title: Ebola in the Eastern Democratic Republic of Congo: One Health approach to infectious disease control

text: The Democratic Republic of Congo (DRC) is facing its tenth outbreak of Ebola virus disease (EVD), in North-Kivu and Ituri provinces. This is the second most deadly EVD outbreak in history, after the one that occurred in West Africa in 2014. The DRC Ministry of Health (MoH), supported by the World Health Organization (WHO) and a range of regional and international partners, are implementing EVD response plans in these affected areas such as screening of suspect cases at points of entry, case detection, contact tracing, laboratory testing, case management and infection prevention and control, safe and dignified burials, ring vaccination (this involves vaccination of infected individuals, direct contacts of infected individuals and contacts of their contacts), and therapeutics, community mobilization and free access to healthcare services. Despite these efforts, there has been a sharp rise in the number of confirmed cases within the identified affected areas, and due to a number of challenges unique to DRC, there has been an expansion in the geographical extent of transmission. The significance of the proximity of these regions to wildlife and the Virunga National Park is questionable in the EVD transmission dynamics. The close interaction between human, animal, and environmental factors, in combination with high population movement due to regular rebel attacks in these regions, suggest the need for the integration of the One Health approach in the holistic response plans for control and prevention of EVD. This paper seeks to highlight the implications and importance of a One Health–based approach into the infectious diseases control program implementation in DRC.


Severe acute respiratory syndrome (sars) in children


Coronavirus Disease 2019 (COVID-19) and Radiology Education—Strategies for Survival


title: Understanding the symptoms of the common cold and influenza

text: Summary The common cold and influenza (flu) are the most common syndromes of infection in human beings. These diseases are diagnosed on symptomatology, and treatments are mainly symptomatic, yet our understanding of the mechanisms that generate the familiar symptoms is poor compared with the amount of knowledge available on the molecular biology of the viruses involved. New knowledge of the effects of cytokines in human beings now helps to explain some of the symptoms of colds and flu that were previously in the realm of folklore rather than medicine—eg, fever, anorexia, malaise, chilliness, headache, and muscle aches and pains. The mechanisms of symptoms of sore throat, rhinorrhoea, sneezing, nasal congestion, cough, watery eyes, and sinus pain are discussed, since these mechanisms are not dealt with in any detail in standard medical textbooks.


title: Host factors and viral factors associated with severity of human rhinovirus infant respiratory illness

text: BACKGROUND: Risk factors for severe human rhinovirus (HRV) associated infant illness are unknown. OBJECTIVES: To examine the role of HRV in infant respiratory illness, and assess viral and host risk factors for HRV disease severity. METHODS: We utilized a prospective cohort of term, previously healthy infants enrolled during an inpatient or outpatient visit for acute upper or lower respiratory illness during fall-spring months 2004-2008. Illness severity was determined using an ordinal bronchiolitis severity score with higher scores indicating more severe disease. HRV was identified by real-time RT-PCR. The VP4/VP2 region from HRV positive specimens was sequenced to determine species. RESULTS: Of 630 infants with bronchiolitis or URI, 162 (26%) had HRV; HRV was associated with 18% of bronchiolitis and 47% of URI. Among infants with HRV, 104 (64%) had HRV alone. Host factors associated with more severe HRV illness included maternal and family history of atopy (median score 3.5, IQR [1.0-7.8] vs. 2.0 [1.0-5.2], and 3.5 [1.0-7.5] vs.2.0 [0-4.0]). In adjusted analyses maternal history of atopy conferred an increase in risk for more severe HRV bronchiolitis (OR=2.39, 95% CI:1.14-4.99, p=0.02). In a similar model, maternal asthma was also associated with greater HRV bronchiolitis severity (OR=2.49, 95% CI: 1.10-5.67, p=0.03). Among HRV, 35% were HRVA, 6% HRVB, and 30% HRVC. CONCLUSION: HRV was a frequent cause of bronchiolitis and URI among previously healthy term infants requiring hospitalization or unscheduled outpatient visits. Substantial genetic diversity was seen amongst the HRV, and predominant groups varied by season and year. Host factors including maternal atopy were associated with more severe infant HRV illness.


title: Surface vimentin is critical for the cell entry of SARS-CoV

text: BACKGROUND: Severe acute respiratory syndrome coronavirus (SARS-CoV) caused a global panic due to its high morbidity and mortality during 2002 and 2003. Soon after the deadly disease outbreak, the angiotensin-converting enzyme 2 (ACE2) was identified as a functional cellular receptor in vitro and in vivo for SARS-CoV spike protein. However, ACE2 solely is not sufficient to allow host cells to become susceptible to SARS-CoV infection, and other host factors may be involved in SARS-CoV spike protein-ACE2 complex. RESULTS: A host intracellular filamentous cytoskeletal protein vimentin was identified by immunoprecipitation and LC-MS/MS analysis following chemical cross-linking on Vero E6 cells that were pre-incubated with the SARS-CoV spike protein. Moreover, flow cytometry data demonstrated an increase of the cell surface vimentin level by 16.5 % after SARS-CoV permissive Vero E6 cells were treated with SARS-CoV virus-like particles (VLPs). A direct interaction between SARS-CoV spike protein and host surface vimentin was further confirmed by far-Western blotting. In addition, antibody neutralization assay and shRNA knockdown experiments indicated a vital role of vimentin in cell binding and uptake of SARS-CoV VLPs and the viral spike protein. CONCLUSIONS: A direct interaction between vimentin and SARS-CoV spike protein during viral entry was observed. Vimentin is a putative anti-viral drug target for preventing/reducing the susceptibility to SARS-CoV infection.


