title: In vivo functional characterization of the SARS-Coronavirus 3a protein in Drosophila

text: Abstract The Severe Acute Respiratory Syndrome-Coronavirus (SARS-CoV) 3a locus encodes a 274 a.a. novel protein, and its expression has been confirmed in SARS patients. To study functional roles of 3a, we established a transgenic fly model for the SARS-CoV 3a gene. Misexpression of 3a in Drosophila caused a dominant rough eye phenotype. Using a specific monoclonal antibody, we demonstrated that the 3a protein displayed a punctate cytoplasmic localization in Drosophila as in SARS-CoV-infected cells. We provide genetic evidence to support that 3a is functionally related to clathrin-mediated endocytosis. We further found that 3a misexpression induces apoptosis, which could be modulated by cellular cytochrome c levels and caspase activity. From a forward genetic screen, 78 dominant 3a modifying loci were recovered and the identity of these modifiers revealed that the severity of the 3a-induced rough eye phenotype depends on multiple cellular processes including gene transcriptional regulation.


title: Chapter Four Arthropod-Borne Flaviviruses and RNA Interference Seeking New Approaches for Antiviral Therapy

text: Abstract Flaviviruses are the most prevalent arthropod-borne viruses worldwide, and nearly half of the 70 Flavivirus members identified are human pathogens. Despite the huge clinical impact of flaviviruses, there is no specific human antiviral therapy available to treat infection with any of the flaviviruses. Therefore, there is a continued search for novel therapies, and this review describes the current knowledge on the usage of RNA interference (RNAi) in combating flavivirus infections. RNAi is a process of sequence-specific gene silencing triggered by double-stranded RNA. Antiviral RNAi strategies against arthropod-borne flaviviruses have been reported and although several hurdles must be overcome to employ this technology in clinical applications, they potentially represent a new therapeutic tool.


title: Discovery of Small-Molecule HIV-1 Fusion and Integrase Inhibitors Oleuropein and Hydroxytyrosol: I. Fusion Inhibition

text: We have identified oleuropein (Ole) and hydroxytyrosol (HT) as a unique class of HIV-1 inhibitors from olive leaf extracts effective against viral fusion and integration. We used molecular docking simulation to study the interactions of Ole and HT with viral targets. We find that Ole and HT bind to the conserved hydrophobic pocket on the surface of the HIV-gp41 fusion domain by hydrogen bonds with Q577 and hydrophobic interactions with I573, G572, and L568 on the gp41 N-terminal heptad repeat peptide N36, interfering with formation of the gp41 fusion-active core. To test and confirm modeling predications, we examined the effect of Ole and HT on HIV-1 fusion complex formation using native polyacrylamide gel electrophoresis and circular dichroism spectroscopy. Ole and HT exhibit dose dependent inhibition on HIV-1 fusion core formation with EC(50)s of 66–58 nM, with no detectable toxicity. Our findings on effects on HIV-1 integrase are reported separately.


Influenza A and B Viruses but Not MERS-CoV in Hajj Pilgrims, Austria, 2014


title: Current Precautions and Future Directions in Lung Transplantation during the COVID‐19 Pandemic

text: The unprecedented public health emergency caused by the acute viral respiratory coronavirus disease (COVID‐19) has drastically changed current practices in solid organ transplantation, markedly so for transplantation of the lungs, the major target of the virus. Although national and state authorities do not recommend postponing transplant procedures, most specialists are reluctant to proceed due to substantial uncertainty and increased risks in the midst of the pandemic. There is an urgent need for evidence‐based directions to move forward. Here, we offer our insights as specialists at a high‐volume center located in a geographical area with high infection rates.


title: Zinc Iodide in combination with Dimethyl Sulfoxide for treatment of SARS-CoV-2 and other viral infections

text: Zinc Iodide and Dimethyl Sulfoxide composition is proposed as therapeutic agents to treat and prevent chronic and acute viral infections including SARS-CoV-2 infected patients. The therapeutic combinations have a wide range of virucidal effect on DNA and RNA containing viruses. The combination also exhibits anti-inflammatory, immunomodulating, antifibrotic, antibacterial, antifungal and antioxidative effects. Given the fact that Zinc Iodide has been used as an oral antiseptic agent and DMSO have already proven as a safe pharmaceutical solvent and therapeutic agent, we hypothesize that the combination of these two agents can be applied as an effective, safe and inexpensive treatment for SARS-CoV-2 and other viral infection. The therapeutic compound can be applied as both etiological and pathogenesis therapy and used as an effective and safe antiseptic (disinfectant) for human and animals as well.


Living in pandemic times


title: Metagenomic analysis of viral genetic diversity in respiratory samples from children with severe acute respiratory infection in China

text: Abstract Severe acute respiratory infection (SARI) in children is thought to be mainly caused by infection with various viruses, some of which have been well characterized; however, analyses of respiratory tract viromes among children with SARI versus those without are limited. In this study, nasopharyngeal swabs from children with and without SARI (135 versus 15) were collected in China between 2008 and 2010 and subjected to multiplex metagenomic analyses using a next-generation sequencing platform. The results show that members of the Paramyxoviridae, Coronaviridae, Parvoviridae, Orthomyxoviridae, Picornaviridae, Anelloviridae and Adenoviridae families represented the most abundant species identified (>50% genome coverage) in the respiratory tracts of children with SARI. The viral population found in the respiratory tracts of children without SARI was less diverse and mainly dominated by the Anelloviridae family with only a small proportion of common epidemic respiratory viruses. Several almost complete viral genomes were assembled, and the genetic diversity was determined among several samples based on next-generation sequencing. This research provides comprehensive mapping of the viromes of children with SARI and indicates high heterogeneity of known viruses present in the childhood respiratory tract, which may benefit the detection and prevention of respiratory disease.


title: Digitally aided telemedicine during the SARS‐CoV‐2 pandemic to screen oral medicine emergencies

text: A new human coronavirus, known as severe acute respiratory coronavirus 2 syndrome (SARS‐CoV‐2), emerged at the end of 2019 in Wuhan, China, and we are now facing a pandemic (WHO 2020). The virus has been found in saliva (Meng et al 2020, Sabino et al 2020). In vitro experimental findings suggest that SARS‐CoV‐2 remained viable in aerosols for 3 hours (van Doremalen et al 2020).


title: “Monoclonal-Type” Plastic Antibodies for COVID-19 Treatment: What Is the Idea?

text: In late December 2019, an outbreak due to a novel coronavirus, initially called 2019-nCoV, was reported in Wuhan, China [...].


title: Immune responses in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV)

text: Abstract In three successive experiments, the immune functions of pigs persistently infected with the porcine reproductive and respiratory syndrome virus (PRRSV) have been evaluated. Non-specific immune responses were analyzed over a period of 12 weeks post-infection (PI). In addition, the capacity of PRRSV-infected pigs to develop an efficient immune response against pseudorabies virus (PRV) glycoproteins and to resist to a subsequent virulent challenge was investigated. Our results demonstrate that PRRSV produced minor effects on the immune system of pigs. The skin delayed type hypersensitivity (DTH) in response to phytohemagglutinine injection was slightly diminished one week after challenge, but was restored thereafter. However, three weeks after the infection, the total white blood cell count, and the number of CD2+, CD8+ and IgM+ cells were enhanced. The increase in numbers of CD8+ cells persisted for three consecutive weeks. Serum immunoglobulins in infected pigs also increased by week 3 PI and up to 8 weeks PI. These results show that PRRSV may have stimulating effects on the pig immune system during the phase of long-lasting infection. After immunization with PRV glycoproteins, the production of anti-PRV antibodies and skin DTH response against PRV glycoproteins were not affected. On the contrary, following a virulent PRV challenge, PRRSV-infected pigs developed a better secondary antibody response and their resistance to the infection was as effective as in control pigs. Taken together, our data do not support a systemic immunosuppressive effect of PRRSV, during the persistent phase of infection. Other mechanisms may therefore apply to explain the emergence of secondary infections in endemically infected herds.


title: Analyzing the impact of the coronavirus crisis on business models

text: Abstract In the light of the current coronavirus crisis, business-to-business firms face a variety of challenges in a complex and fast-changing environment. In order to provide structured analysis and to guide strategic decision-making, we present a novel, five-step approach for analyzing the impact of a crisis on a firm's business model. We applied the approach with eight business-to-business firms and find support for its usefulness. The evidence suggests very different impacts of the coronavirus crisis on business-to-business firms, and that understanding these differences is important for strategizing during the crisis but also to navigating successfully into the future. We also describe six different types of crisis impacts on business models. We conclude by developing managerial implications and questions for future research.


title: Baculovirus Surface Display of Zika Virus Envelope Protein Protects against Virus Challenge in Mouse Model

text: Zika virus (ZIKV) is emerging as a significant pathogen worldwide and may cause severe neurological disorders such as fetal microcephaly and Guillain-Barre syndrome. No drug or listed vaccines are currently available for preventing ZIKV infection. As a major target of neutralizing, ZIKV envelop (E) protein usually used for vaccine development. Nevertheless, the immunogenicity of ZIKV envelop (E) protein expressed by baculovirus display system has never been assessed. In this study, we reported a new strategy for surface display of ZIKV E protein by a recombinant baculovirus vector derived from Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) and assessed its immunogenicity in mice. We produced recombinant fusion ZIKV E protein linked with signal peptide (SP) and transmembrane domain (TM) of AcMNPV GP64. The results showed that the recombinant protein was easy to produce by baculovirus display system. BALB/c mice immunized with this recombinant E protein developed ZIKV specific serum antibodies. The anti-E protein sera from the mice were able to effectively neutralize ZIKV in vitro. More importantly, AG6 (IFN-α/β and IFN-γ receptor deficient) mice immunized with recombinant E protein were protected against lethal ZIKV challenge. Together, these findings demonstrated that the recombinant E protein displayed by baculovirus can be conveniently prepared and displayed good immunogenicity in immunized mice. It is a promising practical approach for prompting the development of vaccine and related immunology research. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12250-020-00238-x) contains supplementary material, which is available to authorized users.


