title: Safety Assessment of a Hemp Extract using Genotoxicity and Oral Repeat-Dose Toxicity Studies in Sprague-Dawley Rats

text: Cannabinoids are extracted from Cannabis sativa L. and are used for a variety of medicinal purposes. Recently, there has been a focus on the cannabinoid Cannabidiol (CBD) and its potential benefits. This study investigated the safety of a proprietary extract of C. sativa, consisting of 9% hemp extract (of which 6.27% is CBD) and 91% olive oil. The mutagenic potential of the hemp extract was evaluated with the AMES assay inclusive of a hepatic drug metabolizing mix (S9) rich in CYP enzymes. The test article did not elicit evidence of bacterial mutagenicity. GLP compliant 14-day and a 90-day toxicity study were conducted. Olive oil was used as a control. The 90-day study had a 28-day recovery period. Treatments for the 14-day non-recovery range-finding study were 0, 1000, 2000 and 4000 mg test article/kg body weight (bw)/day for 14 days. There was a non-statistically significant (p > 0.05) decrease in body weights for the male and female rats receiving the test article. Hypoactivity, hyperactivity, reduced food consumption and piloerection were observed in the rats receiving 4000 mg test article/kg bw. Histopathology showed an increase in the size of liver cells (hypertrophy) around the central vein (centrilobular) in Groups 3 (3/10) and 4 (5/10) that correlated with increased liver weights. In the 90-day study, 8 groups of rats were dosed with 0, 200, 400 and 800 mg test article/kg bw/day. Groups 5 to 8 had a 28-day recovery. There were no test article-linked changes in clinical observations, physical examinations, Functional Observation Battery, ophthalmology, Motor Activity Assessment, hematology, clinical chemistries and macropathology (all groups). With the exception of the liver and adrenal gland, no test article-linked pathology was observed. For all rats receiving the test article, histopathology showed hypertrophy of liver cells around the central vein. The increase of liver weight is most likely caused by hypertrophy due to up-regulation of the hepatic drug metabolizing enzymes. The hepatocellular hypertrophy was completely reversed in 28 days and was not considered to be an adverse effect. Vacuolization of the adrenal zona fasciculata was observed in the control and 800 mg test article/kg bw groups. The vacuolization of the zona fasciculata was of the same incidence and severity in treatment and control male rats and correlated with an increased in the weights of the adrenal glands. In addition, a statistically significant increase (p<0.05) in adrenal-to-body weight ratios was observed for females receiving 800 mg test article/kg bw. This increase in adrenal-to-body weight ratio did not correlate with any of the pathology findings. The NOAEL for the test article is 800 mg/kg bw/day for female and 400 mg/kg bw/day for male Sprague Dawley rats.


title: Effective Reproductive Number estimation for initial stage of COVID-19 pandemic in Latin American Countries

text: Abstract Objectives The coronavirus disease 2019 (COVID-19) has become pandemic and turn in a challenge for Latin America. Understanding the dynamics of the epidemic is essential for decision making, and to reduce the health, economic, and social impacts of the pandemic. The present study aimed to estimate the effective reproductive number (Rt) of Severe Acute Respiratory Syndrome coronavirus 2 (SARS-Cov2) infection during the first 10 days of the outbreak in seven Latin American countries with the highest incidence of cases as of March 23, 2020. Furthermore, we chose to compare the seven countries with Spain and Italy given their history with the virus. Methods Incidence data retrieved from the COVID-19 data repository by Johns Hopkins University were analyzed. The Rt was calculated for the first 10 days of the epidemic in Brazil, Ecuador, Chile, Colombia, Panama, Mexico, and Peru. Rt estimations were compared with Spain and Italy values for the same interval. Results The median Rt for the first 10 days of the COVID-19 epidemic were 2.90 (2.67-3.14) for Spain and 2.83 (2.7-2.96) for Italy. Latin American Rt estimations were higher in Ecuador (3.95(3.7-4.21)), Panama (3.95(3.7-4.21)), and Brazil (3.95(3.7-4.21)). The smallest one was observed in Peru (2.36(2.11-2.63)). All Latin American countries had Rt greater than 2. Conclusions The initial stages of the COVID-19 epidemic in Latin America suggested a high Rt. Interventions such as domestic and international travel restrictions, educational institutions closure, social distancing, and intensified case surveillance should be adopted to prevent the collapse of the health systems.


title: G-quadruplex stabilization in the ions and maltose transporters inhibit Salmonella enterica growth and virulence

text: The G-quadruplex structure forming motifs have recently emerged as a novel therapeutic drug target in various human pathogens. Herein, we report three highly conserved G-quadruplex motifs (SE-PGQ-1, 2, and3) in genome of all the 412 strains of Salmonella enterica. Bioinformatics analysis inferred the presence of SE-PGQ-1 in the regulatory region of mgtA, presence of SE-PGQ-2 in the open reading frame of entA and presence of SE-PGQ-3 in the promoter region of malE and malK genes. The products of mgtA and entA are involved in transport and homeostasis of Mg2+ and Fe3+ ion and thereby required for bacterial survival in the presence of reactive nitrogen/oxygen species produced by the host macrophages, whereas, malK and malE genes are involved in transport of maltose sugar, that is one of the major carbon source in the gastrointestinal tract of human. The formation of stable intramolecular G-quadruplex structures by SE-PGQs was confirmed by employing CD, EMSA and NMR spectroscopy. Cellular studies revealed the inhibitory effect of 9-amino acridine on Salmonella enterica growth. Next, CD melting analysis demonstrated the stabilizing effect of 9-amino acridine on SE-PGQs. Further, polymerase inhibition and RT-qPCR assays emphasize the biological relevance of predicted G-quadruplex in the expression of PGQ possessing genes and demonstrate the G-quadruplexes as a potential drug target for the devolping novel therapeutics for combating Salmonella enterica infection. Author Summary Since last several decades’ scientific community has witnessed a rapid increase in number of such human pathogenic bacterial species that acquired resistant to multiple antibacterial agents. Currently, emergence of multidrug-resistant strains remain a major public health concern for clinical investigators that rings a global alarm to search for novel and highly conserved drug targets. Recently, G-quadruplex structure forming nucleic acid sequences were endorsed as highly conserved Drug target for preventing infection of several human pathogens including viral and protozoan species. Therefore, here we explored the presence G-quadruplex forming motif in genome of Salmonella enterica bacteria that causes food poisoning, and enteric fever in human. The formation of intra molecular G-quadruplex structure in four genes (mgtA, entA, malE and malK) was confirmed by NMR, CD and EMSA. The 9-amino acridine, a known G-quadruplex binder has been shown to stabilize the predicted G-quadruplex motif and decreases the expressioin of G-quadruplex hourbouring genes using RT-PCR and cellular toxicity assay. This study concludes the presence of G-quadruplex motifs in essential genes of Salmonella enterica genome as a novel and conserved drug target and 9-amino acridine as candidate small molecule for preventing the infection of Salmonella enterica using a G4 mediated inhibition mechanism.


title: Gastroenteritis viruses: research update and perspectives

text: Abstract Gastroenteritis Viruses, Novartis Foundation Symposium 238 London, UK, 16–18 May 2000


title: Longitudinal molecular microbial analysis of influenza-like illness in New York City, may 2009 through may 2010

text: BACKGROUND: We performed a longitudinal study of viral etiology in samples collected in New York City during May 2009 to May 2010 from outpatients with fever or respiratory disease symptoms in the context of a pilot respiratory virus surveillance system. METHODS: Samples were assessed for the presence of 13 viruses, including influenza A virus, by MassTag PCR. RESULTS: At least one virus was detected in 52% of 940 samples analyzed, with 3% showing co-infections. The most frequently detected agents were rhinoviruses and influenza A, all representing the 2009 pandemic H1N1 strain. The incidence of influenza H1N1-positive samples was highest in late spring 2009, followed by a decline in summer and early fall, when rhinovirus infections became predominant before H1N1 reemerged in winter. Our study also identified a focal outbreak of enterovirus 68 in the early fall of 2009. CONCLUSION: MassTag multiplex PCR affords opportunities to track the epidemiology of infectious diseases and may guide clinicians and public health practitioners in influenza-like illness and outbreak management. Nonetheless, a substantial proportion of influenza-like illness remains unexplained underscoring the need for additional platforms.


title: Current therapy for chronic hepatitis C: The role of direct-acting antivirals

text: Abstract One of the most exciting developments in antiviral research has been the discovery of the direct-acting antivirals (DAAs) that effectively cure chronic hepatitis C virus (HCV) infections. Based on more than 100 clinical trials and real-world studies, we provide a comprehensive overview of FDA-approved therapies and newly discovered anti-HCV agents with a special focus on drug efficacy, mechanisms of action, and safety. We show that HCV drug development has advanced in multiple aspects: (i) interferon-based regimens were replaced by interferon-free regimens; (ii) genotype-specific drugs evolved to drugs for all HCV genotypes; (iii) therapies based upon multiple pills per day were simplified to a single pill per day; (iv) drug potency increased from moderate (∼60%) to high (>90%) levels of sustained virologic responses; (v) treatment durations were shortened from 48 to 12 or 8 weeks; and (vi) therapies could be administered orally regardless of prior treatment history and cirrhotic status. However, despite these remarkable achievements made in HCV drug discovery, challenges remain in the management of difficult-to-treat patients.


title: Pandemics: is hoping for the best enough?

text: Albert Osterhaus reflects on the reasons for the increase in emerging pandemics and how we are coping with these situations. Although we might risk over-reacting, better safe than sorry is still the most reasonable policy.


title: COVID- 19 un Syndrome de Détresse Respiratoire Aigu atypique

text: Résumé L’infection au nouveau coronavirus Sars- Cov 2 est responsable d’une forme sévère de pneumonie, la COVID-19 pouvant évoluer vers un tableau de syndrome de détresse respiratoire aiguë. Cependant bien que les critères d’hypoxémie soient présents, ce SDRA diffère des formes classiques notamment en raison d’une compliance pulmonaire le plus souvent normale au stade initial. Ceci suggère des mécanismes physiopathologiques spécifiques encore mal compris, qui aboutissent à des profils de la maladie devant faire repenser la ventilation protectrice, afin de limiter la genèse des lésions pulmonaires induites par la ventilation. Summary SARS-coV2 infection may induce a severe pneumonia that may lead to an acute respiratory distress syndrome. Hypoxaemia is the key symptom of the disease but other features are different such as pulmonary compliance that is most of the time initially normal. The mechanisms of the pulmonary damage are not completely understood. A new ventilation strategy has been set up to prevent ventilator induced lung injury (VILI).