title: Defective RNAs of clover yellow mosaic virus encode nonstructural/coat protein fusion products

text: Abstract A small group of 1.2-kb RNAs present on polyribosoes from clover yellow mosaic virus (CYMV)-infected tissue contains sequences from the genomic RNA (gRNA) of CYMV and is encapsidated by CYMV coat protein. Some features of these RNAs suggest that they are similar to defective interfering (DI) RNAs, and would be the first to be reported for the potexvirus group. The prototype 1.2-kb RNA is 1172 nucleotides in length excluding a probable poly(A) tail and is composed of two noncontiguous regions corresponding to 757 nucleotides of the 5′ and 415 nucleotides of the 3′ terminal of CYMV's gRNA. The sequence of the prototype 1.2-kb RNA reveals that the two terminal gRNA regions present in this RNA encode a single open reading frame (ORF) joining the N-terminus of the 191-kDa nonstructural product and the C-terminus of the coat protein to form a 35-kDa 191-kDa/coat protein fusion product. The coding properties of this prototype RNA have been confirmed by translation in vitro of native and synthetic transcripts of the 1.2-kb RNAs, both of which direct the synthesis of the anticipated 35-kDa product which reacts with anti-CYMV antiserum. Three additional 1.2-kb RNA species, each of which contains a unique junction site, have been characterized. In all cases, a fusion ORF encoding a 191-kDa/coat protein fusion product is encoded on the RNA. The presence of a fusion ORF in all members of the 1.2-kb RNA species analyzed suggests that maintenance of this ORF may be important for the survival of this class of RNA within the plant. This coding strategy represents a novel property of plant virus defective RNAs.


title: Characterization of infectious aerosols in health care facilities: An aid to effective engineering controls and preventive strategies()()()

text: Assessment of strategies for engineering controls for the prevention of airborne infectious disease transmission to patients and to health care and related workers requires consideration of the factors relevant to aerosol characterization. These factors include aerosol generation, particle sizes and concentrations, organism viability, infectivity and virulence, airflow and climate, and environmental sampling and analysis. The major focus on attention to engineering controls comes from recent increases in tuberculosis, particularly the multidrug-resistant varieties in the general hospital population, the severely immunocompromised, and those in at-risk and confined environments such as prisons, long-term care facilities, and shelters for the homeless. Many workers are in close contact with persons who have active, undiagnosed, or insufficiently treated tuberculosis. Additionally, patients and health care workers may be exposed to a variety of pathogenic human viruses, opportunistic fungi, and bacteria. This report therefore focuses on the nature of infectious aerosol transmission in an attempt to determine which factors can be systematically addressed to result in proven, applied engineering approaches to the control of infectious aerosols in hospital and health care facility environments. The infectious aerosols of consideration are those that are generated as particles of respirable size by both human and environmental sources and that have the capability of remaining viable and airborne for extended periods in the indoor environment. This definition precludes skin and mucous membrane exposures occurring from splashes (rather than true aerosols) of blood or body fluids containing infectious disease agents. There are no epidemiologic or laboratory studies documenting the transmission of bloodborne virus by way of aerosols. (AJIC Am J Infect Control 1998;26:453-64)


title: Programmed ribosomal frameshifting in decoding the SARS-CoV genome

text: Abstract Programmed ribosomal frameshifting is an essential mechanism used for the expression of orf1b in coronaviruses. Comparative analysis of the frameshift region reveals a universal shift site U_UUA_AAC, followed by a predicted downstream RNA structure in the form of either a pseudoknot or kissing stem loops. Frameshifting in SARS-CoV has been characterized in cultured mammalian cells using a dual luciferase reporter system and mass spectrometry. Mutagenic analysis of the SARS-CoV shift site and mass spectrometry of an affinity tagged frameshift product confirmed tandem tRNA slippage on the sequence U_UUA_AAC. Analysis of the downstream pseudoknot stimulator of frameshifting in SARS-CoV shows that a proposed RNA secondary structure in loop II and two unpaired nucleotides at the stem I–stem II junction in SARS-CoV are important for frameshift stimulation. These results demonstrate key sequences required for efficient frameshifting, and the utility of mass spectrometry to study ribosomal frameshifting.


title: Generating translatable evidence to improve patient care; the contribution of human factors.

text: Abstract Generating translatable evidence to improve patient care has proved challenging in reproductive medicine, with many “add-on” treatments in routine assisted conception clinical practice, which have not been reliably tested. This has consequences for patient care; specifically, IVF pregnancy rates have not improved. A change of culture is required in our profession, from indiscriminately applying the latest “add-on”, to whole-scale participation in generating reliable translatable evidence.