title: Lactic acid bacteria: reviewing the potential of a promising delivery live vector for biomedical purposes

text: Lactic acid bacteria (LAB) have a long history of safe exploitation by humans, being used for centuries in food production and preservation and as probiotic agents to promote human health. Interestingly, some species of these Gram-positive bacteria, which are generally recognized as safe organisms by the US Food and Drug Administration (FDA), are able to survive through the gastrointestinal tract (GIT), being capable to reach and colonize the intestine, where they play an important role. Besides, during the last decades, an important effort has been done for the development of tools to use LAB as microbial cell factories for the production of proteins of interest. Given the need to develop effective strategies for the delivery of prophylactic and therapeutic molecules, LAB have appeared as an appealing option for the oral, intranasal and vaginal delivery of such molecules. So far, these genetically modified organisms have been successfully used as vehicles for delivering functional proteins to mucosal tissues in the treatment of many different pathologies including GIT related pathologies, diabetes, cancer and viral infections, among others. Interestingly, the administration of such microorganisms would suppose a significant decrease in the production cost of the treatments agents since being live organisms, such vectors would be able to autonomously amplify and produce and deliver the protein of interest. In this context, this review aims to provide an overview of the use of LAB engineered as a promising alternative as well as a safety delivery platform of recombinant proteins for the treatment of a wide range of diseases.


title: Self-Assessment Questionnaire for Efficient and Safe Evaluation of Patients with Mild COVID-19

text: As the outbreak of coronavirus disease 2019 continues and the number of confirmed cases requiring isolation increases, there is a need for a safe and efficient system to assess patients' condition. We developed and evaluated a self-assessment questionnaire consisting of 23 symptoms with linear-scale scores from 0 to 10. Patients were asked to indicate their worst score for each symptom daily, and medical personnel assessed clinical improvement or deterioration based on the changes in scores. Focused communication on severity of specific symptoms was the primary advantage for the clinicians, and a thorough check for their symptoms was helpful for patients.


title: NetNCSP: Nonoverlapping closed sequential pattern mining

text: Abstract Sequential pattern mining (SPM) has been applied in many fields. However, traditional SPM neglects the pattern repetition in sequence. To solve this problem, gap constraint SPM was proposed and can avoid finding too many useless patterns. Nonoverlapping SPM, as a branch of gap constraint SPM, means that any two occurrences cannot use the same sequence letter in the same position as the occurrences. Nonoverlapping SPM can make a balance between efficiency and completeness. The frequent patterns discovered by existing methods normally contain redundant patterns. To reduce redundant patterns and improve the mining performance, this paper adopts the closed pattern mining strategy and proposes a complete algorithm, named Nettree for Nonoverlapping Closed Sequential Pattern (NetNCSP) based on the Nettree structure. NetNCSP is equipped with two key steps, support calculation and closeness determination. A backtracking strategy is employed to calculate the nonoverlapping support of a pattern on the corresponding Nettree, which reduces the time complexity. This paper also proposes three kinds of pruning strategies, inheriting, predicting, and determining. These pruning strategies are able to find the redundant patterns effectively since the strategies can predict the frequency and closeness of the patterns before the generation of the candidate patterns. Experimental results show that NetNCSP is not only more efficient but can also discover more closed patterns with good compressibility. Furtherly, in biological experiments NetNCSP mines the closed patterns in SARS-CoV-2 and SARS viruses. The results show that the two viruses are of similar pattern composition with different combinations.


Binding stretches of myelin basic protein define encephalitogenic T cell epitopes for RT1.B and RT1.D alleles of four different rat haplotypes


title: Low dose lung radiotherapy for COVID-19 pneumonia. The rationale for a cost-effective anti-inflammatory treatment

text: The COVID-19 pandemia is affecting people worldwide. Most of the patients suffered of a respiratory disease that will progress to an acute respiratory distress syndrome (ARDS). SARS-CoV-2 pneumonia severely ill patients, develop a systemic inflammatory response with a Cytokine Release Syndrome (CRS), that is characterized by a sudden increase in several pro-inflammatory cytokines, mainly IL-1, IL-6 and TNF-alfa by activated macrophages (M1 phenotype). Blocking IL-6 with tocilizumab and using respirator equipment seems to be a very important issue in this (SARS-CoV-2) pneumonia, but not all patients are referred to such treatments. Low dose radiotherapy (0,5 Gy), is an evidence-based anti-inflammatory treatment, that could modify the immune landscape in the lung affected of SARS-CoV-2 pneumonia, through macrophages polarization to alternatively activated Macrophages (M2 phenotype). Radiation-induced cancer risk could be assumed due to the very low dose used, the advanced age of the patients and the life-threatening condition of SARS-Cov2 pneumonia. LDRT is a cost-effective non-toxic treatment already available in most general hospitals. This fact allows that it would be used for the large number of patients that will suffer this disease, and that would not receive specific anti-IL-6 treatments in ICUs in low and middle income countries.


title: Acute myocarditis associated with novel Middle East respiratory syndrome coronavirus

text: The novel Middle East respiratory syndrome coronavirus (MERS-CoV) has been identified as a cause of pneumonia; however, it has not been reported as a cause of acute myocarditis. A 60-year-old man presented with pneumonia and congestive heart failure. On the first day of admission, he was found to have an elevated troponin-I level and severe global left ventricular systolic dysfunction on echocardiography. The serum creatinine level was found mildly elevated. Chest radiography revealed in the lower lung fields accentuated bronchovascular lung markings and multiple small patchy opacities. Laboratory tests were negative for viruses known to cause myocarditis. Sputum sample was positive for MERS-CoV. Cardiovascular magnetic resonance revealed evidence of acute myocarditis. The patient had all criteria specified by the International Consensus Group on CMR in Myocarditis that make a clinical suspicion for acute myocarditis. This was the first case that demonstrated that MERS-CoV may cause acute myocarditis and acute-onset heart failure.


title: Infectious Disease Surveillance

text: World Health Organization retains copyright in the manuscript and provides Elsevier the permission to publish the manuscript as a chapter in this book.


title: Advances in Real‐Time PCR: Application to Clinical Laboratory Diagnostics

text: The polymerase chain reaction (PCR) has become one of the most important tools in molecular diagnostics, providing exquisite sensitivity and specificity for detection of nucleic acid targets. Real-time monitoring of PCR has simplified and accelerated PCR laboratory procedures and has increased information obtained from specimens including routine quantification and differentiation of amplification products. Clinical diagnostic applications and uses of real-time PCR are growing exponentially, real-time PCR is rapidly replacing traditional PCR, and new diagnostic uses likely will emerge. This review analyzes the scope of present and potential future clinical diagnostic applications of this powerful technique. Critical discussions focus on basic concepts, variations, data analysis, instrument platforms, signal detection formats, sample collection, assay design, and execution of real-time PCR.


title: Emotional distress in young adults during the COVID-19 pandemic: evidence of risk and resilience from a longitudinal cohort study

text: BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic and associated lockdown could be considered a ‘perfect storm’ for increases in emotional distress. Such increases can only be identified by studies that use data collected before and during the pandemic. Longitudinal data are also needed to examine (1) the roles of previous distress and stressors in emotional distress during the pandemic and (2) how COVID-19-related stressors and coping strategies are associated with emotional distress when pre-pandemic distress is accounted for. METHODS: Data came from a cohort study (N = 768). Emotional distress (perceived stress, internalizing symptoms, and anger), COVID-19-related stressors, and coping strategies were measured during the pandemic/lockdown when participants were aged 22. Previous distress and stressors were measured before COVID-19 (at age 20). RESULTS: On average, participants showed increased levels of perceived stress and anger (but not internalizing symptoms) during the pandemic compared to before. Pre-COVID-19 emotional distress was the strongest predictor of during-pandemic emotional distress, followed by during-pandemic economic and psychosocial stressors (e.g. lifestyle and economic disruptions) and hopelessness, and pre-pandemic social stressors (e.g. bullying victimization and stressful life events). Most health risks to self or loved ones due to COVID-19 were not uniquely associated with emotional distress in final models. Coping strategies associated with reduced distress included keeping a daily routine, physical activity, and positive reappraisal/reframing. CONCLUSIONS: In our community sample, pre-pandemic distress, secondary consequences of the pandemic (e.g. lifestyle and economic disruptions), and pre-pandemic social stressors were more consistently associated with young adults' emotional distress than COVID-19-related health risk exposures.


title: Advanced removal of C. famata in bioaerosols by simultaneous adsorption and photocatalytic oxidation of Cu-doped TiO2/PU under visible irradiation

text: Abstract Polyurethane (PU), a honeycomb structure material, was used as a substrate onto which TiO2 and Cu–TiO2 were deposited in order to integrate the adsorption property to the photocatalysts. TiO2 deposited on PU (TiO2/PU) and Cu-doped TiO2 deposited on PU (Cu–TiO2/PU) were synthesized and applied to the removal of Candida famata (C. famata), a frequently encountered airborne yeast. The removal capacities of C. famata by PU, TiO2/PU and Cu–TiO2/PU were 1.5×105, 3.2×105 and 6.9×105 (CFU/cm3) under dark condition and 1.5×105, 3.3×105 and 1.8×106 (CFU/cm3) under visible light irradiation, respectively. PU and TiO2/PU seemed to exhibit only an adsorption ability for removing C. famata in aerosols under both dark and visible light. The C. famata removal capacity of Cu–TiO2/PU under visible light was increased 2.6-fold compared to that under dark condition. This significant increase was attributed to the Cu dopant, which enhanced the electron–hole separation efficiency and capacity of TiO2, resulting in the high photocatalytic activity of Cu–TiO2/PU under visible light.


title: Nontraumatic Osteonecrosis of the Distal Tibia: A Case Presentation and Review of the Literature

text: Abstract Osteonecrosis, although commonly occurring in the hip, can also affect the leg and foot. In the foot, it most commonly occurs in the talus. The incidence of osteonecrosis occurring in the tibia is relatively rare. We report a case of a woman who presented to our clinic with ankle pain that was idiopathic in nature. Subsequent magnetic resonance imaging showed findings consistent with osteonecrosis of the bilateral distal tibias and several other lesions located in the shoulder, hip, and calcaneus. The present report also serves as a review of both etiology and treatment of osteonecrosis as it relates to the lower extremity.


title: Acute Necrotizing Pancreatitis Associated with Mycoplasma pneumoniae Infection in a Child

text: Mycoplasma pneumoniae is responsible for approximately 20% to 30% of community-acquired pneumonia, and is well known for its diverse extrapulmonary manifestations. However, acute necrotizing pancreatits is an extremely rare extrapulmonary manifestation of M. pneumoniae infection. A 6-year-old girl was admitted due to abdominal pain, vomiting, fever, and confused mentality. Acute necrotizing pancreatitis was diagnosed according to symptoms, laboratory test results, and abdominal computed tomography scans. M. pneumoniae infection was diagnosed by a 4-fold increase in antibodies to M. pneumoniae between acute and convalescent sera by particle agglutination antibody assay. No other etiologic factors or pathogens were detected. Despite the occurrence of a large infected pseudocyst during the course, the patient was able to discharge without morbidity by early aggressive supportive care. This is the first case in Korea of a child with acute necrotizing pancreatitis associated with M. pneumoniae infection.