title: Capability, opportunity, and motivation to enact hygienic practices in the early stages of the COVID‐19 outbreak in the United Kingdom

text: OBJECTIVES: The COVID‐19 pandemic is one of the greatest global health threats facing humanity in recent memory. This study aimed to explore influences on hygienic practices, a set of key transmission behaviours, in relation to the Capability, Opportunity, Motivation‐Behaviour (COM‐B) model of behaviour change (Michie et al., 2011). DESIGN: Data from the first wave of a longitudinal survey study were used, launched in the early stages of the UK COVID‐19 pandemic. METHODS: Participants were 2025 adults aged 18 and older, representative of the UK population, recruited by a survey company from a panel of research participants. Participants self‐reported motivation, capability, and opportunity to enact hygienic practices during the COVID‐19 outbreak. RESULTS: Using regression models, we found that all three COM‐B components significantly predicted good hygienic practices, with motivation having the greatest influence on behaviour. Breaking this down further, the subscales psychological capability, social opportunity, and reflective motivation positively influenced behaviour. Reflective motivation was largely driving behaviour, with those highest in reflective motivation scoring 51% more on the measure of hygienic practices compared with those with the lowest scores. CONCLUSIONS: Our findings have clear implications for the design of behaviour change interventions to promote hygienic practices. Interventions should focus on increasing and maintaining motivation to act and include elements that promote and maintain social support and knowledge of COVID‐19 transmission. Groups in particular need of targeting for interventions to increase hygienic practices are males and those living in cities and suburbs.


title: Clinical implications of rapid ePlex® Respiratory Pathogen Panel testing compared to laboratory-developed real-time PCR

text: Rapid diagnosis of respiratory infections is of great importance for adequate isolation and treatment. Due to the batch-wise testing, laboratory-developed real-time polymerase chain reaction (PCR) assays (LDT) often result in a time to result of one day. Here, LDT was compared with rapid ePlex® Respiratory Pathogen (RP) Panel testing of GenMark Diagnostics (Carlsbad, CA, USA) with regard to time to result, installed isolation precautions, and antibacterial/antiviral treatment. Between January and March 2017, 68 specimens of 64 patients suspected of an acute respiratory infection were tested with LDT and the ePlex® RP panel. The time to result was calculated as the time between sample reception and result reporting. Information regarding isolation and antibacterial/antiviral treatment was obtained from the patient records. Thirty specimens tested LDT positive (47%) and 29 ePlex® RP panel positive (45%). The median time to result was 27.1 h (range 6.5–96.6) for LDT versus 3.4 h (range 1.5–23.6) for the RP panel, p-value < 0.001. In 14 out of 30 patients, isolation was discontinued based on the ePlex® RP panel results, saving 21 isolation days. ePlex® RP panel test results were available approximately one day ahead of the LDT results in the 19 patients receiving antiviral/antibacterial treatment. In addition, two bacterial pathogens, not requested by the physician, were detected using the RP panel. Analysis of respiratory infections with the ePlex® RP panel resulted in a significant decrease in time to result, enabling a reduction in isolation days in half of the patients. Furthermore, syndromic RP panel testing increased the identification of causative pathogens.


COVID-19 Pandemic: the long and difficult way back to work


title: Viral-Mediated mRNA Degradation for Pathogenesis

text: Cellular RNA decay machinery plays a vital role in regulating gene expression by altering the stability of mRNAs in response to external stresses, including viral infection. In the primary infection, viruses often conquer the host cell’s antiviral immune response by controlling the inherently cellular mRNA degradation machinery to facilitate viral gene expression and establish a successful infection. This review summarizes the current knowledge about the diverse strategies of viral-mediated regulatory RNA shutoff for pathogenesis, and particularly sheds a light on the mechanisms that viruses evolve to elude immune surveillance during infection.


title: eIF3f: A central regulator of the antagonism atrophy/hypertrophy in skeletal muscle

text: Abstract The eukaryotic initiation factor 3 subunit f (eIF3f) is one of the 13 subunits of the translation initiation factor complex eIF3 required for several steps in the initiation of mRNA translation. In skeletal muscle, recent studies have demonstrated that eIF3f plays a central role in skeletal muscle size maintenance. Accordingly, eIF3f overexpression results in hypertrophy through modulation of protein synthesis via the mTORC1 pathway. Importantly, eIF3f was described as a target of the E3 ubiquitin ligase MAFbx/atrogin-1 for proteasome-mediated breakdown under atrophic conditions. The biological importance of the MAFbx/atrogin-1-dependent targeting of eFI3f is highlighted by the finding that expression of an eIF3f mutant insensitive to MAFbx/atrogin-1 polyubiquitination is associated with enhanced protection against starvation-induced muscle atrophy. A better understanding of the precise role of this subunit should lead to the development of new therapeutic approaches to prevent or limit muscle wasting that prevails in numerous physiological and pathological states such as immobilization, aging, denervated conditions, neuromuscular diseases, AIDS, cancer, diabetes. This article is part of a Directed Issue entitled: Molecular basis of muscle wasting.


title: Infectious Complications After Umbilical Cord Blood Transplantation for Hematological Malignancy

text: BACKGROUND: Umbilical cord blood transplant (UCBT) is used for patients who do not have a matched donor, but engraftment often takes longer than with a standard allogeneic transplant, likely increasing the risk for infection. We characterized specific infections and outcomes in adults undergoing UCBT at our 2 centers. METHODS: All adults who underwent UCBT between January 1, 2006 and December 31, 2015 were included. Infectious episodes from 6 months before to 2 years after UCBT were reviewed. RESULTS: Fifty-seven patients underwent UCBT; 47 had neutrophil engraftment. A total of 179 infectious episodes occurred in 55 patients, 73 (41%) within 30 days post-UCBT. Viruses caused 85 (47%) infections. Cytomegalovirus caused 32 infectious episodes and was most common from day 30 to 100. Human herpesvirus 6 occurred in 28 episodes, was most common within 30 days, and caused 1 death. Bacteria were responsible for 82 (46%) infections, most commonly bacteremias due to Staphylococcus spp, Enterococcus spp, and Enterobacteriaceae. Of 11 invasive fungal infections, 9 were aspergillosis, 4 of which were fatal. Overall mortality was 56% in the first year. Thirteen deaths were from infection; 11 occurred in the first 100 days and 7 in the first 30 days post-UCBT. Of 10 patients who never engrafted, 9 died, 6 from infection, within 100 days post-UCBT. CONCLUSIONS: Infectious complications were common after UCBT, especially in the first 30 days. Deaths from viral infections were fewer than expected. Delayed engraftment and nonengraftment continue to convey increased risk for fatal bacterial and fungal infections post-UCBT.


title: Community-Acquired Pneumonia Hospitalization among Children with Neurologic Disorders

text: Objective To describe and compare the clinical characteristics, outcomes, and etiology of pneumonia among children hospitalized with community-acquired pneumonia (CAP) with neurologic disorders, non-neurologic underlying conditions, and no underlying conditions. Study design Children <18 years old hospitalized with clinical and radiographic CAP were enrolled at 3 US children's hospitals. Neurologic disorders included cerebral palsy, developmental delay, Down syndrome, epilepsy, non-Down syndrome chromosomal abnormalities, and spinal cord abnormalities. We compared the epidemiology, etiology, and clinical outcomes of CAP in children with neurologic disorders with those with non-neurologic underlying conditions, and those with no underlying conditions using bivariate, age-stratified, and multivariate logistic regression analyses. Results From January 2010-June 2012, 2358 children with radiographically confirmed CAP were enrolled; 280 (11.9%) had a neurologic disorder (52.1% of these individuals also had non-neurologic underlying conditions), 934 (39.6%) had non-neurologic underlying conditions only, and 1144 (48.5%) had no underlying conditions. Children with neurologic disorders were older and more likely to require intensive care unit (ICU) admission than children with non-neurologic underlying conditions and children with no underlying conditions; similar proportions were mechanically ventilated. In age-stratified analysis, children with neurologic disorders were less likely to have a pathogen detected than children with non-neurologic underlying conditions. In multivariate analysis, having a neurologic disorder was associated with ICU admission for children ≥2 years of age. Conclusions Children with neurologic disorders hospitalized with CAP were less likely to have a pathogen detected and more likely to be admitted to the ICU than children without neurologic disorders.


title: What needs to be done to control the spread of Middle East respiratory syndrome coronavirus?

text: Up to November 2014, Middle East respiratory syndrome coronavirus (MERS-CoV) has infected 935 individuals and killed 371, all originating in or with links to the Middle East. The mechanisms of transmission of the disease are not fully understood, but MERS-CoV seems to sustain itself in the human population through repeated re-introduction from a camel reservoir and is able to cause nosocomial outbreaks. The risk of a global spread of MERS-CoV is low. Epidemiological, serological and phylogenetic research, combined with one health surveillance, dynamic case definitions, active case finding, rigorous infection control, culturally sensitive risk communication and a continuous re-evaluation of new evidence will enable to better understand the disease, limit its spread and quantify its risk in order to better prepare for a hypothetical spread.


title: Super induction of dengue virus NS1 protein in E. coli

text: Abstract The non-structural protein 1 (NS1) of dengue virus is a useful target for diagnostics of dengue infection since the protein is abundantly circulating in blood during the acute phase of the disease. Prior work has established that secreted NS1 levels in plasma correlates with viremia levels and hence can also be used to diagnose patients at the risk for developing dengue hemorrhagic fever. Thus detection of non-structural dengue antigens may be of benefit for an early rapid diagnosis of dengue infection due to its long half life in the blood. Here we describe a simple and efficient method for the expression of NS1 in Escherichia coli, which could potentially be used to develop monoclonal and bispecific antibodies for point of care diagnostics. E. coli codon optimized synthetic full-length NS1 gene of dengue serotype 1 (DEN-1) was successfully cloned and expressed in very high-level as inclusion bodies. The NS1 protein was successfully affinity purified and refolded as a recombinant NS1 (rNS1) protein in E. coli and yield was 230–250mg/L of bacterial culture. The rNS1 protein was used to immunize mice for hybridoma development. The polyclonal antiserum from animals immunized with this rNS1 protein was found to specifically recognize the rNS1, thus demonstrating the immunogenic nature of the protein. The rNS1 protein purified from E. coli could be useful for developing a sensitive serum diagnostic assay to monitor dengue outbreaks.