title: A loophole in international quarantine procedures disclosed during the SARS crisis

text: Summary This study describes a loophole in the international quarantine system during the recent Asian severe acute respiratory syndrome (SARS) outbreak. Specifically, that of travelers disguising symptoms of respiratory tract infection at international airports, in order to board aircraft to return to their home countries—notwithstanding the infection risks this involves to others. High medical fees for treatment to non-residents in epidemic areas were found to be the main cause for this behaviour. This phenomenon revealed a loophole in the control mechanisms of international quarantine procedures, letting travelers carrying a highly contagious virus slip by undetected and causing possible multi-country outbreaks of communicable diseases. Clinical evidence collected from medical records at medical centers can highlight this oversight.


title: American Indian Reservations and COVID-19: Correlates of Early Infection Rates in the Pandemic

text: To determine the household and community characteristics most closely associated with variation in COVID-19 incidence on American Indian reservations in the lower 48 states. DESIGN: Multivariate analysis with population weights. SETTING: Two hundred eighty-seven American Indian Reservations and tribal homelands (in Oklahoma) and, as of April 10, 2020, 861 COVID-19 cases on these reservation lands. MAIN OUTCOME MEASURES: The relationship between rate per 1000 individuals of publicly reported COVID-19 cases at the tribal reservation and/or community level and average household characteristics from the 2018 5-Year American Community Survey records. RESULTS: By April 10, 2020, in regression analysis, COVID-19 cases were more likely by the proportion of homes lacking indoor plumbing (10.83, P = .001) and were less likely according to the percentage of reservation households that were English-only (−2.43, P = .03). Household overcrowding measures were not statistically significant in this analysis (−6.40, P = .326). CONCLUSIONS: Failure to account for the lack of complete indoor plumbing and access to potable water in a pandemic may be an important determinant of the increased incidence of COVID-19 cases. Access to relevant information that is communicated in the language spoken by many reservation residents may play a key role in the spread of COVID-19 in some tribal communities. Household overcrowding does not appear to be associated with COVID-19 infections in our data at the current time. Previous studies have identified household plumbing and overcrowding, and language, as potential pandemic and disease infection risk factors. These risk factors persist. Funding investments in tribal public health and household infrastructure, as delineated in treaties and other agreements, are necessary to protect American Indian communities.


title: Voices from a Pandemic

text: Our many voices form the human chorus. Here, we present six diverse perspectives that share a common thread: how COVID-19 has changed our lives. We hear about the difficulties of providing palliative care on the front, the challenges patients seeking gender-affirming surgeries face, the loss of rituals built into healthcare visits, the pivots researchers take to study SARS-CoV-2, as well as the unique mental health impact of an ongoing trauma. These are but a few of the myriad voices that represent our COVID-19 collective. Yet they highlight a reality: a pandemic not only touches all people, but also elicits responses we never quite imagined.


title: Antiviral strategies to control calicivirus infections

text: Abstract Caliciviridae are human or non-human pathogenic viruses with a high diversity. Some members of the Caliciviridae, i.e. human pathogenic norovirus or rabbit hemorrhagic disease virus (RHDV), are worldwide emerging pathogens. The norovirus is the major cause of viral gastroenteritis worldwide, accounting for about 85% of the outbreaks in Europe between 1995 and 2000. In the United States, 25 million cases of infection are reported each year. Since its emergence in 1984 as an agent of fatal hemorrhagic diseases in rabbits, RHDV has killed millions of rabbits and has been dispersed to all of the inhabitable continents. In view of their successful and apparently increasing emergence, the development of antiviral strategies to control infections due to these viral pathogens has now become an important issue in medicine and veterinary medicine. Antiviral strategies have to be based on an understanding of the epidemiology, transmission, clinical symptoms, viral replication and immunity to infection resulting from infection by these viruses. Here, we provide an overview of the mechanisms underlying calicivirus infection, focusing on the molecular aspects of replication in the host cell. Recent experimental data generated through an international collaboration on structural biology, virology and drug design within the European consortium VIZIER is also presented. Based on this analysis, we propose antiviral strategies that may significantly impact on the epidemiological characteristics of these highly successful viral pathogens.


title: The next generation of antimicrobial peptides (AMPs) as molecular therapeutic tools for the treatment of diseases with social and economic impacts

text: Anti-infective drugs have had a key role in the contemporary world, contributing to dramatically decrease mortality rates caused by infectious diseases worldwide. Antimicrobial peptides (AMPs) are multifunctional effectors of the innate immune system of mucosal surfaces and present antimicrobial activity against a range of pathogenic viruses, bacteria, and fungi. However, the discovery and development of new antibacterial drugs is a crucial step to overcome the great challenge posed by the emergence of antibiotic resistance. In this review, we outline recent advances in the development of novel AMPs with improved antimicrobial activities that were achieved through characteristic structural design. In addition, we describe recent progress made to overcome some of the major limitations that have hindered peptide biosynthesis.