title: Gonococci cause immunosuppression by engaging a coinhibitory receptor on T lymphocytes

text: Gonococci that bind the coinhibitory receptor CEACAM1 appear to down-regulate the activation and proliferation of CD4(+) T cells. Such infection-induced immunosuppression helps explain why there is little specific immune response associated with gonococcal disease.


title: Human choline dehydrogenase: Medical promises and biochemical challenges

text: Abstract Human choline dehydrogenase (CHD) is located in the inner membrane of mitochondria primarily in liver and kidney and catalyzes the oxidation of choline to glycine betaine. Its physiological role is to regulate the concentrations of choline and glycine betaine in the blood and cells. Choline is important for regulation of gene expression, the biosynthesis of lipoproteins and membrane phospholipids and for the biosynthesis of the neurotransmitter acetylcholine; glycine betaine plays important roles as a primary intracellular osmoprotectant and as methyl donor for the biosynthesis of methionine from homocysteine, a required step for the synthesis of the ubiquitous methyl donor S-adenosyl methionine. Recently, CHD has generated considerable medical attention due to its association with various human pathologies, including male infertility, homocysteinuria, breast cancer and metabolic syndrome. Despite the renewed interest, the biochemical characterization of the enzyme has lagged behind due to difficulties in the obtainment of purified, active and stable enzyme. This review article summarizes the medical relevance and the physiological roles of human CHD, highlights the biochemical knowledge on the enzyme, and provides an analysis based on the comparison of the protein sequence with that of bacterial choline oxidase, for which structural and biochemical information is available.


title: Th2 predominance and CD8+ memory T cell depletion in patients with severe acute respiratory syndrome

text: Abstract The immune spectrum of severe acute respiratory syndrome (SARS) is poorly understood. To define the dynamics of the immune spectrum in SARS, serum levels of cytokines, chemokines, immunoglobulins, complement and specific antibodies against SARS-associated coronavirus (SARS-CoV) were assayed by enzyme-linked immunosorbent assay (ELISA), and phenotypes of peripheral lymphocytes were analyzed by flow cytometry in 95 SARS-infected patients. Results showed that interleukin (IL)-10 and transforming growth factor β (TGF-β) were continuously up-regulated during the entirety of SARS. Regulated on activation normally T cell-expressed and secreted (RANTES) levels were decreased, while monocyte chemoattractant protein-1 (MCP-1) was elevated in acute patients. Immunoglobulins and complement were elevated during the first month of SARS. Both serum-positive rates and titers of specific IgM and IgG antibodies responding to SARS-CoV peaked at days 41–60 from the onset of SARS. CD4+ and CD8+ T lymphocytes decreased significantly in acute-phase. CD3+CD8+CD45RO+ T lymphocytes were decreased by 36.78% in the convalescent patients. Conclusion: SARS-CoV seemed to elicit effective humoral immunity but inhibited cellular immunity, especially CD8+ memory T lymphocytes over time. Prolonged overproduction of IL-10 and TGF-β may play an important role in the disease.


title: Experience of using convalescent plasma for severe acute respiratory syndrome among healthcare workers in a Taiwan hospital

text: Objectives: To describe the immunological responses and clinical outcome of coronavirus (SARS) infected healthcare workers (HCW) who had been administered with convalescent plasma as a treatment. Methods: Convalescent plasma (500 mL) was obtained from each of three SARS patients and transfused into the three infected HCW. Donors were blood type O and seronegative for hepatitis B and C, HIV, syphilis and human T-cell lymphotropic virus types I and II (HTLV-I and -II). Serum antibody (IgG) titre was >640. Apharesis was performed with a CS 3000 plus cell separator followed by the forming of the convalescent phase plasma. As part of the routine check with donated plasma, the convalescent plasma was confirmed free of residual SARS-CoV by RT–PCR. Serial serum samples obtained from the recipients of the convalescent plasma were collected to undertake real-time quantitative RT–PCR for SARS-CoV for direct measurement of viral concentration. Specific immunoglobulin IgM and IgG concentrations were titrated using an antigen microarray developed in-house. Results: Viral load dropped from 495 × 10(3), 76 × 10(3) or 650 × 10(3) copies/mL to zero or 1 copy/mL one day after transfusion. Anti-SARS-CoV IgM and IgG also increased in a time-dependent manner following transfusion. All three patients survived. One HCW became pregnant subsequently, delivering 13 months after discharge. Positive anti-SARS-CoV IgG was detected in the newborn. Passive transfer of anti-SARS-CoV antibody from the mother was considered as a possibility. Conclusions: All infected HCW whose condition had progressed severely and who had failed to respond to the available treatment, survived after transfusion with convalescent plasma.


Introduction to molecular virology


title: Segmentation of DNA using simple recurrent neural network

text: Abstract We report the discovery of strong correlations between protein coding regions and the prediction errors when using the simple recurrent network to segment genome sequences. We are going to use SARS genome to demonstrate how we conduct training and derive corresponding results. The distribution of prediction error indicates how the underlying hidden regularity of the genome sequences and the results are consistent with the finding of biologists: predicated protein coding features of SARS genome. This implies that the simple recurrent network is capable of providing new features for further biological studies when applied on genome studies. The HA gene of influenza A subtype H1N1 is also analyzed in a similar way.


title: Risk of Waterborne Illness Via Drinking Water in the United States

text: The quality of drinking water in the United States is among the best in the world; however, waterborne disease outbreaks continue to occur, and many more cases of endemic illness are estimated. Documented waterborne disease outbreaks are primarily the result of technological failures or failure to treat the water (Craun et al. 2006). Current federal regulations require that all surface waters used for a drinking water supply be treated to reduce the level of pathogens so as to reduce the risk of infection to 1:10,000 per year (Regli et al. 1991). To achieve this goal, water treatment must, at a minimum, reduce infectious viruses by 99.99% and protozoan parasites by 99.9% (Regli et al. 2003). If Cryptosporidium concentrations exceed a certain level in the source water, additional reductions are required. This degree of treatment is usually achieved by a combination of physical processes (coagulation, sedimentation, and filtration) and disinfection (chlorination, ozonation). Filtration is essential for the removal of protozoan parasites due to their resistance to chlorination and ozonation at doses normally used in drinking water treatment (Barbeau et al. 2000; Korich et al. 1990; Rennecker et al. 1999). A variance from filtration is allowed in some cases if the watershed is protected and carefully monitored for protozoan pathogens.


title: EEG findings in acutely ill patients investigated for SARS‐CoV‐2/COVID‐19: A small case series preliminary report

text: OBJECTIVE: Acute encephalopathy may occur in COVID‐19‐infected patients. We investigated whether medically indicated EEGs performed in acutely ill patients under investigation (PUIs) for COVID‐19 report epileptiform abnormalities and whether these are more prevalent in COVID‐19 positive than negative patients. METHODS: In this retrospective case series, adult COVID‐19 inpatient PUIs underwent EEGs for acute encephalopathy and/or seizure‐like events. PUIs had 8‐channel headband EEGs (Ceribell; 20 COVID‐19 positive, 6 COVID‐19 negative); 2 more COVID‐19 patients had routine EEGs. Overall, 26 Ceribell EEGs, 4 routine and 7 continuous EEG studies were reviewed. EEGs were interpreted by board‐certified clinical neurophysiologists (n = 16). EEG findings were correlated with demographic data, clinical presentation and history, and medication usage. Fisher's exact test was used. RESULTS: We included 28 COVID‐19 PUIs (30‐83 years old), of whom 22 tested positive (63.6% males) and 6 tested negative (33.3% male). The most common indications for EEG, among COVID‐19‐positive vs COVID‐19‐negative patients, respectively, were new onset encephalopathy (68.2% vs 33.3%) and seizure‐like events (14/22, 63.6%; 2/6, 33.3%), even among patients without prior history of seizures (11/17, 64.7%; 2/6, 33.3%). Sporadic epileptiform discharges (EDs) were present in 40.9% of COVID‐19‐positive and 16.7% of COVID‐19‐negative patients; frontal sharp waves were reported in 8/9 (88.9%) of COVID‐19‐positive patients with EDs and in 1/1 of COVID‐19‐negative patient with EDs. No electrographic seizures were captured, but 19/22 COVID‐19‐positive and 6/6 COVID‐19‐negative patients were given antiseizure medications and/or sedatives before the EEG. SIGNIFICANCE: This is the first preliminary report of EDs in the EEG of acutely ill COVID‐19‐positive patients with encephalopathy or suspected clinical seizures. EDs are relatively common in this cohort and typically appear as frontal sharp waves. Further studies are needed to confirm these findings and evaluate the potential direct or indirect effects of COVID‐19 on activating epileptic activity.


title: High rates of 30-day mortality in patients with cirrhosis and COVID-19

text: BACKGROUND AND AIMS: Coronavirus disease (COVID-19) is a major worldwide threat for healthy individuals as well as for patients with comorbidities, but its impact on patients with cirrhosis is currently unknown. This study aimed at evaluating the impact of COVID-19 on the clinical outcome of these patients. METHODS: In this multicenter retrospective study, cirrhotic patients with confirmed Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) infection were enrolled between 1(st) and 31(th) March 2020. Clinical and biochemical data at COVID-19 and at the last outpatient visit were obtained through review of medical records. RESULTS: Fifty cirrhotic patients with confirmed SARS-CoV-2 infection were enrolled (age 67 years, 70% men, 38% virus-related, 52% previously compensated cirrhosis). At diagnosis, 64% of patients presented fever, 42% shortness of breath/polypnea, 22% encephalopathy, 96% needed hospitalization or prolonged an ongoing one. Respiratory-support was necessary in 71%, 52% received antivirals, 80% heparin. Serum albumin significantly decreased, while bilirubin, creatinine and prothrombin time significantly increased at COVID-19 diagnosis compared to last available data. The proportion of patients with MELD≥15 increased from 13% to 26% (p=0.037), acute-on-chronic liver failure and and de novo acute liver injury occurred in 14 (28%) and 10 patients, respectively. Seventeen patients died after a median of 10 (4-13) days from COVID-19 diagnosis, with a 30-day-mortality rate of 34%. Severity of lung and liver (according to CLIF-C, CLIF-OF and MELD scores) diseases independently predicted mortality. Mortality was significantly higher in hospitalized cirrhotics with COVID-19 than in those hospitalized for bacterial infections. CONCLUSION: COVID-19 is associated with liver function deterioration and elevated mortality in cirrhotic patients.


title: The microbiological characteristics of lower respiratory tract infection in patients with neuromuscular disorders: An investigation based on a multiplex polymerase chain reaction to detect viruses and a clone library analysis of the bacterial 16S rRNA gene sequence in sputum samples

text: Abstract We performed gene amplification methods for the detections of bacteria and viruses using sputum samples to clarify the microbiological characteristics of lower respiratory tract infection in patients with neuromuscular disorders. The tendencies of higher proportion of respiratory virus detection and lower diversity of bacteria in sputum were observed.