title: Evaluation of the immune response in conventionally weaned pigs infected with porcine deltacoronavirus

text: Although porcine deltacoronavirus (PDCoV) is a significant pandemic threat in the swine population and has caused significant economic losses, information regarding the immune response in conventionally weaned pigs infected with PDCoV is scarce. Hence, the immune response in conventionally weaned pigs infected with PDCoV was assessed after challenge and rechallenge. After the first challenge, obvious diarrhea and viral shedding developed successively in all pigs in the four inoculation dose groups from 3 to 14 days postinfection (dpi), and all pigs recovered (no clinical symptoms or viral shedding) by 21 dpi. All pigs in the four groups exhibited significantly increased PDCoV-specific IgG, IgA and virus-neutralizing (VN) antibody (Ab) titers and IFN-γ levels in the serum after the first challenge. All pigs were completely protected against rechallenge at 21 dpi. The serum levels of PDCoV-specific IgG, IgA, and VN Abs increased further after rechallenge. Notably, the IFN-γ level declined continuously after 7 dpi. In addition, the levels of PDCoV-specific IgG, IgA and VN Abs in saliva increased significantly after rechallenge and correlated well with the serum Ab titers. Furthermore, the appearance of clinical symptoms of PDCoV infection in conventionally weaned pigs was delayed with reduced inoculation doses. In summary, the data presented here offer important reference information for future PDCoV animal infection and vaccine-induced immunoprotection experiments.


title: Detection of influenza C viruses among outpatients and patients hospitalized for severe acute respiratory infection, Minnesota, 2013–2016

text: BACKGROUND: Existing literature suggests that influenza C typically causes mild respiratory tract disease. However, clinical and epidemiological data are limited. METHODS: Four outpatient clinics and three hospitals submitted clinical data and respiratory specimens through a surveillance network for acute respiratory infection (ARI) during May 2013 through December 2016. Specimens were tested using multi-target nucleic acid amplification tests (NAAT) for 19–22 respiratory pathogens, including influenza C. RESULTS: Influenza C virus was detected among 59 of 10,202 (0.58%) hospitalized SARI cases and 11 of 2,282 (0.48%) outpatients. Most detections occurred from December to March, with 73% during the 2014–2015 season. Influenza C detections occurred among patients of all ages, with similar rates between inpatients and outpatients. The highest rate of detection occurred among children aged 6 to 24 months (1.2%). Among hospitalized cases, seven required intensive care. Medical co-morbidities were reported in 58% of hospitalized cases and all who required intensive care. At least one other respiratory pathogen was detected in 40 (66%) cases, most commonly rhinovirus/enterovirus (25%) and respiratory syncytial virus (RSV) (20%). The hemagglutinin-esterase-fusion (HEF) gene was sequenced in 37 specimens, and both C/Kanagawa and C/Sao Paulo lineages were detected in inpatients and outpatients. CONCLUSIONS: We found seasonal circulation of influenza C with year-to-year variability. Detection was most frequent among young children, but occurred in all ages. Some cases positive for influenza C, particularly those with co-morbid conditions, had severe disease, suggesting a need for further study of the role of influenza C virus in the pathogenesis of respiratory disease.


title: Detection of alpha- and betacoronaviruses in rodents from Yunnan, China

text: BACKGROUND: Rodents represent the most diverse mammals on the planet and are important reservoirs of human pathogens. Coronaviruses infect various animals, but to date, relatively few coronaviruses have been identified in rodents worldwide. The evolution and ecology of coronaviruses in rodent have not been fully investigated. RESULTS: In this study, we collected 177 intestinal samples from thress species of rodents in Jianchuan County, Yunnan Province, China. Alphacoronavirus and betacoronavirus were detected in 23 rodent samples from three species, namely Apodemus chevrieri (21/98), Eothenomys fidelis (1/62), and Apodemus ilex (1/17). We further characterized the full-length genome of an alphacoronavirus from the A. chevrieri rat and named it as AcCoV-JC34. The AcCoV-JC34 genome was 27,649 nucleotides long and showed a structure similar to the HKU2 bat coronavirus. Comparing the normal transcription regulatory sequence (TRS), 3 variant TRS sequences upstream the spike (S), ORF3, and ORF8 genes were found in the genome of AcCoV-JC34. In the conserved replicase domains, AcCoV-JC34 was most closely related to Rattus norvegicus coronavirus LNRV but diverged from other alphacoronaviruses, indicating that AcCoV-JC34 and LNRV may represent a novel alphacoronavirus species. However, the S and nucleocapsid proteins showed low similarity to those of LRNV, with 66.5 and 77.4% identities, respectively. Phylogenetic analysis revealed that the S genes of AcCoV-JC34, LRNV, and HKU2 formed a distinct lineage with all known coronaviruses. CONCLUSIONS: Both alphacoronaviruses and betacoronaviruses were detected in Apodemus chevrieri in the Yunnan Province of China, indicating that Apodemus chevrieri is an important host for coronavirus. Several new features were identified in the genome of an Apodemus chevrieri coronavirus. The phylogenetic distance to other coronaviruses suggests a variable origin and evolutionary route of the S genes of AcCoV-JC34, LRNV, and HKU2. These results indicate that the diversity of rodent coronaviruses is much higher than previously expected. Further surveillance and functional studies of these coronaviruses will help to better understand the importance of rodent as host for coronaviruses. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12985-017-0766-9) contains supplementary material, which is available to authorized users.


title: Occupational infection and needle stick injury among clinical laboratory workers in Al-Madinah city, Saudi Arabia

text: BACKGROUND: Clinical laboratory workers face biohazard such as needlestick injury and occupational infection on a daily basis. In this study, we examined self-reported frequency of occupational infection and needlestick injury among the clinical laboratory workers in Al- Madinah, Saudi Arabia. METHODS: A total of 234 clinical laboratory workers were recruited from private and government health sectors to answer a self-administered questionnaire that was prepared to achieve the aims of the study. RESULTS: The results showed that approximately 33% of the sample had an experienced occupational infection while 24% had experienced a needlestick injury. Approximately, 49% reported that they always recap needle after use, whereas 15% reported doing that most of the times. Occupational infection, needlestick injury and recapping needles after use were associated with lack of training on biosafety (P < 0.05). CONCLUSION: The frequency of occupational infection and needlestick injury among clinical laboratory workers in Al-Madinah is high. Interventions related to biosafety and infection control and the use of needlestick prevention devices might be useful in lowering such frequency.


title: Vascular Targeting: Recent Advances and Therapeutic Perspectives

text: The ability to deliver therapeutics site—specifically in vivo—remains a major challenge for the treatment of malignant, inflammatory, cardiovascular, and degenerative diseases. The need to target agents safely, efficiently, and selectively has become increasingly evident because of progress in vascular targeting. The vascular endothelium is a central target for intervention, given its multiple roles in the physiology (in health) and pathophysiology (in disease) and its direct accessibility to circulating ligands. In cancer, the expression of specific molecules on the surface of vascular endothelial and perivascular cells might enable direct therapeutic targeting. The use of in vivo phage display has significantly contributed to the identification of such targets, which have been successfully used for directed vascular targeting in preclinical animal models. Several animal studies have been performed by using fused molecules between tumor endothelium-directed molecules and immunomodulatory, procoagulant, or cytotoxic molecules. In addition to delivery of therapeutic agents, vascular targeted gene therapies based on both ligand-directed delivery of gene vectors to tumor endothelium and transcriptional targeting have also emerged. In this review, we discuss ligand-directed vascular targeting strategies with an emphasis on recent developments related to phage-display-based screenings.


title: Policy Decisions and Use of Information Technology to Fight Coronavirus Disease, Taiwan

text: Because of its proximity to and frequent travelers to and from China, Taiwan faces complex challenges in preventing coronavirus disease (COVID-19). As soon as China reported the unidentified outbreak to the World Health Organization on December 31, 2019, Taiwan assembled a taskforce and began health checks onboard flights from Wuhan. Taiwan’s rapid implementation of disease prevention measures helped detect and isolate the country’s first COVID-19 case on January 20, 2020. Laboratories in Taiwan developed 4-hour test kits and isolated 2 strains of the coronavirus before February. Taiwan effectively delayed and contained community transmission by leveraging experience from the 2003 severe acute respiratory syndrome outbreak, prevalent public awareness, a robust public health network, support from healthcare industries, cross-departmental collaborations, and advanced information technology capacity. We analyze use of the National Health Insurance database and critical policy decisions made by Taiwan’s government during the first 50 days of the COVID-19 outbreak.


title: SARS-CoV-2 and Legionella Co-infection in a Person Returning from a Nile Cruise

text: We report a case of severe acute respiratory syndrome coronavirus 2 and Legionella co-infection manifesting as pneumonia with gastrointestinal symptoms. The case highlights the importance of differential diagnosis during the COVID-19 pandemic, so we do not miss the opportunity to diagnose other treatable causes of disease with similar symptoms.


title: GP73, a novel Golgi-localized protein upregulated by viral infection

text: Abstract We report the isolation and characterization of GP73, a novel 73kDa human Golgi protein. The GP73 cDNA was cloned by differential screening of a cDNA library derived from the liver of a patient with adult giant-cell hepatitis (GCH), a rare form of hepatitis with presumed viral etiology. In vitro transcription–translation studies indicate that GP73 is an integral membrane protein, and immunolocalization experiments using epitope-tagged GP73 demonstrate that the protein is localized to the Golgi apparatus. Northern blot analysis of RNA from multiple human tissues reveals a single GP73 mRNA transcript with a size of approximately 3.0kb. Immunohistochemical studies using rabbit polyclonal antisera directed against recombinant GP73 demonstrate that the protein is preferentially expressed by epithelial cells in many human tissues. In normal livers, GP73 is consistently present in biliary epithelial cells, whereas hepatocytes show little or no signal. In contrast, livers of patients with GCH display strong GP73 immunoreactivity in multinucleated hepatocytes. GP73 mRNA and protein are expressed in highly differentiated HepG2 hepatoma cells after infection with adenovirus in vitro. We conclude that GP73 represents a novel, epithelial cell-specific integral membrane Golgi protein that can be upregulated in response to viral infection.