title: Nucleolar proteomics and viral infection

text: Recent advances in proteomics have been combined with traditional methods for isolation of nucleoli from mammalian and plant cells. This approach has confirmed the growing body of data showing a wide role for the nucleolus in eukaryotic cell biology beyond ribosome generation into many areas of cell function from regulation of the cell cycle, modulation of the cell stress response to innate immune responses. This has been reflected in the growing body of evidence that viruses specifically target the nucleolus by sequestering cellular nucleolar proteins or by targeting viral proteins to the nucleolus in order to maximise viral replication. This review covers those key areas and looks at the latest approaches using high‐throughput quantitative proteomics of the nucleolus in virus infected cells to gain an insight into the role of this fascinating compartment in viral infection.


title: COVID-19 pneumonia manifestations at the admission on chest ultrasound, radiographs, and CT: single-center study and comprehensive radiologic literature review

text: PURPOSE: To investigate the imaging features of emerging COVID-19 pneumonia on chest ultrasound (US), radiographs (CXR) and computed tomography (CT) examinations performed at admission and to provide a comprehensive radiological literature review on ongoing radiological data from recent publications. MATERIALS AND METHODS: In this retrospective single-center study, we enrolled consecutive patients from February 15, 2020, to March 15, 2020, with laboratory-confirmed SARS-CoV-2 hospitalized in Valduce Hospital (Como, Italy). Multi-modality imaging findings were evaluated and compared. Literature research was conducted through a methodical search on Pubmed and Embase databases. RESULTS: Fifty-eight patients (36 men, 22 women; age range, 18–98 years) were included in the study. Among these, chest US, CXR, and CT were performed respectively in twenty-two, thirty-two and forty-two patients. Lung US findings were consistent with diffuse B lines (100%) and subpleural consolidations (27.3%). CXR showed prevalent manifestations of consolidations (46.9%) and hazy increased opacities (37.5%). Typical CT features included bilateral and multilobar ground-glass opacities (GGO) with (59.5%) and without (35.7%) consolidations having a predominantly peripheral distribution (64.3%). Other imaging features included crazy paving pattern (57.1%), fibrous stripes (50%), subpleural lines (35.7%), architectural distortion (28.6%), air bronchogram sign (26.2%), vascular thickening (23.8%) and nodules (2.4%). Also, enlarged lymph nodes (14.3 %) and pleural effusion (7.1%) were observed. The literature review identified twenty-six original studies supporting our imaging chest findings. CONCLUSION: The spectrum of chest imaging manifestations of COVID-19 pneumonia upon admission includes B-lines and consolidations on US, consolidations and hazy increased opacities on CXR, and multifocal GGO with consolidations on CT.


title: Impact of rhinovirus nasopharyngeal viral load and viremia on severity of respiratory infections in children

text: There are few and partially discordant data regarding nasopharyngeal rhinovirus (RV) load and viremia, and none of the published studies evaluated the two variables together. The aim of this study was to provide new information concerning the clinical relevance of determining nasopharyngeal viral load and viremia when characterising RV infection. Nasopharyngeal swabs were obtained from 251 children upon their admission to hospital because of fever and signs and symptoms of acute respiratory infection in order to identify the virus and determine its nasopharyngeal load, and a venous blood sample was taken in order to evaluate viremia. Fifty children (19.9 %) had RV-positive nasopharyngeal swabs, six (12 %) of whom also had RV viremia: RV-C in four cases (66.6 %), and RV-A and RV-B in one case each. The RV nasopharyngeal load was significantly higher in the children with RV viremia (p < 0.001), who also had a higher respiratory rate (p = 0.02), white blood cell counts (p = 0.008) and C-reactive protein levels (p = 0.006), lower blood O(2) saturation levels (P = 0.005), and more often required O(2) therapy (p = 0.009). The presence of RV viremia is associated with a significantly higher nasopharyngeal viral load and more severe disease, which suggests that a high nasopharyngeal viral load is a prerequisite for viremia, and that viremia is associated with considerable clinical involvement.


title: Corrigendum to “Middle East Respiratory Syndrome Coronavirus Outbreak in the Republic of Korea, 2015” [Volume 6, Issue 4, August 2015, 269–278]

text: [This corrects the article DOI: 10.1016/j.phrp.2015.08.006.].


title: Virus-mediated modulation of the host endocrine signaling systems: clinical implications

text: Viruses, which are among the simplest infective pathogens, can produce characteristic endocrine manifestations in infected patients. In addition to the classic modification of the host endocrine system by either direct or indirect destruction of the endocrine organs and/or effects exerted by systemic production of inflammatory and/or stress mediators, recent progress in molecular virology and endocrinology has revealed that virus-encoded molecules might alter the host endocrine-signaling systems by affecting extracellular and/or intracellular signal transduction and hormone sensitivity of host target tissues. Here, we provide a brief overview of such viral-mediated modulation of host endocrine signaling systems. We propose that virus-encoded molecules and the signaling systems they influence are potential therapeutic targets for the treatment of disorders that are associated with some viral infections.