Are We There Yet? Laboratory Preparedness for Emerging Infectious Diseases


title: Neue Influenza A (H1N1) 2009

text: Abstract In April 2009 a novel influenza virus A H1N1 causing high morbidity and mortality spread first on the plateau of Mexico City subsequently followed by distribution to Texas and California in USA and to Spain; finally within three weeks this influenza virus was worldwide distributed. Its haemagglutinin showed epitope structures that were not recognized by the immune system of children and young adults as a memory response. The pandemic influenza virus consisted of quasispecies with a mixture of genome parts from human, avian, and American and Eurasian swine origin. This new virus caused all symptoms typically for influenza as abrupt onset, fever >39°C, sore throat with all signs of inflammation, myalgia and arthralgia. In summer 2009 all infections in Europe were mild, while until winter there was a higher lethality in Brazil, Mexico and Argentine; but still in winter 2009 in Europe the pathogenic action of the circulating virus was mild. European isolates showed a neuraminidase inhibitor resistance of around 10%. Prevention of spread of the novel influenza virus H1N1 2009 is mainly by hygienic measurements as: avoid contact to infected, distance from infected, hand disinfection, protection of eye and nose mucous membranes, avoid aerosol formation during blowing the nose and sneezing. It is recommended to use single way tissue handkerchiefs and to depose them in a covered waste reservoir. For further prevention of influenza virus transmission three different vaccines are available in Europe since October 2009. This virus or at least some genomic parts will persist in the human population for the next 2 years.


title: Evolution of virulence

text: At the close of the 19th century, the germ theory had generated a new understanding of the causes of acute infectious diseases and revealed new directions for study. This understanding contributed to the greatest improvements in health in the history of medicine. At the end of the 20th century, the second stage of this disciplinary development is occurring. The old germ theory is being expanded into a new germ theory, which, by integrated the full spectrum of biologic disciplines. This new germ theory is emphasizing how environments and human activities influence the characteristics of infectious agents and the broader role of infection as a cause of chronic diseases.


title: The EBV action in tonsils and adenoids

text: Abstract The bacteria involved in tonsil disease have been well studied, but we cannot say the same for the viruses. The method to detect virus make this approach difficult to study. Epstein–Barr Virus (EBV) infection usually occurs in early childhood and can persist in palatine and pharyngeal tonsil lymphocytes. EBV has been closely associated with the undifferentiated form of nasopharyngeal carcinoma (NPC) in its effect. Nevertheless, the presence of EBV in non-neoplastic lymphoid tissue of the nasopharynx and tonsil has rarely been investigated. Our objective was to study the frequency of EBV in tonsils and adenoids and to define the correlation between EBV and adenoid hyperplasia. In this study, we looked for EBV in adenoid and tonsil tissue of 165 patients (2 and 15 years old ) by in situ hybridization (ISH) for EBER 1/2 RNA. Resection of the adenoids was done for relief of upper respiratory tract obstruction, and the tonsils were resected because of recurrent tonsillitis and/or hyperplasia with upper airway obstruction. We divided the adenoid samples in two groups: one group 12–24 months old (average 18 months old) and the second group, 25 months to 15 years old. Tonsils were obtained from 85 patients, 3–13 years old (mean age 5.6 years) who underwent surgery due to recurrent tonsillitis or hyperplasia. EBV was demonstrated in lymphoid cells of 11 (34.3%) out of 32 adenoids for the first group and 36 (72%) out of 48 children of the second group. EBV was found in the respiratory epithelial cells of adenoid in one case. Children under 24 months of age can be infected by EBV, and this virus might be responsible for obstructive hyperplasia. Tonsils are less affected by EBV than the adenoids, suggesting that the EBV is more attracted to the adenoid tissue than the tonsillar tissue.


title: Fine mapping and conservation analysis of linear B-cell epitopes of peste des petits ruminants virus hemagglutinin protein

text: Abstract Hemagglutinin protein (H), one of the two glycoproteins of peste des petits ruminants virus (PPRV), binds to its receptor on the host cell and acts as a major antigen that induces and confers highly protective immunity in the host. In order to delineate the epitopes on H protein, fine epitope mapping and conservation analysis of linear B-cell epitopes (BCEs) on PPRV H has been undertaken using biosynthetic peptides and rabbit anti-PPRV H sera. Thirteen linear BCEs were identified and their corresponding minimal motifs were located on the H protein of PPRV China/Tibet/Geg/07-30. Conservation analysis indicated that two of the 13 minimal motifs were conserved among 52 PPRV strains. Nine of the 13 peptides containing the minimal motifs were recognized using anti-PPRV serum from a goat immunized with PPRV vaccine strain Nigeria 75/1. Identified epitopes and their motifs improve our understanding of the antigenic characteristics of PPRV H and provide a basis for the development of epitope-based diagnostic assays and multiple epitopes vaccine.


VASCULAR SURGERY ACTIVITY CONDITION IS A COMMON LANGUAGE FOR UNCOMMON TIMES


title: Non Pharmaceutical Interventions for Optimal Control of COVID-19

text: Background and Objective The outbreak of the current pandemic begun from the first individual of a 55-year old from Hubei province in China, the disease instigated by the new coronavirus spreading across the world. Scientists presently speculate this coronavirus, SARS-CoV-2, originated in a bat and by one way or another jumped to another creature, potentially the pangolin, which at that point gave it to people. The ailment is currently spreading between individuals with no animal delegate. Researchers are struggling to follow the infection back to where it started to become familiar with its spread. In the event that, for example, specialists can locate the soonest cases, they might have the option to distinguish the creature have where the infection hides. In March and April 2020, researchers detailed that this virus created normally. Coronavirus has been become of the serious global phenomena in the recent years and has negative effects in the entire world health and economy. The virus is believed to have been associated with a host animal which human contracted. Subsequently, human-to-human infection began. Through migration as humans have become complex with easy mobility the disease has traveled to the entire continent. Now, numerous scientist are going on in the hope of obtaining medication and vaccination to prevent the spread of the disease and mortality of the disease. It is important that we obtain quantitative and qualitative information about the etiology of this disease which is crucial. Mathematical modeling is capable of providing qualitative information on many parameters that guides the decision making of health practitioners. In this work we focus the optimal control of COVID-19 with the help of Non Pharmaceutical Interventions(NPIs). To find the role of factors/parameters in the transmission of the syndrome we find R(0); the ratio of reproduction for the proposed model. Methods To find the role of parameters in the transmission of the syndrome we find R(0); the ratio of reproduction for the proposed model. On the basis of sensitivity indices of the parameters we apply Non Pharmaceutical Interventions(NPIs) to control the sensitive parameters and hence formulate the optimal control mode. With the help of Hamiltonian and Lagrangian we minimize the density of contaminated stuff and infected human population. Results We focus the optimal control of COVID-19 with the help of Non Pharmaceutical Interventions(NPIs). On the basis of sensitivity indices of the parameters we apply Non Pharmaceutical Interventions(NPIs) to control the sensitive parameters and hence formulate the optimal control model. The major NPIs are, STAY HOME, SANITIZER (wash hands), EARLY CASE DETECTION (PCR Test) and FACE MASK. These NPIs helps in mitigation and reducing the size of outbreak of the disease. Conclusion We check the existence of the optimal solution for the system. At the end, Using matlab we produce numerical simulations for validation of results of control variables. The results demonstrate that if there is no control (variables/interventios), 900 out 1000 susceptible individuals may be infected (exposed) in very short period. As such a circumstances no agency fighting against COVID-19 could be successful due to its limited resources. Keywords: Novel coronavirus, Mathematical model, Basic reproduction number, Next generation matrix, Sensitivity analysis, Pontryagin’s Maximum Principle, Optimal control.


title: Der Ausbruch von COVID-19 in China

text: The transmission dynamics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Wuhan and Hubei Province differ considerably from those in the rest of China. In Hubei province SARS-CoV‑2 led to a dramatic outbreak. Intensive control measures (travel restrictions, isolation of cases, quarantine of contacts and others) led to the control of the outbreak. Despite travel restrictions SARS-CoV‑2 was detected in other provinces in the following weeks. Consistent and intensive identification and isolation of infected persons (“containment”) was able to prevent an outbreak outside Hubei province, providing an example for the control of SARS-CoV‑2.


title: Myelin basic protein and human coronavirus 229E cross‐reactive T cells in multiple sclerosis

text: Multiple sclerosis (MS) is an inflammatory demyelinating neurological disease in which autoreactive T lymphocytes sensitized to myelin components of the central nervous system are postulated to contribute to pathogenesis. The possible relevance of molecular mimicry between a human coronavirus and the myelin basic protein component of myelin in the generation of this autoimmune reaction was evaluated. Myelin basic protein– and virus‐reactive T‐cell lines were established from 16 MS patients and 14 healthy donors and shown to be mostly CD4(+). In contrast to healthy donors, several T‐cell lines isolated from MS patients showed cross‐reactivity between myelin and coronavirus antigens. Overall, 29% of T‐cell lines from MS patients (10 donors) but only 1.3% of T‐cell lines from normal control subjects (2 donors) showed an HLA‐DR‐restricted cross‐reactive pattern of antigen activation after in vitro selection with either myelin basic protein or human coronavirus strain 229E antigens. Moreover, reciprocal reactivities were only observed in MS patients (4 donors). This establishes molecular mimicry between a common viral pathogen, such as this human coronavirus, and myelin as a possible immunopathological mechanism in MS and is consistent with the possible involvement of more than one infectious pathogen as an environmental trigger of disease.