title: Etiología de bronquiolitis en niños hospitalizados en el sureste de España

text: Resumen Introducción La bronquiolitis es una de las infecciones respiratorias más comunes en niños menores de 2 años y una de las principales causas de ingreso en niños pequeños, especialmente durante el invierno. Objetivos Determinar la prevalencia y etiología de bronquiolitis en el sureste de España. Material y métodos Se realizó un estudio prospectivo durante la temporada de bronquiolitis (diciembre-abril). En el estudio se incluyeron niños menores de 18 meses ingresados en el hospital con motivo del primer episodio de bronquiolitis. Se analizaron aspirados nasofaríngeos mediante una técnica de reacción en cadena de la polimerasa (múltiple), precedida de transcripción inversa del ARN viral (RT-PCR). Resultados Se incluyeron en el estudio 235 niños, de cada uno de los cuales se obtuvo una muestra de aspirado nasofaríngeo. Se detectaron virus respiratorios en muestras de 204 niños (en total 287 virus). El virus respiratorio sincitial se detectó con mayor frecuencia seguido del rinovirus. Se observaron infecciones virales mixtas en el 36% de los niños en los que se detectó algún virus respiratorio. La asociación más frecuente fue el virus respiratorio sincitial A con rinovirus. Conclusiones Se detectaron virus respiratorios en la mayoría de los niños menores de 18 meses hospitalizados por bronquiolitis, en forma de infección mixta en el 36% de aquellos con infección viral. Abstract Background Bronchiolitis is the most common respiratory disease in children under 2 years-old and a major cause of hospitalization in young children, especially during the winter. Objectives To determine the prevalence and etiology of bronchiolitis in south-east of Spain. Study design A prospective study was conducted during the bronchiolitis season (December-April). Children below 18 months-old admitted to the hospital for a first bronchiolitis episode were included. Nasopharyngeal aspirates were analysed by reverse transcription polymerase chain reaction (RT-PCR) respiratory syncytial virus. Results A total of 235 children were included during this period, and 235 RT-PCR were performed. A total of 287 viruses were detected in nasopharyngeal aspirates from 204 infants. Respiratory syncytial virus was the virus detected more frequently, followed by rhinovirus. Co-infections were found in the 36% of children. Conclusions Respiratory viruses were detected in most of the children below 18 months-old hospitalised with bronchiolitis, and 36% of them showed a mixed infection.


title: Hypoxia-like tissue injury as a component of multiple sclerosis lesions

text: Abstract Recent data suggest that the mechanisms of demyelination and tissue damage in multiple sclerosis (MS) are heterogenous. In this review, evidence is discussed, which show that in a subset of multiple sclerosis patients the central nervous system (CNS) lesions show profound similarities to tissue alterations found in acute white matter stroke, thus suggesting that a hypoxia-like metabolic injury is a pathogenetic component in a subset of inflammatory brain lesions. Both, vascular pathology as well as metabolic disturbances induced by toxins of activated macrophages and microglia may be responsible for such lesions in multiple sclerosis.


COVID‐19 policy measures—Advocating for the inclusion of the social determinants of health in modelling and decision making


title: Detection of Human Metapneumovirus in Hospitalized Children with Acute Respiratory Tract Infection Using Real-time RT-PCR in a Hospital in Northern Taiwan

text: Background/purpose Human metapneumovirus (hMPV) is a newly discovered respiratory pathogen. This prospective hospital-based study investigated the clinical role and features of hMPV in Taiwan. Methods Respiratory specimens collected from hospitalized children with acute respiratory tract infection between September 1, 2003 and April 10, 2005 were screened for metapneumovirus using real-time reverse transcription-polymerase chain reaction (RT-PCR). Results During the study period, 930 specimens were obtained from 926 hospitalized children. After exclusion of 200 cases due to lack of clinical evidence of airway infection or diseases with known etiology, 726 were included in the analysis. Among these, 33 children had a positive result for hMPV infection. The majority of these patients were admitted during spring and early summer. Twenty-one (63.6%) were younger than 2 years of age. hMPV accounted for 13.3% of respiratory infections occurring between the ages of 18 and 24 months and was as common a respiratory pathogen as respiratory syncytial virus (RSV) in that age group. The 11 patients (33.3%) with underlying diseases had a similar disease course to those without underlying diseases. A co-pathogen was found in 11 patients (33.3%). Infected children between 2 and 5 years of age had significantly higher titers of hMPV in their respiratory specimens (103.88 copies/mL) than children younger than 2 years (102.26 copies/mL) (p = 0.013) and children older than 5 years (102.25 copies/mL) (p = 0.005). hMPV positive cases were significantly older than those with RSV infection (p = 0.002) and had a shorter duration of hospitalization (p = 0.001), fewer days of oxygen use (p = 0.001) and higher levels of C-reactive protein (p = 0.004). Conclusion Metapneumovirus circulates in children in northern Taiwan during spring and early summer. hMPV was the most common respiratory pathogen in children aged between 18 and 24 months hospitalized with acute respiratory tract infection. Real-time RT-PCR is a sensitive method for investigating the epidemiology and diseases associated with hMPV. [J Formos Med Assoc 2007;106(1):16-24]


Distance learning and telemedicine in the area of Otorhinolaryngology: lessons in times of pandemic


title: Emerging viral diseases and infectious disease risks

text: Summary. New pathogens and antimicrobial‐resistant forms of older pathogens continue to emerge, some with the potential for rapid, global spread and high morbidity and mortality. Pathogens can emerge either through introduction into a new population or when the interaction with the vector changes; emergence is also influenced by microbiological adaptation and change, global travel patterns, domestic and wild animal contact and other variants in human ecology and behaviour. Quick, decisive action to detect and control novel pathogens, and thereby contain outbreaks and prevent further transmission, is frequently hampered by incomplete or inadequate data about a new or re‐emerging pathogen. Three examples of pathogens that are current causes for human health concern are avian influenza, West Nile virus (WNV) and the severe acute respiratory syndrome (SARS) coronavirus. Pathogens directly or indirectly transmitted by aerosolized droplets, such as avian influenza and SARS, pose considerable containment challenges. Rapid screening tests for other newly described pathogens such as WNV require time for development and may be <100% reliable. The importance of vigilance in the detection and control of newly recognized infectious threats cannot be overstressed. The presence of infectious agents in the blood supply could again have a significant impact on the safe use of both blood and blood‐derived products in the care of patients with haemophilia, as did the human immunodeficiency virus in the 1980s. Emerging pathogens will continue to be a reality requiring the collaborative efforts of public health and individual healthcare providers worldwide to contain outbreaks and prevent transmission.


title: Characteristics of a family cluster of Severe Acute Respiratory Syndrome Coronavirus 2 in Henan, China

text: • The worldwide outbreak of SARS-CoV-2 called for more close investigations of the family transmission routes of infectious diseases. • We hypothesize that the likelihood to develop COVID-19 is probably associated with the duration of being exposed to SARS-CoV-2. • In a family cluster, asymptomatic infection carriers can be persons of all ages, from nine to sixty years old.


title: The Frankfurt COVid aErosol pRotEction Dome – COVERED- a consideration for personal protective equipment improvement and technical note

text: Abstract In times of global pandemics, the self-protection of medical personnel is of paramount importance. Faced with the worldwide supply shortages of medical equipment, we have designed a simple, effective splash protection made of medical packaging. Because of the ease of manufacture and the instructions published in this technical note, we hope that healthcare professionals worldwide will be able to protect themselves against infectious fluids with an intubation dome.


title: Duration of Antibody Responses after Severe Acute Respiratory Syndrome

text: Among 176 patients who had had severe acute respiratory syndrome (SARS), SARS-specific antibodies were maintained for an average of 2 years, and significant reduction of immunoglobulin G–positive percentage and titers occurred in the third year. Thus, SARS patients might be susceptible to reinfection >3 years after initial exposure.


title: Increase in placental apolipoprotein D as an adaptation to human gestational diabetes

text: Abstract The expression of apolipoprotein D (apo D), a lipocalin involved in defense mechanisms against oxidative stress, in placental tissue samples of pregnancies with gestational diabetes mellitus (GDM) was compared to non-diabetic controls. We have investigated the relationship of apo D with 4-HNE, a major propagation product of lipid peroxidation, in stressed tissues. We included 20 pregnant women with GDM and 30 women with normal ongoing pregnancies as the control group. Placentas were collected and frozen for Western blot or included in paraffin for immunohistochemistry. The intensity of immunostaining was higher for apo D and 4-HNE in GDM samples; however, the differences in expression between the groups was more intense for apo D. Positive signals for both antibodies was detected in the villous trophoblast and adventitia tunica around the large blood vessels for all groups. Specific immunostaining for apo D was noted in some mesenchymal and macrophagic-like cells and this signal increased in diabetic placentas. Densitometry analysis of Western blots showed no significant difference for 4-HNE, but was significantly more intense for apo D in diabetic women. The contradictory results for 4-HNE could be due to changes which are too small and are masked in tissue homogenates. The results for apo D showed a strong relationship with GDM in the placenta that may reflect its suggested function in defense mechanisms against oxidative stress.


title: Making difficult ethical decisions in patient care during natural disasters and other mass casualty events

text: Objective Recent experiences in the United States with unprecedented terrorist attacks (9/11) and a devastating natural disaster (Hurricane Katrina) have demonstrated that the medical care of mass casualties during such disasters poses ethical problems not normally experienced in civilian health care. It is important to 1) identify the unique ethical challenges facing physicians who feel an obligation to care for victims of such disasters and 2) develop a national consensus on ethical guidelines as a resource for ethical decision making in medical disaster relief. Study Design A survey of pertinent literature was performed to assess experience and opinions on the condition of medical care in terrorist attacks and natural disasters, the ethical challenges of disaster medical care, and the professional responsibilities and responsiveness in disasters. Conclusions It is necessary to develop a national consensus on the ethical guidelines for physicians who care for patients, victims, and casualties of disasters, and to formulate a virtue-based, yet practical, ethical approach to medical care under such extreme conditions. An educational curriculum for medical students, residents, and practicing physicians is required to best prepare all physicians who might be called upon, in the future, to triage patients, allocate resources, and make difficult decisions about treatment priorities and comfort care. It is not appropriate to address these questions at the time of the disaster, but rather in advance, as part of the ethics education of the medical profession. Important issues for resolution include inpatient and casualty triage and prioritization, medical liability, altered standards of care, justice and equity, informed consent and patient autonomy, expanding scope of practice in disaster medicine, and the moral and ethical responsibilities of physicians to care for disaster victims.