title: Maintaining the silence: reflections on long-term RNAi

text: Since the demonstration of RNA interference (RNAi) in mammalian cells, considerable research and financial effort has gone towards implementing RNAi as a viable therapeutic platform. RNAi is, without doubt, the most promising strategy for the treatment of human genetic disorders. Because many of the targets proposed for RNAi therapy require chronic treatment, researchers agree that the emphasis must now be placed on the safe and long-term application of RNAi drugs to reap the benefits at last.


title: Effectiveness of composting as a biosecure disposal method for porcine epidemic diarrhea virus (PEDV)-infected pig carcasses

text: BACKGROUND: Porcine epidemic diarrhea virus (PEDV) is an enteric disease of swine that has emerged as a worldwide threat to swine herd health and production. Substantial research has been conducted to assess viability of the virus on surfaces of vehicles and equipment, in feed and water, and on production building surfaces, but little is known about the persistence in PEDV-infected carcasses and effective disposal methods thereof. This study was conducted to quantify the persistence of PEDV RNA via quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) at various time-temperature combinations and in infected piglet carcasses subjected to composting. Although this method does not distinguish between infectious and noninfectious virus, it is a rapid and sensitive test to evaluate materials for evidence of virus genome. RESULTS: In the first study, PEDV was suspended in cell culture media at 1 × 10(5) TCID50 per sample (1 mL sample size) and subjected to various time and temperature combinations in triplicate including temperatures of 37, 45, 50, 55, 60, 65, 70 °C and exposure times of 0, 1, 2, 3, 4, 5, 7, and 14 days. At all temperatures, viral RNA copies declined over time, with the decline most marked and rapid at 65 and 70 °C. Detectable RNA did persist throughout the trial in all but the most extreme condition, where two of three samples incubated at 70 °C yielded undetectable viral RNA after 14 days. In the second study, PEDV-infected piglet carcasses were subjected to two cycles of composting lasting 36 and 37 days, respectively, for a total compost time of 73 days. Composting was performed in triplicate windrow sections housed inside biosecure, climate-controlled rooms using insulated bins designed to represent a continuous windrow compost pile. Temperatures reached 35–57 °C for 26 days of cycle 1 and 35–45 °C for 3 days of cycle 2. Samples consisting of carbon material with or without decomposed tissue as available per sample site collected at ten locations throughout the cross-section of each windrow section following the primary and secondary compost cycles yielded no detectable viral RNA. CONCLUSIONS: Composting appears to be an effective disposal method for PEDV-infected piglet carcasses under the conditions examined. The combination of time and high temperature of the compost cycle effectively degraded viral RNA in cell culture media that should provide optimum stability. Complex compost material matrices collected from windrow sections yielded undetectable PEDV RNA by qRT-PCR after one 36-day compost cycle despite incomplete decomposition of soft tissue.


title: Ribavirin is not effective against Crimean–Congo hemorrhagic fever: observations from the Turkish experience

text: Summary Crimean–Congo hemorrhagic fever (CCHF) is a viral infection associated with a high mortality rate. Ribavirin is the only drug used in the treatment of this disease. Studies investigating the effectiveness of ribavirin in CCHF have been retrospective and to date have included only a small number of cases. In recent years, due to climate changes, the number of cases of CCHF in Turkey has increased, and experience in the treatment of CCHF has improved. Several studies have evaluated the efficacy of ribavirin in Turkey, including one randomized controlled trial and two studies with a large number of cases. In these studies, ribavirin therapy was not shown to decrease mortality rates; the mortality rate was 2–9% in patients treated with ribavirin and 5.6–11% in those who were not treated with this drug. These findings suggest that patients with CCHF should be followed with supportive care only until randomized controlled trials with larger groups have been conducted.


title: A Practical Guide to the Diagnosis, Treatment, and Prevention of Neonatal Infections

text: Neonatal infections continue to cause morbidity and mortality in infants. Among approximately 400,000 infants followed nationally, the incidence rates of early-onset sepsis infection within 3 days of life are 0.98 cases per 1000 live births. Newborn infants are at increased risk for infections because they have relative immunodeficiency. This article provides evidence-based practical approaches to the diagnosis, management, and prevention of neonatal infections.


COVID testing before every endoscopy: Is India ready for primetime?


title: La salud en todas las políticas. Tiempo de crisis, ¿tiempo de oportunidades? Informe SESPAS 2010

text: A cost-effective and equitable health policy design involves tackling social determinants of health and becoming aware of the impact of non-healthcare policies. Structural changes in institutional organization, targeted to create a new space for network governance, are required. Some factors tend to delay this process, such as institutional inertia, poor coordination and cooperation mechanisms among sectors, levels of government and agents, the consolidation of the biomedical paradigm reinforced by lobbies and cultural patterns, and inadequate collaboration among experts, practitioners and health authorities. However, these factors may be counterbalanced by political will and by the knowledge provided by domestic and foreign experiences. The present situation arising from the recent public health crises, and particularly from the current economic depression, offers a major chance to introduce the needed changes in health oriented actions.