title: SARS-CoV-2 infection and stem cells: interaction and intervention

text: The emergence of the novel severe acute respiratory coronavirus 2 (SARS-CoV-2) in China and its rapid national and international spread have created a global health emergency. The resemblance with SARS-CoV in spike protein suggests that SARS-CoV-2 employs spike–driven entry into angiotensin-converting enzyme 2 (ACE2)-expressing cells. From a stem cell perspective, this review focuses on the possible involvement of ACE2(+) stem/progenitor cells from both the upper and lower respiratory tracts in coronavirus infection. Viral infection-associated acute respiratory distress syndrome and acute lung injury occur because of dysregulation of the immune response. Mesenchymal stem cells appear to be a promising cell therapy given that they favorably modulate the immune response to reduce lung injury. The use of exogenous stem cells may lead to lung repair. Therefore, intervention by transplantation of exogenous stem cells may be required to replace, repair, remodel, and regenerate lung tissue in survivors infected with coronavirus. Ultimately, vaccines, natural killer cells and induced-pluripotent stem cell-derived virus-specific cytotoxic T lymphocytes may offer off-the-shelf therapeutics for preventing coronavirus reemergence.


title: Characterization of the nuclear and nucleolar localization signals of bovine herpesvirus-1 infected cell protein 27

text: Abstract Bovine herpesvirus-1 infected cell protein 27 (BICP27) was detected predominantly in the nucleolus. The open reading frame of BICP27 was fused with the enhanced yellow fluorescent protein (EYFP) gene to investigate its subcellular localization in live cells and BICP27 was able to direct monomeric, dimeric or trimeric EYFP exclusively to the nucleolus. By constructing a series of deletion mutants, the putative nuclear localization signal (NLS) and nucleolar localization signal (NoLS) were mapped to 81RRAR84 and 86RPRRPRRRPRRR97 respectively. Specific deletion of the putative NLS, NoLS or both abrogated nuclear localization, nucleolar localization or both respectively. Furthermore, NLS was able to direct trimeric EYFP predominantly to the nucleus but excluded from the nucleolus, whereas NoLS targeted trimeric EYFP primarily to the nucleus, and enriched in the nucleolus with faint staining in the cytoplasm. NLS+NoLS directed trimeric EYFP predominantly to the nucleolus with faint staining in the nucleus. Moreover, deletion of NLS+NoLS abolished the transactivating activity of BICP27 on gC promoter, whereas deletion of either NLS or NoLS did not. The study demonstrated that BICP27 is a nucleolar protein, adding BICP27 to the growing list of transactivators which localize to the nucleolus.


Coronavirus Outbreak Takes Its Toll on Global Automotive Industry


title: Personal Protective Equipment and Antiviral Drug Use during Hospitalization for Suspected Avian or Pandemic Influenza(1)

text: For pandemic influenza planning, realistic estimates of personal protective equipment (PPE) and antiviral medication required for hospital healthcare workers (HCWs) are vital. In this simulation study, a patient with suspected avian or pandemic influenza (API) sought treatment at 9 Australian hospital emergency departments where patient–staff interactions during the first 6 hours of hospitalization were observed. Based on World Health Organization definitions and guidelines, the mean number of “close contacts” of the API patient was 12.3 (range 6–17; 85% HCWs); mean “exposures” were 19.3 (range 15–26). Overall, 20–25 PPE sets were required per patient, with variable HCW compliance for wearing these items (93% N95 masks, 77% gowns, 83% gloves, and 73% eye protection). Up to 41% of HCW close contacts would have qualified for postexposure antiviral prophylaxis. These data indicate that many current national stockpiles of PPE and antiviral medication are likely inadequate for a pandemic.


title: Changes in cecal microbiota community of suckling piglets infected with porcine epidemic diarrhea virus

text: Diarrhea, caused by porcine epidemic diarrhea virus (PEDV), is a catastrophic gastrointestinal disease among suckling piglets, with high infectivity, morbidity, and mortality, causing huge economic losses to the pig industry. In the present study, we investigated the different microbiota from the cecal mucosa and cecal contents between healthy and PEDV-infected piglets. High-throughput 16S rRNA gene sequencing was performed to explore differences. The results revealed that microbial dysbiosis by PEDV infection occurred in the cecal mucosa and contents of suckling piglets at each microbial taxonomic level. The abundance of pathogenic bacteria associated with diseases, including diarrhea, was increased. The abundance of Fusobacterium was 26.71% and 33.91% in cecal mucosa and contents of PEDV-infected group, respectively, whereas that in the healthy groups was 17.85% and 9.88%. The proportion of Proteobacteria in the infected groups was relatively high (24.67% and 22.79%, respectively), whereas that in the healthy group was 13.13% and 11.34% in the cecal mucosa and contents, respectively. Additionally, the proportion of Bacteroidetes in the healthy group (29.89%, 37.32%) was approximately twice that of the PEDV-infected group (15.50%, 15.39%). “Nitrate reduction”, “Human pathogens diarrhea”, “Human pathogens gastroenteritis”, “Nitrite respiration”, and “Nitrite ammonification” were the enriched functional annotation terms in the PEDV-infected groups. Porcine epidemic diarrhea virus infection increased the proportion of harmful bacteria and decreased the proportion of beneficial bacteria in the cecal mucosa and contents of suckling piglets. Our findings suggest that determining the intestinal microbiota might provide a promising method to prevent PEDV and open a new avenue for future research.


title: Design, synthesis, and biological evaluation of novel dipeptide-type SARS-CoV 3CL protease inhibitors: Structure–activity relationship study

text: This work describes the design, synthesis, and evaluation of low-molecular weight peptidic SARS-CoV 3CL protease inhibitors. The inhibitors were designed based on the potent tripeptidic Z-Val-Leu-Ala(pyrrolidone-3-yl)-2-benzothiazole (8; K(i) = 4.1 nM), in which the P3 valine unit was substituted with a variety of distinct moieties. The resulting series of dipeptide-type inhibitors displayed moderate to good inhibitory activities against 3CL(pro). In particular, compounds 26m and 26n exhibited good inhibitory activities with K(i) values of 0.39 and 0.33 μM, respectively. These low-molecular weight compounds are attractive leads for the further development of potent peptidomimetic inhibitors with pharmaceutical profiles. Docking studies were performed to model the binding interaction of the compound 26m with the SARS-CoV 3CL protease. The preliminary SAR study of the peptidomimetic compounds with potent inhibitory activities revealed several structural features that boosted the inhibitory activity: (i) a benzothiazole warhead at the S1′ position, (ii) a γ-lactam unit at the S1-position, (iii) an appropriately hydrophobic leucine moiety at the S2-position, and (iv) a hydrogen bond between the N-arylglycine unit and a backbone hydrogen bond donor at the S3-position.


title: COVID-19 is likely to impact animal health

text: Responses to the ongoing COVID-19 pandemic have included travel bans and social distancing with “shelter in place” orders, resulting in sudden changes in human activity and subsequent effects on the global and national economy. We speculate that animal health will likely be impacted by COVID-19 through the immediate consequences of sudden human confinement and inactivity, and through the long-term consequences of the upcoming economic crisis on farmer livelihoods and veterinary service capacities. We expect the COVID-19 pandemic and the subsequent economic crisis to impact negatively on the control of diseases that are already present in Europe, as well as on the European capacity to prevent and respond in a timely manner to new and emerging animal diseases. We also expect an increased attention to the animal health implications of coronavirus infections in animals. Mechanisms explaining these outcomes include increased wildlife-livestock contacts due to human confinement; disruption of ongoing testing schemes for endemic diseases; lower disease surveillance efforts; and lower capacity for managing populations of relevant wildlife reservoirs. The main mitigation action consists in adapting animal health management strategies to the available resources.


SARS-CoV-2 related liver impairment – perception may not be the reality


title: Development of antiviral therapy for severe acute respiratory syndrome

text: Abstract A new disease, the severe acute respiratory distress syndrome (SARS), caused by the SARS coronavirus (SARS-CoV), emerged at the beginning of 2003 and rapidly spread throughout the world. Although the disease had disappeared in June 2003 its re-emergence cannot be excluded. The development of vaccines against SARS-CoV may take years. Therefore, the availability of effective antiviral drugs against SARS-CoV may be crucial for the control of future SARS outbreaks. In this review, experimental and clinical data about potential anti-SARS drugs is summarised and discussed. Animal model studies will be needed to help to determine which interventions warrant controlled clinical testing.


title: Sequence and analysis of bovine enteritic coronavirus (F15) genome I.—Sequence of the gene coding for the nucleocapsid protein; analysis of the predicted protein

text: Summary Sequences encoding the N protein of the bovine enteritic coronavirus-F15 strain (BECV-F15) have been cloned in PBR322 plasmid using cDNA produced by priming with oligo-dT on purified viral genomic RNA. Some 265 insert-containing clones were studied. Hybridization of these inserts with poly(A)+ RNA extracted from infected cells led to the conclusion that they were located at the 3′-end of the genome. After subcloning in M13 phage DNA, clones were sequenced by the Sanger technique. A 1,710-nucleotide sequence corresponding to the gene coding for the viral N-protein was established. It shows 2 overlapping open reading frames (ORF). The 3′-non-coding end of the gene has an 8-nucleotide sequence in common with the homologous genome areas of MHV, TGE and IBV viruses. This sequence may represent the polymerase RNA binding site. An upstream sequence surrounding the first AUG of the smaller ORF corresponds to a potentially functional initiation codon. The sequence of the primary translation product deduced from the DNA sequence predicts a polypeptide of 207 amino acids (22.9 Kd) with a high leucine (19.8%) content, possessing a hydrophobic N-terminal end. The larger ORF has a coding capacity of 448 amino acids (49.4 Kd), corresponding to the N-protein molecular weight. The deduced protein possesses 43 serine residues (9.6% of the total amino acid content) which may be phosporylated and involved in N-protein/RNA binding. N-protein also has 5 regions with a high basic amino acid content. One of them is also serine-rich and has a strong homology site with MHV, TGE and IBV viruses. In the first part of the N-terminal, a 12-amino-acid sequence (PRWYFYYLGTGP) is highly conserved for BECV-F15, JHM, TGE and IBV viruses. BCV Mebus strain and BECV-F15 have only minor differences in their N-protein sequence.


title: Inhaled corticosteroid use is associated with increased circulating T regulatory cells in children with asthma

text: BACKGROUND: T regulatory (Treg) cells are important in balancing immune responses and dysregulation of Treg cells has been implicated in the pathogenesis of multiple disease states including asthma. In this study, our primary aim was to determine Treg cell frequency in the peripheral blood of children with and without asthma. The secondary aim was to explore the association between Treg cell frequency with allergen sensitization, disease severity and medication use. METHODS: Peripheral blood mononuclear cells from healthy control subjects (N = 93) and asthmatic children of varying disease severity (N = 66) were characterized by multi-parameter flow cytometry. RESULTS: Our findings demonstrate that children with asthma had a significantly increased frequency of Treg cells compared to children without asthma. Using a multivariate model, increased Treg cell frequency in children with asthma was most directly associated with inhaled corticosteroid use, and not asthma severity, allergic sensitization, or atopic status of the asthma. CONCLUSION: We conclude that low dose, local airway administration of corticosteroids is sufficient to impact the frequency of Treg cells in the peripheral blood. These data highlight the importance of considering medication exposure when studying Treg cells and suggest inhaled corticosteroid use in asthmatics may improve disease control through increased Treg cell frequency.