title: Literature-related discovery and innovation — update

text: Abstract Literature-Related Discovery and Innovation (LRDI — formerly LRD — literature-related discovery) integrates 1) discovery generation from disparate literatures with 2) the wealth of knowledge contained in prior art to 3) potentially reverse chronic and infectious diseases and/or 4) potentially solve technical problems that appear intractable. This article describes the evolution of LRDI by the author and the insights gained/lessons learned over the past decade. To illustrate the potential power of LRDI, the article emphasizes the relationship between the results of our 2008 LRDI multiple sclerosis (MS) study and a recent demonstration of MS reversal. Lessons learned from the six LRDI medical studies done so far include: ⁎ The main operational problem in the author's LRDI approach is selecting the most important concepts from extremely large volumes of potential discovery retrieval. This is contrary to most published LRDI research, where the discovery focus is searching for rare events. ⁎ It is important to have topical specialist(s) working closely with information technologist(s); the topical specialist(s) applies judgment in selecting the most important concepts. ⁎ A functional form of the information retrieval query with proximity searching capability provides highly selective filtering for discovery retrieval and core prevention/treatment retrieval; the functional form of the query with proximity searching capability allows the use of full-text for discovery and core prevention/treatment. ⁎ Bibliographic coupling (identifying papers that share common references) combined with text-based relationships strengthens selection for potential discovery further. ⁎ Having ‘skin-in-the-game’ (being affected personally) relative to the medical outcome is a strong incentive to do whatever is necessary to solve the research problem. ⁎ Hormesis is critical to healing; relatively modest doses of stimuli tend to be beneficial, whereas relatively large doses may be harmful. The synergy of hormetic treatment doses produces effects larger than combinations of individual doses and requires smaller doses when combined; the synergy of hormetic doses allows conversion of megadoses of nutrients typically reported in lab/clinical studies to physiological (food-level) doses and associated increased safety. ⁎ Co-promoters (combinations of toxic stimuli required to produce disease symptoms) are extremely important for explaining seemingly conflicting results; if true co-promotion is present, elimination of one of the co-promoters may be adequate for removing symptoms, even though the overall problem persists. ⁎ Prior art (potential treatments already published in the literature but not pursued by mainline medicine) may have much to contribute to potentially solve many serious medical problems; much of prior art is overlooked, especially low-tech prior art (e.g., foods, food extracts, herbs, etc.). ⁎ Systemic and focused treatments are both necessary components of healing, but neither will be fully, or many times even partially, effective until the cause(s) is identified and removed. Any medical approach that involves administering treatments for chronic and infectious diseases without addressing the cause(s) results in a broad range of outcomes mainly involving substitution of one set of symptoms for another. ⁎ Past results of LRDI medical studies showed much overlap among preventatives/systemic treatments for different diseases. Differences will arise mainly in focused treatments, especially those involving high technology. ⁎ The central parameters to healing in much medical research are never identified nor reported. Many treatments require a combination of skilled practitioners, cause removal, and immune/neural/endocrine/circulatory systems to be healthy for full effectiveness, yet practitioner skill, degree of cause removal, and immune system et al. health are never reported. A lack of this information does not allow efficacy of different treatments to be compared. Reviews and meta-analyses that compare and draw conclusions about the effectiveness of these different treatments without the above critical information being reported are of extremely limited value and credibility. ⁎ Finally, the most important deficiency for fully reversing chronic and infectious diseases, as well as rapidly accelerating healing of injuries and wounds, is the credibility and integrity of the medical literature itself, especially in areas that concern commercial and government/political sensitivities. In the evaluation of many concepts that deviated from the norm, it was difficult to ascertain whether the difference was based on solid high-quality research, poor research, or deliberately skewed research.


title: Targeting the viral Achilles’ heel: recognition of 5′-triphosphate RNA in innate anti-viral defence

text: Some RNA virus genomes bear 5′-triphosphates, which can be recognized in the cytoplasm of infected cells by host proteins that mediate anti-viral immunity. Both the innate sensor RIG-I and the interferon-induced IFIT proteins bind to 5′-triphosphate viral RNAs. RIG-I signals for induction of interferons during RNA virus infection while IFITs sequester viral RNAs to exert an anti-viral effect. Notably, the structures of these proteins reveal both similarities and differences, which are suggestive of independent evolution towards ligand binding. 5′-triphosphates, which are absent from most RNAs in the cytosol of uninfected cells, are thus a marker of virus infection that is targeted by the innate immune system for both induction and execution of the anti-viral response.


title: Development and evaluation of a novel bioaerosol amplification unit (BAU) for improved viral aerosol collection

text: Abstract A novel bioaerosol amplification unit (BAU) that increases the size of viral particles by condensational growth has been designed and evaluated for improved viral aerosol collection. In the BAU, water was used as the condensing vapor to preserve viability of virus, and supersaturation conditions for condensational growth of particles were achieved by either conductive cooling or mixing with hot, water-saturated air. MS2 bacteriophage (28nm) was used as the test agent, and changes in collection efficiency of an SKC Biosampler with and without the BAU were determined by assaying plaque-forming units (PFUs) in the collection medium. Results showed that the mixing-type BAU (mBAU) was a promising device for improved viral aerosol sampling. The number of viruses (PFU) collected in the Biosampler increased 2–3 fold after passing through the mBAU. However, PFU increases in the cooling-type BAU (cBAU) were insignificant. APS results likewise showed that the mBAU was better in growing particles than the cBAU. After growth, number concentrations of particles larger than 327nm in the cBAU and mBAU increased 1.3 and 15.0 fold, respectively. The relatively high molecular diffusivity of water vapor compared to the thermal diffusivity of air and the temperature gradient in the cBAU tube limited particle growth by causing condensation to occur predominantly at the colder wall.


title: Ultrastructure of human nasal epithelium during an episode of coronavirus infection

text: The nasal epithelium from a young girl was examined by electron microscopy and found to be infected by coronavirus. Virions are seen within and outside the ciliated cells, but not outside or within the goblet cells or other cells of the nasal mucosa. Some virions are located near the microvilli, others in pockets in the apical cell membrane. The cytoplasm contains many small vesicles with a single virion, large apical vesicles containing hundreds of virions, and lysosome-like cytosomes with a moderate number of virions. Some viruslike particles devoid of an electron-dense interior are seen both in the cytosomes and extracellularly. Virus budding was observed in the Golgi apparatus but nowhere else in the cell. The ciliated cells seem not to be destroyed by the viruses, although in many cases the cilia are withdrawn into the cell body. The loss of cilia is likely to cause rhinorrhoea.


title: Glial Cell Expression of PD-L1

text: The programmed death (PD)-1/PD-L1 pathway is a well-recognized negative immune checkpoint that results in functional inhibition of T-cells. Microglia, the brain-resident immune cells are vital for pathogen detection and initiation of neuroimmune responses. Moreover, microglial cells and astrocytes govern the activity of brain-infiltrating antiviral T-cells through upregulation of PD-L1 expression. While T-cell suppressive responses within brain are undoubtedly beneficial to the host, preventing cytotoxic damage to this vital organ, establishment of a prolonged anti-inflammatory milieu may simultaneously lead to deficiencies in viral clearance. An immune checkpoint blockade targeting the PD-1: PD-L1 (B7-H1; CD274) axis has revolutionized contemporary treatment for a variety of cancers. However, the therapeutic potential of PD1: PD-L1 blockade therapies targeting viral brain reservoirs remains to be determined. For these reasons, it is key to understand both the detrimental and protective functions of this signaling pathway within the brain. This review highlights how glial cells use PD-L1 expression to modulate T-cell effector function and limit detrimental bystander damage, while still retaining an effective defense of the brain.


title: Evaluation of the chronic disease self-management program in a Chinese population

text: Abstract Objective This study evaluated the 6-week Chronic Disease Self-Management Program (CDSMP) in Hong Kong. Methods A total of 148 subjects with chronic illness were recruited. Subjects were matched on duration of illness and gender, and then randomly allocated to experimental and comparison groups. The experimental group participated in the CDSMP, while the comparison group joined a Tai-Chi interest class in a mass-activity format. Subjects completed evaluation questionnaires before beginning their program and 1 week following the program. Results Analysis of covariance showed that the CDSMP participants demonstrated significantly higher self-efficacy in managing their illness, used more cognitive methods to manage pain and symptoms, and felt more energetic than the subjects in the comparison group. The CDSMP participants also demonstrated changes in their profile of coping strategies, having a tendency to adopt the cognitive methods of diverting attention, reinterpreting pain, ignoring sensations, and making positive self-statements. Conclusion The short-term evaluation results showed that the CDSMP primarily increased the self-efficacy, exercise behavior, and application of cognitive coping strategies of the participants. Practice Implication The effect of the CDSMP in a Chinese population is similar to that found in studies in Western cultures, and the CDSMP could be applied effectively in a Chinese population.


title: Chapter 6. Protection of patients and staff during a pandemic

text: PURPOSE: To provide recommendations and standard operating procedures (SOPs) for intensive care unit (ICU) and hospital preparations for an influenza pandemic or mass disaster with a specific focus on protection of patients and staff. METHODS: Based on a literature review and expert opinion, a Delphi process was used to define the essential topics including protection of patients and staff. RESULTS: Key recommendations include: (1) prepare infection control and occupational health policies for clinical risks relating to potential disease transmission; (2) decrease clinical risks and provide adequate facilities through advanced planning to maximise capacity by increasing essential equipment, drugs, supplies and encouraging staff availability; (3) create robust systems to maintain staff confidence and safety by minimising non-clinical risks and maintaining or escalating essential services; (4) prepare formal reassurance plans for legal protection; (5) provide assistance to staff working outside their normal domains. CONCLUSIONS: Judicious planning and adoption of protocols for protection of patients and staff are necessary to optimise outcomes during a pandemic.


title: Exploring milk shipment data for their potential for disease monitoring and for assessing resilience in dairy farms

text: Abstract The use of routinely recorded data for research purposes and disease surveillance is an attractive proposition. However, this requires that the validity and reliability of the data be evaluated for the purpose for which they are to be used. This manuscript reports an evaluation of milk shipment data for evaluating their usefulness in disease monitoring and the resilience of organic and conventional dairy herds in Sweden. A large number of inconsistencies were observed in the data, necessitating substantial efforts to “clean” the data. Given that the selection of rules used in the cleaning process was subjective in nature, a sensitivity analysis was carried out to determine if different cleaning routines produced substantially different results. Despite the cleaning efforts we observed far more large residuals at the shipment level than expected. Thus, it was concluded that the data were too “noisy” to be used for identification of short term impacts on milk production. Resilience was evaluated by examining the residual variance in milk shipped per cow per day under the assumption that herds with high resilience would have lower residual variance. The effects on residual variance of organic status or whether or not the herd used an automatic milking system were evaluated in models in which the residual variance was stratified or not by these factors. We did not find consistent evidence to suggest that organic herds had higher resilience than conventional herds, but this could be partly due to using residual variance as the measure indicating resilience.