COVID-19 pandemic alert: time to focus on lung health-Beijing call to action for lung health promotion


title: Room-temperature-storable PCR Mixes for SARS-CoV-2 Detection

text: OBJECTIVES: A novel coronavirus (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) emerged in late 2019, causing an outbreak of pneumonia [coronavirus disease 2019 (COVID-19)] globally. Although the use of ready-made reaction mixes can enable more rapid PCR-based diagnosis of COVID-19, the need to transport and store these mixes at low temperatures presents challenges to already overburdened logistics networks. METHODS: Here, we present an optimized freeze-drying procedure that allows SARS-CoV-2 PCR mixes to be transported and stored at ambient temperatures, without loss of activity. Additive-supplemented PCR mixes were freeze-dried. The residual moisture of the freeze-dried PCR mixes was measured by Karl-Fischer titration. RESULTS: We found that freeze-dried PCR mixes with ∼1.2% residual moisture are optimal for storage, transport, and reconstitution. The sensitivity, specificity, and repeatability of the freeze-dried reagents were similar to those of freshly prepared, wet reagents. The freeze-dried mixes retained activity at room temperature (18∼25℃) for 28 days, and for 14 and 10 days when stored at 37℃ and 56℃, respectively. CONCLUSION: The uptake of this approach will ease logistical challenges faced by transport networks and make more cold storage space available at diagnosis and hospital laboratories.


title: Dioctophyma renale in a dog: Clinical diagnosis and surgical treatment

text: Abstract This study reports a case of parasitism by the giant kidney worm, Dioctophyma renale, diagnosed in the right kidney of a domestic dog. An adult female German Shepherd was attended with clinical history of prostration and hyporexia. The hemogram showed changes compatible with an inflammatory process, for that reason, an abdominal ultrasound was requested. Ultrasound image suggested the presence of D. renale in the right kidney. The diagnosis was confirmed after urinalysis due to the presence of dioctophymas ova in the urinary sediment. Surgical treatment was made and the animal had an excellent recovery after the nephrectomy was performed. Generally, in almost all cases, parasitism by D. renale in domestic dogs is a necropsy finding, nevertheless imaging techniques as sonography and laboratorial exams as urinalysis have been proven to be important tools to achieve diagnosis. The purpose of this study is to report a case of parasitism by D. renale where diagnosis and treatment were made in time to allow the patient's recovery.


title: Generation of potent neutralizing human monoclonal antibodies against cytomegalovirus infection from immune B cells

text: BACKGROUND: Human monoclonal antibodies (mAbs) generated as a result of the immune response are likely to be the most effective therapeutic antibodies, particularly in the case of infectious diseases against which the immune response is protective. Human cytomegalovirus (HCMV) is an ubiquitous opportunistic virus that is the most serious pathogenic agent in transplant patients. The available therapeutic armamentarium (e.g. HCMV hyperimmune globulins or antivirals) is associated with severe side effects and the emergence of drug-resistant strains; therefore, neutralizing human mAb may be a decisive alternative in the prevention of primary and re-activated HCMV infections in these patients. RESULTS: The purpose of this study was to generate neutralizing mAb against HCMV from the immunological repertoire of immune donors. To this aim, we designed an efficient technology relying on two discrete and sequential steps: first, human B-lymphocytes are stimulated with TLR9-agonists and IL-2; second, after both additives are removed, the cells are infected with EBV. Using this strategy we obtained 29 clones secreting IgG neutralizing the HCMV infectivity; four among these were further characterized. All of the mAbs neutralize the infection in different combinations of HCMV strains and target cells, with a potency ~20 fold higher than that of the HCMV hyperimmune globulins, currently used in transplant recipients. Recombinant human monoclonal IgG1 suitable as a prophylactic or therapeutic tool in clinical applications has been generated. CONCLUSION: The technology described has proven to be more reproducible, efficient and rapid than previously reported techniques, and can be adopted at low overall costs by any cell biology laboratory for the development of fully human mAbs for immunotherapeutic uses.


Will we be judged ready for the influenza pandemic?


title: I Can’t Breathe: The Continued Disproportionate Underrepresentation and Effective Exclusion of Black Physicians in the United States Radiation Oncology Workforce

text: Abstract Purpose Black physicians remain disproportionately underrepresented in certain medical specialties, yet comprehensive assessments in Radiation Oncology (RO) are lacking. Our purpose was to report current and historical representation trends for Black physicians in the US RO workforce. Methods Public registries were used to assess significant differences in 2016 representation for US vs RO Black academic full-time faculty, residents, and applicants. Historical changes from 1970-2016 were reported descriptively. Linear regression was used to assess significant changes for Black residents and faculty from 1995-2016. Results In 2016, Blacks represented 3.2% vs 1.5% (P<0.001), 5.6% vs 3.2% (P=0.005), and 6.5% vs 5.4% (P=0.352), of US vs RO faculty, residents, and applicants, respectively. While RO residents nearly doubled from 374 (1974) to 720 (2016), Black residents peaked at 31 in 1984 (5.9%; 31/522) and fell to 23 (3.2%; 23/720) in 2016 across 91 accredited programs; Black US GME trainees nearly doubled over the same time period: 3,506 (1984) to 6,905 (2016). Between 1995-2016, Black US resident representation significantly increased 0.03%/yr, but decreased significantly in RO -0.20%/yr before 2006 and did not change significantly thereafter. Over the same time period, Black US faculty representation significantly increased 0.02%/yr, while Black RO faculty significantly increased 0.07%/yr before 2006, then decreased significantly -0.16%/yr thereafter. The number of Black RO faculty peaked at 37 in 2006 (3.1%; 37/1203) and was 27 (1.5%; 27/1769) in 2016, despite the nearly 1.5-fold increase in the number of both RO faculty and Black US faculty overall (4,169 in 2006 and 6,047 in 2016) during that time period. Conclusions Black physicians remain disproportionately underrepresented in RO despite an increasing available pipeline in the US physician workforce. Deliberate efforts to understand barriers to specialty training and inclusion, along with evidence-based targeted interventions to overcome them, are needed to ensure diversification of the RO physician workforce.