title: Pan-human coronavirus and human bocavirus SYBR Green and TaqMan PCR assays; use in studying influenza A viruses co-infection and risk of hospitalization

text: PURPOSE: Influenza A viruses, human coronaviruses (hCoV) and human bocavirus (hBoV) are emerging respiratory viruses. This study investigated the association between influenza A viruses co-infection with hBoV and hCoV and severity and the sensitivity of a real-time polymerase chain reaction (RT-PCR) assay for identification of 15 coronaviruses. METHODOLOGY: Published sequences for the 15 human coronaviruses were used to design a consensus PCR targeting the replicase open reading frame 1b. A previously published PCR targeting the NS1 Gene of all known human bocavirus strains was also utilized. A series of 217 samples from patients aged 37.7 (SD ± 30.4)] with seasonal influenza A viruses (SeasFluA) identified between 06/2011 and 06/2012 in NW England were tested for hCoV and hBoV using RT-PCR. Association between co-infection and disease outcome was assessed using logistic regression. RESULTS: The limit of detection of hCoV RT-PCR assay was 2 copies/µl of human coronavirus RNA template, a sensitivity comparable to a previously published SYBR green assay for human coronaviruses. A total of 12 hCoV and 17 hBoV were identified in the 217 influenza A positive samples. A higher proportion (61.5 %; 8/13) of SeasFluA/hBoV co-infections were identified in patients that were admitted either to a general ward or the intensive care unit compared to 44.3 % (66/149) of single SeasFlu A virus infections (OR 2.5 95 % CI 0.67–9.34, p = 0.17). In a stratified analysis, there was a trend towards higher association between FluA, hCoV and hBoV with increasing age (especially in patients aged 24–45 years and >65 year old). CONCLUSION: Our hCoV RT-PCR protocol appeared to be of adequate analytical sensitivity for diagnosis. More and larger studies are needed to confirm the role of hCoV, hBoV in causing severe disease when they co-infect with influenza A viruses.


title: Redundancy in reporting on COVID‐19

text: Additional to the problems described very well by Dr. Ioannidis1 , there is another issue that became highlighted during this pandemic: redundancy in research and reporting. Redundant articles repeat already known information and are mostly published just for the sake of publishing. By searching through Pubmed, one can easily see that the number of publications related to COVID-19 is growing exponentially (Figure).


Sense and sensibility through confusing surgical practices during COVID‐19 pandemic


Is the type of diabetes treatment relevant to outcome of COVID‐19?


title: Widespread distribution and a new recombinant species of Brazilian virus associated with cotton blue disease

text: BACKGROUND: Cotton blue disease (CBD), an important global cotton crop pathology responsible for major economic losses, is prevalent in the major cotton-producing states of Brazil. Typical CBD symptoms include stunting due to internodal shortening, leaf rolling, intense green foliage, and yellowing veins. Atypical CBD symptoms, including reddish and withered leaves, were also observed in Brazilian cotton fields in 2007. Recently, a Polerovirus named Cotton leafroll dwarf virus (CLRDV) was shown to be associated with CBD. RESULTS: To understand the distribution and genetic diversity of CLRDV in Brazil, we analyzed 23 CBD-symptomatic plants from susceptible cotton varieties originating from five of the six most important cotton-growing states, from 2004–2007. Here, we report on CLRDV diversity in plants with typical or atypical CBD symptoms by comparing viral coat protein, RNA polymerase (RdRp), and intergenic region genomic sequences. CONCLUSION: The virus had a widespread distribution with a low genetic diversity; however, three divergent isolates were associated with atypical CBD symptoms. These divergent isolates had a CLRDV-related coat protein but a distinct RdRp sequence, and probably arose from recombination events. Based on the taxonomic rules for the family Luteoviridae, we propose that these three isolates represent isolates of a new species in the genus Polerovirus.


title: Comparative Serological Study for the Prevalence of Anti-MERS Coronavirus Antibodies in High- and Low-Risk Groups in Qatar

text: Infection with Middle East respiratory syndrome coronavirus (MERS-CoV) could be asymptomatic or cause mild influenza-like illness. Therefore, the prevalence of MERS-CoV infections in the general population could be underestimated, which necessitates active surveillance to determine the epidemiological importance of asymptomatic cases. The aim of this study is to evaluate the performance of various serological assays and to estimate the seroprevalence of anti-MERS-CoV antibodies in high- and low-risk groups in Qatar. A total of 4858 samples were screened, including 4719 samples collected from healthy blood donors (BD) over a period of five years (2012-2016), 135 samples from baseline case contacts (CC) collected from individuals in close contact with three positive PCR-confirmed patients (CP), and four samples from MERS-CoV CP. Initial screening using anti-MERS-CoV IgG (IgG rS1-ELISA kit) revealed ten reactive samples from BD (10/4719, 0.21%), one from CC (1/135, 0.74%), and three from CP (3/4, 75%). Samples from CP but not from BD were also reactive by whole-virus anti-MERS-CoV IgG (n = 3/4) and IgM (n = 1/4) indirect immunefluorescent tests (IIFT) and pseudoparticle neutralization test (ppNT). The reactive sample from CC was also confirmed by ppNT. Surprisingly, one out of thirteen (7.7%) randomly selected IgG rS1-ELISA-negative BD samples from the initial screening was reactive by the IgM-IIFT (but not by the IgG-IIFT) and was subsequently confirmed by ppNT. All IgG rS1-ELISA-reactive samples from BD exhibited considerable reactivity to the four circulating human coronaviruses (HKU1, OC43, 229E, and NL63). Cross-reactivity with SARS was only reported for samples from CP using IgG and IgM-IIFT. In conclusion, we report a low prevalence of anti-MERS antibodies in the general population, which coincides with the low number of all reported cases by the time of our study (2017) in Qatar (n = 21). The false-positive results and the observed cross-reactivity between MERS-CoV and other circulating human coronavirus necessitate more detailed evaluation of available serological assays.


title: Assessment of intervention strategies against a novel influenza epidemic using an individual-based model

text: OBJECTIVES: The objective of this study was to assess intervention strategies against a novel influenza epidemic through simulations of various scenarios in Sapporo city, Hokkaido, Japan. A series of interventions were examined: administration of antiviral drugs by two approaches [targeted antiviral prophylaxis (TAP) and school-age targeted antiviral prophylaxis (STAP)], school closure, restraint, and combinations of these four interventions. METHODS: In order to generate a more realistic situation, we constructed an individual-based model (IBM) for the transmission of influenza in which each individual was assigned personal information on the basis of the National Census and Employment Status Survey of Sapporo city. In addition, data on high-risk casual contact groups commuting in crowded trains and buses were obtained from a census on transportation modes and introduced into the model. Observational data from previous pandemics were used for the epidemiological parameters. RESULTS: Both TAP and STAP interventions were highly effective in suppressing the spread of infection during the early period of an outbreak, but STAP was inferior to TAP in terms of the ripple effect of the administration of antiviral drugs. School closure and restraint were able to bring about a delay in the peak of infection. The combination of TAP, school closure, and restraint interventions were highly effective in decreasing the total number of patients and shortening the epidemic period. CONCLUSIONS: Based on the simulation results, we recommend implementing TAP together with both school closure and restraint as strategies against a future novel influenza outbreak.


Co‐occurrence of birds and bats in natural nest‐holes


title: Silver nanoparticle-doped zirconia capillaries for enhanced bacterial filtration

text: Abstract Membrane clogging and biofilm formation are the most serious problems during water filtration. Silver nanoparticle (Agnano) coatings on filtration membranes can prevent bacterial adhesion and the initiation of biofilm formation. In this study, Agnano are immobilized via direct reduction on porous zirconia capillary membranes to generate a nanocomposite material combining the advantages of ceramics being chemically, thermally and mechanically stable with nanosilver, an efficient broadband bactericide for water decontamination. The filtration of bacterial suspensions of the fecal contaminant Escherichia coli reveals highly efficient bacterial retention capacities of the capillaries of 8 log reduction values, fulfilling the requirements on safe drinking water according to the U.S. Environmental Protection Agency. Maximum bacterial loading capacities of the capillary membranes are determined to be 3×109 bacterialcells/750mm2 capillary surface until back flushing is recommendable. The immobilized Agnano remain accessible and exhibit strong bactericidal properties by killing retained bacteria up to maximum bacterial loads of 6×108 bacterialcells/750mm2 capillary surface and the regenerated membranes regain filtration efficiencies of 95–100%. Silver release is moderate as only 0.8% of the initial silver loading is leached during a three-day filtration experiment leading to average silver contaminant levels of 100μg/L.


Addressing Skilled Nursing Facilities' COVID-19 Psychosocial Needs via Staff Training and a Process Group Intervention


title: Cholesterol enhances mouse hepatitis virus-mediated cell fusion

text: Abstract Mouse hepatitis virus (MHV) infection of the L-2 subline of mouse fibroblasts results in acute infection characterized by extensive cell fusion. In contrast, infection of the LM-K subline leads to virus persistence with reduced cell fusion. We undertook studies designed to elucidate the role of host cell membrane lipid composition and the cytoskeleton in modulating the fusion process and the resultant effect(s) on virus persistence. MHV-induced cell fusion proceeded normally in cells treated with cytoskeleton-disrupting drugs, cytochalasin B and colchicine. Modification of cell membrane fatty acid composition by supplementation of LM-K cells with arachidonic (C-20:4) or palmitic (C-16:0) acids had little effect on the extent of MHV-induced cell fusion or on virus replication. However, supplementation of both cell types with cholesterol (resulting in increased membrane cholesterol/fatty acid ratio) resulted in marked enhancement of virus-mediated cell fusion. The increase in cell membrane cholesterol did not enhance internalization of MHV suggesting that cholesterol primarily modulates a later event. This suggestion was confirmed by demonstrating cholesterol-enhancement of fusion in a contact fusion assay. Cholesterol-supplemented L-2 cells were less productive for virus replication than unsupplemented cells, in agreement with our previous observations that MHV replication is compromised by extensive cytopathic effect. Although cholesterol-supplemented LM-K cells showed increased susceptibility to MHV-mediated cell fusion, the extent of such susceptibility did not approach that observed in L-2 cells. Also, the property of LM-K cells to support MHV persistence was not abolished by cholesterol supplementation. Thus membrane fusion resistance and MHV persistence are modulated but not alleviated by cell membrane cholesterol content.