title: Rotavirus y adenovirus 40–41 como causantes de gastroenteritis infantil

text: RESUMEN Introducción El rotavirus es la principal causa de diarrea grave en niños menores de 5 años, y ocasiona una elevada morbilidad y mortalidad a escala mundial. El objetivo de este trabajo ha sido conocer la epidemiología de los virus entéricos que se declaran en el Sistema de Notificación Microbiológica de Cataluña (SNMC), rotavirus y adenovirus 40-41, y valorar la importancia que tienen en el contexto de la enfermedad entérica. Material y métodos Se han revisado retrospectivamente las declaraciones que realizan los laboratorios de los hospitales que participan en el SNMC durante los años 1995-2002. Los casos de gastroenteritis aguda se han diagnosticado por detección de antígeno en heces. Las variables analizadas han sido: edad, sexo, mes y año de aislamiento. Como métodos estadísticos se han utilizado el test de la ?2 y test de Fisher; las proporciones se han comparado utilizando la prueba binomial y el nivel de significación estadística se ha establecido en 0,05. Resultados Se han detectado 6.708 (94,3%) rotavirus y 409 (5,7%) adenovirus 40-41. Las gastroenteritis agudas virales han presentado mayor morbilidad en niños pequeños menores de 2 años: el 76 y el 68% de rotavirus y adenovirus, respectivamente, se han concentrado en este grupo de edad. El rotavirus se ha aislado principalmente en invierno (68%), mientras que el adenovirus, aunque se ha presentado estable en todas las estaciones, ha sido más frecuente en otoño (35,2%). Respecto a las enteritis, bacterianas y virales, vigiladas en el SNMC, el rotavirus y el adenovirus han supuesto la segunda y la cuarta causa etiológica, respectivamente, en los niños menores de 5 años, después de Campylobacter y Salmonella. Conclusiones De los diferentes virus que pueden causar gastroenteritis aguda, hay otros virus con frecuencias elevadas, como calici-virus y astrovirus, que no se buscan sistemáticamente, por lo que hay que valorar las frecuencias con prudencia. A pesar de esto, el rotavirus es la principal causa de la gastroenteritis viral, y afecta principalmente a edades tempranas de la vida, sobre todo a niños de 1 a 11 meses, con un claro predominio en invierno. Respecto al adenovirus 40-41, se ha presentado preferentemente en niños menores de 2 años y con mayor frecuencia en otoño. La vacunación contra el rotavirus es prioritaria para poder disminuir el alto índice de morbilidad y mortalidad que ocasiona esta enfermedad. ABSTRACT Introduction Rotavirus is the main cause of severe diarrhea in children aged less than 5 years, leading to elevated morbidity and mortality worldwide. The aim of the present study was to determine the etiology of enteric viruses reported to the Catalan Microbiology Notification System (CMNS) (rotavirus and adenovirus 40-41) and to evaluate the importance of these viruses in enteric disease. Material and methods We retrospectively reviewed the notifications made from 1995-2002 by the laboratories of hospitals participating in the CMNS. Cases of acute gastroenteritis (AGE) were diagnosed by antigen detection in stools. The variables analyzed were: age, sex, month, and year of isolation. Statistical analysis was performed using the X2 test and Fisher's test and proportions were compared using the binomial test. The level of statistical significance was set at 0.05. Results Rotavirus was detected in 6708 cases of AGE (94.3%) and adenovirus 40-41 in 409 (5.7%). Viral AGE produced greater morbidity in children aged less than 2 years with 76% of cases caused by rotavi-rus and 68% of those caused by adenovirus 40-41 occurring in this age group. Rotavirus was isolated mainly in winter (68%), while adenovi-rus, although stable throughout the seasons, was more frequent in autumn (35.2%). Of cases of bacterial and viral enteritis detected by the CMNS, rotavirus was the second etiological agent and adenovirus the fourth in children aged less than 5 years after Campylobacter and Salmonella. Conclusions Of the various viruses that can cause AGE, some are highly frequent, such as calcivirus and astrovirus, which are not systematically detected and consequently the frequencies reported in the present study should be interpreted with caution. Nevertheless, rotavirus is the main cause of viral AGE, affecting small children, especially infants aged 1-11 months, with a clear predominance in winter. Adenovirus 40-41 mainly affects children aged less than 2 years and is more frequent in autumn. Vaccination against rotavirus should be a priority to reduce the high rates of morbidity and mortality caused by this disease.


title: Photons to illuminate the universe of sugar diversity through bioarrays

text: In this mini-review, we summarize the photochemical approaches for developing high-throughput carbohydrate microarray technologies. Newly established methods for photo-immobilizing unmodified monosaccharides, oligosaccharides and polysaccharides onto photoactive surfaces and coupling of photoactive carbohydrates onto polymer surfaces are reviewed.


title: Incidence, Risk Factors, and Outcomes of Idiopathic Pneumonia Syndrome after Allogeneic Hematopoietic Cell Transplantation

text: ABSTRACT Our current knowledge of idiopathic pneumonia syndrome (IPS) predates improved specificity in the diagnosis of IPS and advances in hematopoietic cell transplantation (HCT) and critical care practices. In this study, we describe and update the incidence, risk factors, and outcomes of IPS. We performed a retrospective cohort study of all adults who underwent allogeneic HCT at the Fred Hutchinson Cancer Research Center between 2006 and 2013 (n = 1829). IPS was defined using the National Heart, Lung, and Blood Institute consensus definition: multilobar airspace opacities on chest imaging, absence of lower respiratory tract infection, and hypoxemia. We described IPS incidence and mortality within 120 and 365 days after HCT. We examined conditioning intensity (nonmyeloablative versus myeloablative with high-dose total body irradiation [TBI] versus myeloablative with low-dose TBI) as an IPS risk factor in a time-to-event analysis using Cox models, controlled for age at transplant, HLA matching, stem cell source, and pretransplant Lung function Score (a combined measure of impairment in Forced Expiratory Volume in the first second (FEV1) and Diffusion capacity for carbon monoxide (DLCO)). Among 1829 HCT recipients, 67 fulfilled IPS criteria within 120 days (3.7%). Individuals who developed IPS were more likely to be black/non-Hispanic versus other racial groups and have severe pulmonary impairment but were otherwise similar to participants without IPS. In adjusted models, myeloablative conditioning with high-dose TBI was associated with increased risk of IPS (hazard ratio, 2.5; 95% confidence interval, 1.2 to 5.2). Thirty-one patients (46.3%) with IPS died within the first 120 days of HCT and 47 patients (70.1%) died within 365 days of HCT. In contrast, among the 1762 patients who did not acquire IPS in the first 120 days, 204 (11.6%) died within 120 days of HCT and 510 (29.9%) died within 365 days of HCT. Our findings suggest that although the incidence of IPS may be declining, it remains associated with post-transplant mortality. Future study should focus on early detection and identifying pathologic mediators of IPS to facilitate timely, targeted therapies for those most susceptible to lung injury post-HCT.


title: Endocrine changes in SARS-CoV-2 patients and lessons from SARS-CoV

text: Coronavirus infection outbreaks have occurred frequently in the last two decades and have led to significant mortality. Despite the focus on reducing mortality by preventing the spread of the virus, patients have died due to several other complications of the illness. The understanding of pathological mechanisms and their implications is continuously evolving. A number of symptoms occur in these patients due to the involvement of various endocrine glands. These clinical presentations went largely unnoticed during the first outbreak of severe acute respiratory syndrome (SARS) in 2002–2003. A few of these derangements continued during the convalescence phase and sometimes occurred after recovery. Similar pathological and biochemical changes are being reported with the novel coronavirus disease outbreak in 2020. In this review, we focus on these endocrine changes that have been reported in both SARS coronavirus and SARS coronavirus-2. As we battle the pandemic, it becomes imperative to address these underlying endocrine disturbances that are contributing towards or predicting mortality of these patients.


title: Understanding the Experience of an Extreme Event: A Personal Reflection

text: Mental models, which include our assumptions about how the world works, influence how we experience extreme events, such as the Australian bushfires. In turn, they can be altered by those experiences. Understanding (changes to) our mental models can help communities plan for, and recover from, extreme events.


title: Chapter 5 Acute Phase Proteins in Animals

text: Acute phase proteins (APP) were first identified in the early 1900s as early reactants to infectious disease. They are now understood to be an integral part of the acute phase response (APR) which is the cornerstone of innate immunity. APP have been shown to be valuable biomarkers as increases can occur with inflammation, infection, neoplasia, stress, and trauma. All animals—from fish to mammals—have demonstrable APP, but the type of major APP differs by species. While the primary application of these proteins in a clinical setting is prognostication, studies in animals have demonstrated relevance to diagnosis and detection and monitoring for subclinical disease. APP have been well documented in laboratory, companion, and large animals. With the advent of standardized and automated assays, these biomarkers are available for use in all fields of veterinary medicine as well as basic and clinical research.


title: Risk of severe COVID-19 in hypertensive patients treated with renin-angiotensin-aldosterone system inhibitors:

text: Abstract Introduction There is controversy concerning the use of angiotensin-converting enzyme inhibitors (ACEI) or angiotensin II type-I receptor blockers (ARB) for treating hypertensive patients with Covid-19. It has been hypothesized that these drugs might increase the risk of severe Covid-19, but some authors suggested that blocking the renin-angiotensin system might actually decrease this risk. Methods Retrospective cohort study of all the consecutive hypertensive patients with confirmed SARS-CoV-2 infection in a health area. The outcome variable was hospitalization because of severe Covid-19. Results 539 subjects were diagnosed of SARS-CoV-2 infection. Of these, 157 (29.1%) had hypertension and were included in the study. Sixty-nine cases (43.9%) were hospitalized because of severe Covid-19. In multivariable analysis older age, diabetes and hypertensive myocadiopathy were related to a higher risk of hospital admission. ARB treatment was associated with a significantly lower risk of hospitalization (HR: 0.29, 95% CI: 0.10 – 0.88). A similar albeit not significant trend was observed for ACEI. Conclusion ARB or ACEI treatment was not associated with a worse clinical outcome in consecutive hypertensive patients infected by SARS-CoV-2.


title: Thromboelastography Profiles of Critically Ill Patients With Coronavirus Disease 2019

text: OBJECTIVES: The rate of thromboembolic events among patients with coronavirus disease 2019 is high; however, there is no robust method to identify those at greatest risk. We reviewed thromboelastography studies in critically ill patients with coronavirus disease 2019 to characterize their coagulation states. DESIGN: Retrospective. SETTING: Tertiary ICU in New York City. PATIENTS: Sixty-four patients with coronavirus disease 2019 admitted to the ICU with thromboelastography performed. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Fifty percent of patients had a coagulation index in the hypercoagulable range (coagulation index > 3) (median 3.05). Reaction time and K values were below the lower limit of normal in 43.8% of the population consistent with a hypercoagulable profile. The median α angle and maximum amplitude (75.8° and 72.8 mm, respectively) were in the hypercoagulable range. The α angle was above reference range in 70.3% of patients indicative of rapid clot formation. Maximum amplitude, a factor of fibrinogen and platelet count and function, and a measure of clot strength was above reference range in 60.1% of patients. Thirty-one percent had thromboembolic events; thromboelastography parameters did not correlate with events in our cohort. Those with d-dimer values greater than 2,000 were more likely to have shorter reaction times compared with those with d-dimer levels less than or equal to 2,000 (4.8 vs 5.6 min; p = 0.001). CONCLUSIONS: A large proportion of critically ill patients with coronavirus disease 2019 have hypercoagulable thromboelastography profiles with additional derangements related to fibrinogen and platelet function. As the majority of patients have an elevated thromboelastography maximum amplitude, a follow-up study evaluating platelet aggregation would be instructive.


