Challenges for liver transplantation during recovery from the COVID-19 pandemic: insights and recommendations”

text: Abstract The coronavirus disease 2019 (COVID-19) pandemic is placing an increasing burden on liver transplant (LT) services worldwide. At the peak of the pandemic, many liver transplant (LT) services worldwide dropped down or halted their activities. With the gradual easing of lockdowns, LT teams face new challenges when restarting activities. The numbers of liver transplants are likely to drop in the immediate post-COVID era. Prolong, and intermittent lockdowns are likely to lead to a shortage of supplies, especially in poor resource settings. Special attention is needed to avoid nosocomial COVID-19 infection among cirrhotic patients awaiting transplant, post-transplant patients as well as members in transplant teams. LT programs may have to revise the existing strategies in selecting donors and recipients for transplants. Re-designing service provision, restructuring of outpatient care, careful screening and selection of donors and recipient, performing LT with limited resources will have to be initiated in the post-COVID era if long-term recovery of LT services are to be expected. Costs involved with LT are likely to increase, considering the change in protocols of testing, quarantining, and interstate traveling. This paper discusses the different elements affecting and the widespread impact of COVID-19 pandemic on LT and strategies to minimize the impact of these factors and to adapt so LT services can meet the healthcare needs during this pandemic and beyond.