title: Phylogenetic analysis and sequence comparisons of structural and non-structural SARS coronavirus proteins in Taiwan

text: Abstract Taiwan experienced a large number of severe acute respiratory syndrome (SARS) viral infections between March and July 2003; by September of that year, 346 SARS cases were confirmed by RT-PCR or serological tests. In order to better understand evolutionary relationships among SARS coronaviruses (SCoVs) from different international regions, we performed phylogenetic comparisons of full-length genomic and protein sequences from 45 human SCoVs (including 12 from Taiwan) and two civet SCoVs. All the Taiwanese SARS-CoV strains which associated with nosocomial infection formed a monophyletic clade within the late phase of the SARS epidemic. This Taiwanese clade could be further divided into two epidemic waves. Taiwan SCoVs in the first wave clustered with three isolates from the Amoy Gardens housing complex in Hong Kong indicating their possible origin. Of the 45 human SCoVs, one isolate from Guangdong province, China, exhibited an extra 29-nucleotide fragment between Orf 10 and Orf 11—similar to the civet SCoV genome. Nucleotide and protein sequence comparisons suggested that all SCoVs of late epidemic came from human-to-human transmission, while certain SCoVs of early epidemic might have originated in animals.


title: Evaluation of the lockdowns for the SARS-CoV-2 epidemic in Italy and Spain after one month follow up

text: Abstract From the end of February, the SARS-CoV-2 epidemic in Spain has been following the footsteps of that in Italy very closely. We have analyzed the trends of incident cases, deaths, and intensive care unit admissions (ICU) in both countries before and after their respective national lockdowns using an interrupted time-series design. Data was analyzed with quasi-Poisson regression using an interaction model to estimate the change in trends. After the first lockdown, incidence trends were considerably reduced in both countries. However, although the slopes have been flattened for all outcomes, the trends kept rising. During the second lockdown, implementing more restrictive measures for mobility, it has been a change in the trend slopes for both countries in daily incident cases and ICUs. This improvement indicates that the efforts overtaken are being successful in flattening the epidemic curve, and reinforcing the belief that we must hold on.


title: Effects of personalized ventilation interventions on airborne infection risk and transmission between occupants

text: The role of personalized ventilation (PV) in protecting against airborne disease transmission between occupants was evaluated by considering two scenarios with different PV alignments. The possibility that PV may facilitate the transport of exhaled pathogens was explored by performing experiments with droplets and applying PV to a source or/and a target manikin. The risk of direct and indirect exposure to droplets in the inhalation zone of the target was estimated, with these exposure types defined according to their different origins. The infection risk of influenza A, a typical disease transmitted via air, was predicted based on a dose-response model. Results showed that the flow interactions between PV and the infectious exhaled flow would facilitate airborne transmission between occupants in two ways. First, application of PV to the source caused more than 90% of indirect exposure of the target. Second, entrainment of the PV jet directly from the infectious exhalation increased direct exposure of the target by more than 50%. Thus, these scenarios for different PV application modes indicated that continuous exposure to exhaled influenza A virus particles for 2 h would correspond with an infection probability ranging from 0.28 to 0.85. These results imply that PV may protect against infection only when it is maintained with a high ventilation efficiency at the inhalation zone, which can be realized by reduced entrainment of infectious flow and higher clean air volume. Improved PV design methods that could maximize the positive effects of PV on disease control in the human microenvironment are discussed.


Index to Subjects ∗ ∗ July, pp. 1-98; August, pp. 99-198; September, pp. 199-294; October, pp. 295-438; November, pp. 439-564; December, pp. 565-736.


title: Hepatitis C virus core protein binding to lipid membranes: the role of domains 1 and 2

text: Summary. We have analysed and identified different membrane‐active regions of the Hepatitis C virus (HCV) core protein by observing the effect of 18‐mer core‐derived peptide libraries from two HCV strains on the integrity of different membrane model systems. In addition, we have studied the secondary structure of specific membrane‐interacting peptides from the HCV core protein, both in aqueous solution and in the presence of model membrane systems. Our results show that the HCV core protein region comprising the C‐terminus of domain 1 and the N‐terminus of domain 2 seems to be the most active in membrane interaction, although a role in protein–protein interaction cannot be excluded. Significantly, the secondary structure of nearly all the assayed peptides changes in the presence of model membranes. These sequences most probably play a relevant part in the biological action of HCV in lipid interaction. Furthermore, these membranotropic regions could be envisaged as new possible targets, as inhibition of its interaction with the membrane could potentially lead to new vaccine strategies.


title: Melatonin is responsible for rice resistance to rice stripe virus infection through a nitric oxide-dependent pathway

text: Rice stripe virus (RSV) causes one of the most important rice virus diseases of plants in East Asia. However, the molecular mechanisms controlling rice resistance to RSV infection are largely unknown. Recently, several studies presented a novel model that melatonin (MT) and nitric oxide (NO) participate in the plant-pathogen interaction in a synergetic manner. In this study, there was a difference in MT content between two rice varieties that correlated with one being susceptible and one being resistant to RSV, which suggested that MT is related to RSV resistance. In addition, a test with two NO biosynthesis inhibitors revealed that NO inhibitor were able to increase the disease incidence of RSV. A pharmacological experiment with exogenous MT and NO showed that increased MT and NO in the MT-pretreated plants led to lower disease incidences; however, only NO increased in a NO-releasing reagent [sodium nitroprusside (SNP)] pretreated plants. The expressions level of OsPR1b and OsWRKY 45 were significantly induced by MT and NO. These results suggest that rice resistance to RSV can be improved by increased MT through a NO-dependent pathway.


title: Possible Bat Origin of Severe Acute Respiratory Syndrome Coronavirus 2

text: We showed that severe acute respiratory syndrome coronavirus 2 is probably a novel recombinant virus. Its genome is closest to that of severe acute respiratory syndrome–related coronaviruses from horseshoe bats, and its receptor-binding domain is closest to that of pangolin viruses. Its origin and direct ancestral viruses have not been identified.


title: The LXR ligand GW3965 inhibits Newcastle disease virus infection by affecting cholesterol homeostasis

text: Newcastle disease (ND) is a contagious disease that affects most species of birds. Its causative pathogen, Newcastle disease virus (NDV), also exhibits considerable oncolytic activity against mammalian cancers. A better understanding of the pathogenesis of NDV will help us design efficient vaccines and novel anticancer strategies. GW3965, a widely used synthetic ligand of liver X receptor (LXR), induces the expression of LXRs and its downstream genes, including ATP-binding cassette transporter A1 (ABCA1). ABCA1 regulates cellular cholesterol homeostasis. Here, we found that GW3965 inhibited NDV infection in DF-1 cells. It also inhibited NF-κB activation and reduced the upregulation of proinflammatory cytokines induced by the infection. Further studies showed that GW3965 exerted its inhibitory effects on virus entry and replication. NDV infection increased the mRNA levels of several lipogenic genes but decreased the ABCA1 mRNA level. Overexpression of ABCA1 inhibited NDV infection and reduced the cholesterol content in DF-1 cells, but when the cholesterol was replenished, NDV infection was restored. GW3965 treatment prevented cholesterol accumulation in the perinuclear area of the infected cells. In summary, our studies suggest that GW3965 inhibits NDV infection, probably by affecting cholesterol homeostasis.


title: How we treat patients with lung cancer during the SARS-CoV-2 pandemic: primum non nocere

text: New cases of the novel coronavirus, also known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to rise worldwide. A few reports have showed that mortality due to SARS-CoV-2 is higher in elderly patients and other active comorbidities including cancer. To date, no effective treatment has been identified and management for critically ill patients relies on management in intensive care units. Patients with lung cancer are at risk of pulmonary complications from COVID-19. Furthermore, the use of chemotherapy might have a negative impact in patient’s outcome. Therefore, the risk/benefit ratio of systemic anticancer treatment (SACT) has to be considered. For each patient, several factors including age and comorbidities, as well as the number of hospital visits for treatment, can influence this risk. Each hospital around the world has issued some internal policy guidelines for oncologists, aiming to limit risks during this difficult time. We hereby propose a tool to support oncologists and physicians in treatment decision for patients with lung cancer. There are several variables to consider, including the extent of the epidemic, the local healthcare structure capacity, the risk of infection to the individual, the status of cancer, patients’ comorbidities, age and details of the treatment. Given this heterogeneity, we have based our suggestions bearing in mind some general factors There is not easy, universal solution to oncological care during this crisis and, to complicate matters, the duration of this pandemic is hard to predict. It is important to weigh the impact of each of our decisions in these trying times rather than rely on routine automatisms.


Data Safety Monitoring during Covid‐19: Keep On Keeping On


Understanding the structural organization of lipids in the cell and viral membranes is essential for elucidating mechanisms of viral fusion that lead to entry of enveloped viruses into their host cells. The HIV lipidome shows a remarkable enrichment in dihydrosphingomyelin, an unusual sphingolipid formed by a dihydrosphingosine backbone. Here we investigated the ability of dihydrosphingosine to incorporate into the site of membrane fusion mediated by the HIV envelope (Env) protein. Dihydrosphingosine as well as cholesterol, fatty acid, and tocopherol was conjugated to highly conserved, short HIV-1 Env-derived peptides with no antiviral activity otherwise. We showed that dihydrosphingosine exclusively endowed nanomolar antiviral activity to the peptides (IC(50) as low as 120 nM) in HIV-1 infection on TZM-bl cells and on Jurkat T cells, as well as in the cell-cell fusion assay. These sphingopeptides were active against enfuvirtide-resistant and wild-type CXCR4 and CCR5 tropic HIV strains. The anti-HIV activity was determined by both the peptides and their dihydrosphingosine conjugate. Moreover, their mode of action involved accumulation in the cells and viruses and binding to membranes enriched in sphingomyelin and cholesterol. The data suggest that sphingopeptides are recruited to the HIV membrane fusion site and provide a general concept in developing inhibitors of sphingolipid-mediated biological systems.—Ashkenazi, A., Viard, M., Unger, L., Blumenthal, R., Shai, Y. Sphingopeptides: dihydrosphingosine-based fusion inhibitors against wild-type and enfuvirtide-resistant HIV-1.