Modeling Marie Curie: How student nurses can contribute to evidence‐based practice during the COVID‐19 era


title: 2019 novel coronavirus disease (COVID-19) in Taiwan: Reports of two cases from Wuhan, China

text: Abstract We reported two cases with community-acquired pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) who returned from Wuhan, China in January, 2020. The reported cases highlight non-specific clinical presentations of 2019 novel coronavirus disease (COVID-19) as well as the importance of rapid laboratory-based diagnosis.


title: A novel model for protein sequence similarity analysis based on spectral radius

text: Abstract Advances in sequencing technologies led to rapid increase in the number and diversity of biological sequences, which facilitated development in the sequence research. In this paper, we present a new method for analyzing protein sequence similarity. We calculated the spectral radii of 20 amino acids (AAs) and put forward a novel 2-D graphical representation of protein sequences. To characterize protein sequences numerically, three groups of features were extracted and related to statistical, dynamics measurements and fluctuation complexity of the sequences. With the obtained feature vector, two models utilizing Gaussian Kernel similarity and Cosine similarity were built to measure the similarity between sequences. We applied our method to analyze the similarities/dissimilarities of four data sets. Both proposed models received consistent results with improvements when compared to that obtained by the ClustalW analysis. The novel approach we present in this study may therefore benefit protein research in medical and scientific fields.


title: Cyclophilin Inhibitors Restrict Middle East Respiratory Syndrome Coronavirus Via Interferon λ In Vitro And In Mice

text: RATIONALE: While severe coronavirus infections, including Middle East respiratory syndrome coronavirus (MERS-CoV) cause lung injury with high mortality rates, protective treatment strategies are not approved for clinical use. OBJECTIVES: We elucidated the molecular mechanisms by which the cyclophilin inhibitors Cyclosporin A (CsA) and Alisporivir (ALV) restrict MERS-CoV to validate their suitability as readily-available therapy in MERS-CoV infection. METHODS: Calu-3 cells and primary human alveolar epithelial cells (hAEC) were infected with MERS-CoV and treated with CsA or ALV or inhibitors targeting cyclophilin inhibitor-regulated molecules including Calcineurin, NFAT, or MAP kinases. Novel CsA-induced pathways were identified by RNA sequencing and manipulated by gene knockdown or neutralising antibodies. Viral replication was quantified by qRT-PCR and TCID(50). Data were validated in a murine MERS-CoV infection model. RESULTS: CsA and ALV both reduced MERS-CoV titers and viral RNA replication in Calu-3 and hAEC improving epithelial integrity. While neither Calcineurin nor NFAT inhibition reduced MERS-CoV propagation, blockade of c-Jun N-terminal kinase diminished infectious viral particle release but not RNA accumulation. Importantly, CsA induced interferon regulatory factor 1 (IRF1), a pronounced type-III-interferon (IFNλ) response and expression of antiviral genes. Down-regulation of IRF1 or IFNλ increased MERS-CoV propagation in presence of CsA. Importantly, oral application of CsA reduced MERS-CoV replication in vivo, correlating with elevated lung IFNλ levels and improved outcome. CONCLUSIONS: We provide evidence that cyclophilin inhibitors efficiently decrease MERS-CoV replication in vitro and in vivo via upregulation of inflammatory, antiviral cell responses, in particular IFNλ. CsA might therefore represent a promising candidate to treat MERS-CoV infection.


title: Chapter Eight Targeting the Channel Activity of Viroporins

text: Abstract Since the discovery that certain small viral membrane proteins, collectively termed as viroporins, can permeabilize host cellular membranes and also behave as ion channels, attempts have been made to link this feature to specific biological roles. In parallel, most viroporins identified so far are virulence factors, and interest has focused toward the discovery of channel inhibitors that would have a therapeutic effect, or be used as research tools to understand the biological roles of viroporin ion channel activity. However, this paradigm is being shifted by the difficulties inherent to small viral membrane proteins, and by the realization that protein–protein interactions and other diverse roles in the virus life cycle may represent an equal, if not, more important target. Therefore, although targeting the channel activity of viroporins can probably be therapeutically useful in some cases, the focus may shift to their other functions in following years. Small-molecule inhibitors have been mostly developed against the influenza A M2 (IAV M2 or AM2). This is not surprising since AM2 is the best characterized viroporin to date, with a well-established biological role in viral pathogenesis combined the most extensive structural investigations conducted, and has emerged as a validated drug target. For other viroporins, these studies are still mostly in their infancy, and together with those for AM2, are the subject of the present review.


title: Influenza and rhinovirus infections among health‐care workers

text: Background and objective: Health‐care workers (HCWs) are at higher risk of acquisition and transmission of respiratory virus infections. Nosocomial transmission of influenza has been documented but whether this is so for other respiratory viruses has not been assessed. Methods: Epidemiological, clinical and viral laboratory surveillance was carried out on HCWs presenting with acute respiratory infection in a university hospital. Results: Over a 2‐year period, 203 subjects were recruited: rhinovirus was the most frequently detected virus (37.7% in flu negative samples) and influenza A/B was positive in only 12.3% of subjects. Only 19.7% of HCWs were immunized against influenza. High detection of rhinovirus occurred even during the peak of the influenza season and half of the infected subjects reported an influenza‐like illness. Conclusion: Rhinovirus infection occurred frequently in this study population and probably contributes to influenza misdiagnosis. Educational interventions about different viruses causing respiratory symptoms and an increase in standards of infection control besides influenza immunization among HCWs is needed.


title: Functional analysis of the CC chemokine receptor 5 (CCR5) on virus-specific CD8+ T cells following coronavirus infection of the central nervous system

text: Abstract Intracranial infection of C57BL/6 mice with mouse hepatitis virus (MHV) results in an acute encephalomyelitis followed by a demyelinating disease similar in pathology to the human disease multiple sclerosis (MS). T cells participate in both defense and disease progression following MHV infection. Expression of chemokine receptors on activated T cells is important in allowing these cells to traffic into and accumulate within the central nervous system (CNS) of MHV-infected mice. The present study evaluated the contributions of CCR5 to the activation and trafficking of virus-specific CD8+ T cells into the MHV-infected CNS mice. Comparable numbers of virus-specific CD8+ T cells derived from immunized CCR5+/+ or CCR5−/− mice were present within the CNS of MHV-infected RAG1−/− mice following adoptive transfer, indicating that CCR5 is not required for trafficking of these cells into the CNS. RAG1−/− recipients of CCR5−/−-derived CD8+ T cells exhibited a modest, yet significant (P ≤ 0.05), reduction in viral burden within the brain which correlated with increased CTL activity and IFN-γ expression. Histological analysis of RAG1−/− recipients of either CCR5+/+or CCR5−/−-derived CD8+ T cells revealed only focal areas of demyelination with no significant differences in white matter destruction. These data indicate that CCR5 signaling on CD8+ T cells modulates antiviral activities but is not essential for entry into the CNS.


title: In Vivo Application of RNA Interference: From Functional Genomics to Therapeutics

text: Abstract RNAi has rapidly become a powerful tool for drug target discovery and validation in cell culture, and now has largely displaced efforts with antisense and ribozymes. Consequently, interest is rapidly growing for extension of its application to in vivo systems, such as animal disease models and human therapeutics. Studies on RNAi have resulted in two basic methods for its use for gene selective inhibition: 1) cytoplasmic delivery of short dsRNA oligonucleotides (siRNA), which mimics an active intermediate of an endogenous RNAi mechanism and 2) nuclear delivery of gene expression cassettes that express a short hairpin RNA (shRNA), which mimics the micro interfering RNA (miRNA) active intermediate of a different endogenous RNAi mechanism. Non‐viral gene delivery systems are a diverse collection of technologies that are applicable to both of these forms of RNAi. Importantly, unlike antisense and ribozyme systems, a remarkable trait of siRNA is a lack of dependence on chemical modifications blocking enzymatic degradation, although chemical protection methods developed for the earlier systems are being incorporated into siRNA and are generally compatible with non‐viral delivery systems. The use of siRNA is emerging more rapidly than for shRNA, in part due to the increased effort required to construct shRNA expression systems before selection of active sequences and verification of biological activity are obtained. In contrast, screens of many siRNA sequences can be accomplished rapidly using synthetic oligos. It is not surprising that the use of siRNA in vivo is also emerging first. Initial in vivo studies have been reported for both viral and non‐viral delivery but viral delivery is limited to shRNA. This review describes the emerging in vivo application of non‐viral delivery systems for RNAi for functional genomics, which will provide a foundation for further development of RNAi therapeutics. Of interest is the rapid adaptation of ligand‐targeted plasmid‐based nanoparticles for RNAi agents. These systems are growing in capabilities and beginning to pose a serious rival to viral vector based gene delivery. The activity of siRNA in the cytoplasm may lower the hurdle and thereby accelerate the successful development of therapeutics based on targeted non‐viral delivery systems.