title: COPD exacerbations: defining their cause and prevention

text: Summary Exacerbations of chronic obstructive pulmonary disease (COPD) are episodes of worsening of symptoms, leading to substantial morbidity and mortality. COPD exacerbations are associated with increased airway and systemic inflammation and physiological changes, especially the development of hyperinflation. They are triggered mainly by respiratory viruses and bacteria, which infect the lower airway and increase airway inflammation. Some patients are particularly susceptible to exacerbations, and show worse health status and faster disease progression than those who have infrequent exacerbations. Several pharmacological interventions are effective for the reduction of exacerbation frequency and severity in COPD such as inhaled steroids, long-acting bronchodilators, and their combinations. Non-pharmacological therapies such as pulmonary rehabilitation, self-management, and home ventilatory support are becoming increasingly important, but still need to be studied in controlled trials. The future of exacerbation prevention is in assessment of optimum combinations of pharmacological and non-pharmacological therapies that will result in improvement of health status, and reduction of hospital admission and mortality associated with COPD.


title: Evaluation of Fast Technology Analysis (FTA) Cards as an improved method for specimen collection and shipment targeting viruses associated with Bovine Respiratory Disease Complex

text: Abstract In order to improve the analytic quality of respiratory specimens collected from cattle for nucleic acid-based diagnosis, a study was undertaken to verify realtime PCR efficiency of specimens collected and stabilized on FTA Cards™, filter paper which is treated chemically. Nucleic acids collected using FTA Cards without the need for a cold-chain or special liquid media handling provided realtime PCR results consistent (96.8% agreement, kappa 0.923 [95% CI=0.89–0.96]) with the same specimens collected using traditional viral transport media and shipped on ice using the U.S. Department of Transportation mandated liquid handling requirements. Nucleic acid stabilization on FTA Cards was evaluated over a temperature range (−27°C to +46°C) for up to 14 days to mimic environmental conditions for diagnostic sample handling between collection and processing in a routine veterinary laboratory. No significant difference (P ≥0.05) was observed in realtime PCR cycle threshold values over the temperature range and time storage conditions for Bovine Viral Diarrhea virus, Bovine Respiratory Syncytial virus, Bovine Coronavirus, and Bovine Herpesvirus I. The four viruses evaluated in the study are associated with Bovine Respiratory Disease Complex where improvements in ease and reliability of specimen collection and shipping would enhance the diagnostic quality of specimens collected in the field, and ultimately improve diagnostic efficiency.


title: Legionella: a Fascinating Bacterium Uncovered in the Twentieth Century

text: In July 1976, the American Legion held a conference at the Bellevue Stratford Hotel in Philadelphia, PA, to celebrate the nation's bicentennial. This convention resulted in transmission of a gram-negative bacterium to over 200 attendees, who developed a respiratory illness; 34 deaths were attributed to the infections. An investigation of the illness revealed a bacterium that had not been documented before. The disease became known as Legionnaires' disease, and the etiological agent was subsequently named Legionella pneumophila. This is the story of Legionella, with special emphasis on its ecological niche, the diagnosis of human infection, and its isolation from the environment. There are only a handful of diseases that debuted in the 20th or 21st century. They include Legionnaires' disease (the subject of this review), Lyme disease, AIDS, severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and West Nile virus.


title: Impact of acute psychosocial stress on peripheral blood gene expression pathways in healthy men

text: Abstract We investigated peripheral blood mononuclear cell gene expression responses to acute psychosocial stress to identify molecular pathways relevant to the stress response. Blood samples were obtained from 10 healthy male subjects before, during and after (at 0, 30, and 60min) a standardized psychosocial laboratory stressor. Ribonucleic acid (RNA) was extracted and gene expression measured by hybridization to a 20,000-gene microarray. Gene Set Expression Comparisons (GSEC) using defined pathways were used for the analysis. Forty-nine pathways were significantly changed from baseline to immediately after the stressor (p <0.05), implicating cell cycle, cell signaling, adhesion and immune responses. The comparison between stress and recovery (measured 30min later) identified 36 pathways, several involving stress-responsive signaling cascades and cellular defense mechanisms. These results have relevance for understanding molecular mechanisms of the physiological stress response, and might be used to further study adverse health outcomes of psychosocial stress.