title: The effect of population health on foreign direct investment inflows to low- and middle-income countries

text: Summary This paper investigates the effect of population health on gross inflows of foreign direct investment (FDI). We conduct a panel data analysis of 74 industrialized and developing countries over 1980–2000. Our main finding is that gross inflows of FDI are strongly and positively influenced by population health in low- and middle-income countries. Our estimates suggest that raising life expectancy by one year increases gross FDI inflows by 9%, after controlling for other relevant variables. These findings are consistent with the view that health is an integral component of human capital for developing countries.


title: “N95/N99” Peri-Procedure Diapers During Lower Gastrointestinal Endoscopies

text: It is important to consider lower gastrointestinal endoscopies (LGIE) as aerosol-generating procedures. Thus, it may be better to protect room environments by ensuring patients wearing peri-procedure diapers (PPD) to contain infectious colorectal gas expulsions because fecal SARS-CoV-2 has been detected among COVID-19 patients even after they have undetectable nasopharyngeal SARS-CoV-2. Summarily, PPD among LGIE patients can potentially evolve as standard barrier modality.


title: The role of phagocytic cells in enhanced susceptibility of broilers to colibacillosis after Infectious Bronchitis Virus infection

text: Abstract Colibacillosis results from infection with avian pathogenic Escherichia coli bacteria. Healthy broilers are resistant to inhaled E. coli, but previous infection with vaccine or virulent strains of Infectious Bronchitis Virus (IBV) predisposes birds for severe colibacillosis. We investigated whether IBV affects recruitment and function of phagocytic cells and examined NO production, phagocytic and bactericidal activity, and kinetics of peripheral blood mononuclear cells (PBMC) and splenocytes. Moreover, we measured cytokine mRNA expression in lung and spleen samples. Broilers were inoculated with IBV H120 vaccine or virulent M41 and challenged 5 days later with E. coli 506. A PBS control and E. coli group without previous virus inoculation were also included. Birds were sacrificed at various time points after inoculation (h/dpi). Inoculation with IBV induced extended and more severe colibacillosis than with E. coli alone. At 4dpi, the number of KUL-01+ PBMC in all E. coli-inoculated groups was significantly higher than in PBS-inoculated birds, which correlated with lesion scores. From 1 to 4dpi, NO production by PBMC from all E. coli-inoculated animals was elevated compared to PBS birds. Bactericidal activity of PBMC in IBV-inoculated animals at 7dpi was lower than in PBS- and E. coli-inoculated birds, but phagocytic capacity and recruitment were not severely impaired. In spleen samples of IBV-infected animals reduced expression of IL-1β, IL-6, IL-8, IL-10, IL-18 and IFN-γ mRNA was found 1dpi. Our results suggest that enhanced colibacillosis after IBV infection or vaccination is caused at least by altered innate immunity and less by impairment of phagocytic cell function.


title: Intracellular delivery of serum-derived hepatitis C virus

text: Abstract A robust and reliable cell culture system for serum-derived HCV (HCVser) has not been established yet because of the presence of neutralizing antibody and tropism for infection. To overcome this obstacle, we employed a lipid-mediated protein intracellular delivery reagent (PIDR) that permits internalization of proteins into cells. Although entry of HCVcc was not enhanced by the treatment with PIDR, entry of HCVser into hepatoma cell lines (Huh7 and HepG2) and immortalized primary hepatocytes (Hc and HuS/E2) was significantly enhanced by the PIDR treatment. The entry of HCVser into Huh7 cells in the presence of PIDR was resistant to the neutralization by an anti-hCD81 antibody, suggesting that PIDR is capable of internalizing HCVser in a receptor-independent manner. Interestingly, the PIDR-mediated entry of HCVser and HCVcc was enhanced by the addition of sera from chronic hepatitis C patients but not from healthy donors. In addition, neutralization of HCVcc infection by anti-E2 antibody was canceled by the treatment with PIDR. In conclusion, the PIDR is a valuable tool to get over the obstacle of neutralizing antibodies to internalize HCV into cells and might be useful for the establishment of in vitro propagation HCVser.


title: Indoor air pollution and exposure assessment of the gulf cooperation council countries: A critical review

text: Abstract Indoor air pollution is one of the human health threat problems in the Gulf Cooperation Council (GCC) countries. In these countries, due to unfavorable meteorological conditions, such as elevated ambient temperature, high relative humidity, and natural events such as dust storms, people spend a substantial amount of their time in indoor environments. In addition, production of physical and biological aerosols from air conditioners, cooking activities, burning of Arabian incense, and overcrowding due to pilgrimage programs are common causes of low quality indoor air in this region. Thus, due to infiltration of outdoor sources as well as various indoor sources, people living in the GCC countries are highly exposed to indoor air pollutants. Inhalation of indoor air pollutants causes mortalities and morbidities attributed to cardiorespiratory, pulmonary, and lung cancer diseases. Hence, the aim of this review study is to provide a summary of the major findings of indoor air pollution studies in different microenvironments in six GCC countries. These include characterization of detected indoor air pollutants, exposure concentration levels, source identifications, sustainable building designs and ventilation systems, and the mitigation strategies. To do so, >130 relevant indoor air pollution studies across the GCC countries were critically reviewed. Particulate matters (PM10 and PM2.5), total volatile organic compounds (TVOCs), carbon dioxide (CO2), sulfur dioxide (SO2), nitrogen dioxide (NO2), and heavy metals were identified as the reported indoor air pollutants. Apart from them, indoor Radon and bioaerosols were studied only in specific GCC countries. Thus, future studies should also focus on the investigation of emerging indoor air pollutants, such as ultrafine and nanoparticles and their associated health effects. Furthermore, studies on the mitigation of indoor air pollution through the development of advanced air purification and ventilation systems could improve the indoor air quality (IAQ) in the GCC region.


title: Are We Harming Cancer Patients by Delaying Their Cancer Surgery During the COVID-19 Pandemic?

text: Supplemental Digital Content is available in the text


title: Chimeric influenza-virus-like particles containing the porcine reproductive and respiratory syndrome virus GP5 protein and the influenza virus HA and M1 proteins

text: Both porcine reproductive and respiratory syndrome and swine influenza are acute, highly contagious swine diseases. These diseases pose severe threats for the swine industry and cause heavy economic losses worldwide. In this study, we have developed a chimeric virus-like particle (VLP) vaccine candidate for porcine reproductive and respiratory syndrome virus (PRRSV) and H3N2 influenza virus and investigated its immunogenicity in mice. The HA and M1 proteins from the H3N2 influenza virus and the PRRSV GP5 protein fused to the cytoplasmic and transmembrane domains of the NA protein were both incorporated into the chimeric VLPs. Analysis of the immune responses showed that the chimeric VLPs elicited serum antibodies specific for both PRRSV GP5 and the H3N2 HA protein, and they stimulated cellular immune responses compared to the responses to equivalent amounts of inactivated viruses. Taken together, the results suggested that the chimeric VLP vaccine represents a potential strategy for the development of a safe and effective vaccine to control PRRSV and H3N2 influenza virus.


title: Lactobacillus GG Does Not Affect D‐Lactic Acidosis in Diarrheic Calves, in a Clinical Setting

text: D‐lactate, produced by gastrointestinal fermentation, is a major contributor to metabolic acidosis in diarrheic calves. Lactobacillus rhamnosus GG survives gastrointestinal transit in the neonatal calf and does not produce D‐lactate. To determine whether this probiotic reduces gastrointestinal D‐lactate production or severity of diarrhea or both, 48 calves (mean, 11 days old; range, 2–30 days) admitted to the clinic for treatment of diarrhea were randomly allocated to 2 groups. The experimental group was given Lactobacillus rhamnosus GG (1×10(11) cfu/d) PO, dissolved in milk or oral electrolyte solution, in addition to clinic treatment protocols; the other group served as a control. Serum and fecal samples were obtained at admission and at 24 and 48 hours after initial administration of Lactobacillus rhamnosus GG. All samples were analyzed for D‐and L‐lactate by using high‐pressure liquid chromatography. Feces were also analyzed for pathogens, Lactobacillus rhamnosus GG recovery, and dry matter. D‐lactic acidemia (>3 mmol/L) was present in 37/48 calves at admission. Lactobacillus rhamnosus GG was recovered in the feces of 13 experimental calves and 0 control calves 24 hours after administration. No difference in serum or fecal D‐ or L‐lactate between the groups was detected at any time point. After therapy, D‐lactic acidosis was absent at 48 hours in all but 1 calf. No relation between fecal pathogen (viral, bacterial, or protozoal) and degree of D‐lactic acidosis was observed. The reduction in mortality and greater fecal dry matter in Lactobacillus rhamnosus GG‐treated calves was not statistically significant.


Neuraxial Procedures in COVID-19–Positive Parturients: A Review of Current Reports


Palivizumab use during respiratory syncytial virus outbreak in the neonatal intensive care unit


title: Characterization of the complete genome, antigenicity, pathogenicity, tissue tropism, and shedding of a recombinant avian infectious bronchitis virus with a ck/CH/LJL/140901-like backbone and an S2 fragment from a 4/91-like virus

text: Abstract In this study, we isolated an infectious bronchitis virus, designated I1101/16, from broiler breeders in China. Analysis of the S1 gene showed that isolate I1101/16 was genetically close to strain ck/CH/LJL/140901, which belongs to the TW I genotype (also known as lineage GI-7 based on the recent IBV classification), however the S2 gene showed genetic diversity comparing to that of S1 gene. Comparison of the genomic sequences showed that the genome of isolate I1101/16 was similar to that of strain ck/CH/LJL/140901 from the 5′ end of the genome to the 5′ end of the S2 gene and from the 5′ end of the 3a gene to the end of the genome, whereas the remaining parts of the genome sequences were more closely related to those of strain 4/91 than those of ck/CH/LJL/140901, thereby suggesting that recombination might have occurred during the origin of the virus. SimPlot and Bootscan analysis of the complete genomic sequence confirmed this hypothesis, where it showed that isolate I1101/16 arose through recombination events between ck/CH/LJL/140901- and 4/91-like viruses. Isolate I1101/16 and strain ck/CH/LJL/140901 shared identical amino acids in almost all five of their B cell epitopes, but the two viruses had a serotype relatedness value of 65, which is well below 80, i.e., the lower cutoff value for viruses of the same serotype. In addition, pathogenicity tests demonstrated that isolate I1101/16 was more pathogenic to 1-day-old specific-pathogen-free chickens than strain ck/CH/LJL/140901, according to analysis of the clinical signs, whereas strain ck/CH/LJL/140901 exhibited prolonged replication and shedding after challenge compared with isolate I1101/16. This study did not provide evidence that recombination can directly alter the antigenicity, virulence, replication, shedding, and tissue tropism of a virus, but it did show that recombination events are likely to be major determinants of viral evolution.