title: The Application of the Haddon Matrix to Public Health Readiness and Response Planning

text: State and local health departments continue to face unprecedented challenges in preparing for, recognizing, and responding to threats to the public’s health. The attacks of 11 September 2001 and the ensuing anthrax mailings of 2001 highlighted the public health readiness and response hurdles posed by intentionally caused injury and illness. At the same time, recent natural disasters have highlighted the need for comparable public health readiness and response capabilities. Public health readiness and response activities can be conceptualized similarly for intentional attacks, natural disasters, and human-caused accidents. Consistent with this view, the federal government has adopted the all-hazards response model as its fundamental paradigm. Adoption of this paradigm provides powerful improvements in efficiency and efficacy, because it reduces the need to create a complex family of situation-specific preparedness and response activities. However, in practice, public health preparedness requires additional models and tools to provide a framework to better understand and prioritize emergency readiness and response needs, as well as to facilitate solutions; this is particularly true at the local health department level. Here, we propose to extend the use of the Haddon matrix—a conceptual model used for more than two decades in injury prevention and response strategies—for this purpose.


title: Synthesis and evaluation of pyrazolone compounds as SARS-coronavirus 3C-like protease inhibitors

text: Abstract A series of pyrazolone compounds as possible SARS-CoV 3CL protease inhibitors were designed, synthesized, and evaluated by in vitro protease assay using fluorogenic substrate peptide in which several showed potent inhibition against the 3CL protease. Interestingly, one of the inhibitors was also active against 3C protease from coxsackievirus B3. These inhibitors could be potentially developed into anti-coronaviral and anti-picornaviral agents.


title: Seizures associated with coronavirus infections

text: Neurotropic and neuroinvasive capabilities of coronaviruses have been described in humans. Neurological problems found in patients with coronavirus infection include: febrile seizures, convulsions, loss of consciousness, encephalomyelitis, and encephalitis. Coronavirus disease (COVID-19) is caused by SARS-COV2. In severe cases, patients may develop severe pneumonia, acute respiratory distress syndrome, and acute cardiac injury. While seizures and status epilepticus have not been widely reported in the past five months since the onset of COVID-19 pandemic, patients with COVID-19 may have hypoxia, multiorgan failure, and severe metabolic and electrolyte disarrangements; hence, it is plausible to expect clinical or subclinical acute symptomatic seizures to happen in these patients. One should be prepared to treat seizures appropriately, if they happen in a patient who is already in a critical medical condition and suffers from organ failure.


Circulation of influenza A viruses in dromedary camels in Saudi Arabia


title: Inhibition of replication and transcription activator and latency-associated nuclear antigen of Kaposi's sarcoma-associated herpesvirus by morpholino oligomers

text: Abstract Kaposi's sarcoma-associated herpesvirus (KSHV) is associated with Kaposi's sarcoma and primary effusion lymphoma (PEL). The KSHV replication and transcription activator (RTA) and latency-associated nuclear antigen (LANA) play key roles in activating KSHV lytic replication and maintaining KSHV latency, respectively. Phosphorodiamidate morpholino oligomers (PMO) are similar to short single-stranded DNA oligomers, but possess a modified backbone that confers highly specific binding and resistance to nucleases. In this study, RTA and LANA mRNA in PEL cells were targeted by antisense peptide-conjugated PMO (P-PMO) in an effort to suppress KSHV replication. Highly efficient P-PMO uptake by PEL cells was observed. Treatment of PEL cells with a RTA P-PMO (RP1) reduced RTA expression in a dose-dependent and sequence-specific manner, and also caused a significant decrease in several KSHV early and late gene products, including vIL-6, vIRF-1, and ORF-K8.1A. KSHV viral DNA levels were reduced both in cells and culture supernatants of RP1 P-PMO-treated cells, indicating that KSHV lytic replication was suppressed. Treatment of BCBL-1 cells with P-PMO against LANA resulted in a reduction of LANA expression. Cell viability assays detected no cytotoxicity from P-PMO alone, within the concentration range used for the experiments in this study. These results suggest that RP1 P-PMO can specifically block KSHV replication, and further study is warranted.


title: Protein Sequence Comparison and DNA-binding Protein Identification with Generalized PseAAC and Graphical Representation

text: AIM AND OBJECTIVE: The rapid increase in the amount of protein sequence data available leads to an urgent need for novel computational algorithms to analyze and compare these sequences. This study is undertaken to develop an efficient computational approach for timely encoding protein sequences and extracting the hidden information. METHODS: Based on two physicochemical properties of amino acids, a protein primary sequence was converted into a three-letter sequence, and then a graph without loops and multiple edges and its geometric line adjacency matrix were obtained. A generalized PseAAC (pseudo amino acid composition) model was thus constructed to characterize a protein sequence numerically. RESULTS: By using the proposed mathematical descriptor of a protein sequence, similarity comparisons among β-globin proteins of 17 species and 72 spike proteins of coronaviruses were made, respectively. The resulting clusters agreed well with the established taxonomic groups. In addition, a generalized PseAAC based SVM (support vector machine) model was developed to identify DNA-binding proteins. Experiment results showed that our method performed better than DNAbinder, DNA-Prot, iDNA-Prot and enDNA-Prot by 3.29-10.44% in terms of ACC, 0.056-0.206 in terms of MCC, and 1.45-15.76% in terms of F1M. When the benchmark dataset was expanded with negative samples, the presented approach outperformed the four previous methods with improvement in the range of 2.49-19.12% in terms of ACC, 0.05-0.32 in terms of MCC, and 3.82-33.85% in terms of F1M. CONCLUSION: These results suggested that the generalized PseAAC model was very efficient for comparison and analysis of protein sequences, and very competitive in identifying DNA-binding proteins.


title: Can commonly-used fan-driven air cleaning technologies improve indoor air quality? A literature review

text: Abstract Air cleaning techniques have been applied worldwide with the goal of improving indoor air quality. The effectiveness of applying these techniques varies widely, and pollutant removal efficiency is usually determined in controlled laboratory environments which may not be realized in practice. Some air cleaners are largely ineffective, and some produce harmful by-products. To summarize what is known regarding the effectiveness of fan-driven air cleaning technologies, a state-of-the-art review of the scientific literature was undertaken by a multidisciplinary panel of experts from Europe, North America, and Asia with expertise in air cleaning, aerosol science, medicine, chemistry and ventilation. The effects on health were not examined. Over 26,000 articles were identified in major literature databases; 400 were selected as being relevant based on their titles and abstracts by the first two authors, who further reduced the number of articles to 160 based on the full texts. These articles were reviewed by the panel using predefined inclusion criteria during their first meeting. Additions were also made by the panel. Of these, 133 articles were finally selected for detailed review. Each article was assessed independently by two members of the panel and then judged by the entire panel during a consensus meeting. During this process 59 articles were deemed conclusive and their results were used for final reporting at their second meeting. The conclusions are that: (1) None of the reviewed technologies was able to effectively remove all indoor pollutants and many were found to generate undesirable by-products during operation. (2) Particle filtration and sorption of gaseous pollutants were among the most effective air cleaning technologies, but there is insufficient information regarding long-term performance and proper maintenance. (3) The existing data make it difficult to extract information such as Clean Air Delivery Rate (CADR), which represents a common benchmark for comparing the performance of different air cleaning technologies. (4) To compare and select suitable indoor air cleaning devices, a labeling system accounting for characteristics such as CADR, energy consumption, volume, harmful by-products, and life span is necessary. For that purpose, a standard test room and condition should be built and studied. (5) Although there is evidence that some air cleaning technologies improve indoor air quality, further research is needed before any of them can be confidently recommended for use in indoor environments.


title: Distribution of bat-borne viruses and environment patterns

text: Abstract Environmental modifications are leading to biodiversity changes, loss and habitat disturbance. This in turn increases contacts between wildlife and hence the risk of transmission and emergence of zoonotic diseases. We analyzed the environment and land use using remote spatial data around the sampling locations of bats positive for coronavirus (21 sites) and astrovirus (11 sites) collected in 43 sites. A clear association between viruses and hosts was observed. Viruses associated to synanthropic bat genera, such as Myotis or Scotophilus were associated to highly transformed habitats with human presence while viruses associated to fruit bat genera were correlated with natural environments with dense forest, grassland areas and regions of high elevation. In particular, group C betacoronavirus were associated with mosaic habitats found in anthropized environments.


title: Composition and Three-Dimensional Architecture of the Dengue Virus Replication and Assembly Sites

text: Positive-strand RNA viruses are known to rearrange cellular membranes to facilitate viral genome replication. The biogenesis and three-dimensional organization of these membranes and the link between replication and virus assembly sites is not fully clear. Using electron microscopy, we find Dengue virus (DENV)-induced vesicles, convoluted membranes, and virus particles to be endoplasmic reticulum (ER)-derived, and we detect double-stranded RNA, a presumed marker of RNA replication, inside virus-induced vesicles. Electron tomography (ET) shows DENV-induced membrane structures to be part of one ER-derived network. Furthermore, ET reveals vesicle pores that could enable release of newly synthesized viral RNA and reveals budding of DENV particles on ER membranes directly apposed to vesicle pores. Thus, DENV modifies ER membrane structure to promote replication and efficient encapsidation of the genome into progeny virus. This architecture of DENV replication and assembly sites could explain the coordination of distinct steps of the flavivirus replication cycle.


title: Polyprotein Processing as a Determinant for In Vitro Activity of Semliki Forest Virus Replicase

text: Semliki Forest virus (SFV) is an arthropod-borne alphavirus that induces membrane invaginations (spherules) in host cells. These harbor the viral replication complexes (RC) that synthesize viral RNA. Alphaviruses have four replicase or nonstructural proteins (nsPs), nsP1–4, expressed as polyprotein P1234. An early RC, which synthesizes minus-strand RNA, is formed by the polyprotein P123 and the polymerase nsP4. Further proteolytic cleavage results in a late RC consisting of nsP1–4 and synthesizing plus strands. Here, we show that only the late RCs are highly active in RNA synthesis in vitro. Furthermore, we demonstrate that active RCs can be isolated from both virus-infected cells and cells transfected with the wild-type replicase in combination with a plasmid expressing a template RNA. When an uncleavable polyprotein P123 and polymerase nsP4 were expressed together with a template, high levels of minus-strand RNA were produced in cells, but RCs isolated from these cells were hardly active in vitro. Furthermore, we observed that the uncleavable polyprotein P123 and polymerase nsP4, which have previously been shown to form spherules even in the absence of the template, did not replicate an exogenous template. Consequently, we hypothesize that the replicase proteins were sequestered in spherules and were no longer able to recruit a template.