title: Anti-Neoplastic Effects of Gallic Acid, a Major Component of Toona sinensis Leaf Extract, on Oral Squamous Carcinoma Cells

text: Extract of Toona sinensis (TS) has been reported to have various effects on cultured cell lines, including anti-proliferative activity in cancer cells. We have studied the effects of TS on various human oral squamous carcinoma cell lines (HOSCC), including UM1, UM2, SCC-4, and SCC-9. These cell lines were treated with TS leaf extract and screened for viability, apoptosis, necrosis, and apoptotic gene expression. Normal human oral keratinocytes (NHOK) served as a control for cytotoxic assays. Viability of TS-treated HOSCC was reduced, whereas that of NHOK was not affected. FACScan analysis revealed that the leaf extract induced apoptosis or a combination of apoptosis and necrosis, depending on cell type. Microarray and semi-quantitative RT-PCR analysis for apoptotic-related gene expression revealed that 3,4,5-trihydroxybenzoic acid (gallic acid, one of the major bioactive compounds purified from TS extract) up-regulated pro-apoptotic genes such TNF-α, TP53BP2, and GADD45A, and down-regulated the anti-apoptotic genes Survivin and cIAP1, resulting in cell death. This study suggests that gallic acid, the major bioactive compound present, is responsible for the anti-neoplastic effect of Toona sinensis leaf extract.


title: Modulation of iridovirus-induced apoptosis by endocytosis, early expression, JNK, and apical caspase

text: Abstract Chilo iridescent virus (CIV) is the type species for the family Iridoviridae, which are large, isometric, cytoplasmic dsDNA viruses. We examined the mechanism of apoptosis induction by CIV. High CIV doses (CIVXS; 400 μg/ml), UV-irradiated virus (CIVUV; 10 μg/ml) and CVPE (CIV protein extract; 10 μg/ml) induced apoptosis in 60% of treated Choristoneura fumiferana (IPRI-CF-124T) cells. Normal doses of infectious CIV (10 μg/ml) induced apoptosis in only 10% of C. fumiferana (CF) cells. Apoptosis was inhibited by Z-IETD-FMK, an apical caspase inhibitor, indicating that CIV-induced apoptosis requires caspase activity. The putative caspase in CF cells was designated Cf-caspase-i. CIVUV or CVPE enhanced Cf-caspase-i activity by 80% at 24 h relative to mock-treated cells. Since the MAP kinase pathway induces or inhibits apoptosis depending on the context, we used JNK inhibitor SP600125 and demonstrated drastic suppression of CVPE-induced apoptosis. Thus, the JNK signaling pathway is significant for apoptosis in this system. Virus interaction with the cell surface was not sufficient for apoptosis since CIVUV particles bound to polysterene beads failed to induce apoptosis. Endocytosis inhibitors (bafilomycin or ammonium chloride) negated apoptosis induction by CIVUV, CIVXS or CVPE indicating that entry through this mode is required. Given the weak apoptotic response to infectious CIV, we postulated that viral gene expression inhibited apoptosis. CIV infection of cells pretreated with cycloheximide induced apoptosis in 69% of the cells compared to 10% in normal infections. Furthermore, blocking viral DNA replication with aphidicolin or phosphonoacetic acid suppressed apoptosis and Cf-caspase-i activity, indicating that early viral expression is necessary for inhibition of apoptosis, and de novo synthesis of viral proteins is not required for induction. We show for the first time that, in a member of the family Iridoviridae, apoptosis: (i) requires entry and endocytosis of virions or virion proteins, (ii) is inhibited under conditions permitting early viral expression, and (iii) requires the JNK signaling pathway. This is the first report of JNK signal requirement during apoptosis induction by an insect virus.


title: Clinical and epidemiological characteristics of Eimeria infections in first-year grazing cattle

text: Abstract Infections with Eimeria parasites can lead to severe diarrhoea with considerable clinical and economic consequences in first-year grazing stock. To identify and characterise the cause of diarrhoea observed during previous years, 164 animals on 14 dairy farms in northwestern Germany were included in this study. The calves were physically and parasitologically examined prior to turnout and until 21 days post turnout (d.p.t.). Mean animal weights decreased from 194.9kg at the start to 189.3kg bodyweight at the end of the study. In all herds, oocyst counts were very low prior to turnout and increased after the calves had been kept on pasture for at least 7 days. On Day 9 post turnout, 90% and at the end of the study (21d.p.t.) 70% of all animals showed Eimeria-positive faecal samples. During the course of the study, 79 (48.2%) animals passed faecal samples with more than 100,000 oocysts per gram. The predominant species identified was Eimeria alabamensis, which accounted for more than 83% of the oocysts counted. These parasitological findings matched the clinical observations. Diarrhoea was found in 130 (79.3%) of the study animals. At 5d.p.t. and thus prior to the rise of faecal oocyst counts, a significant increase in diarrhoea was recorded. Calves showing diarrhoea excreted statistically significantly more often over 100,000 E. alabamensis oocysts per gram faeces (0.28; p =0.0002) than calves without diarrhoea. Diarrhoea was also found during significantly more study days in animals with high oocyst counts (0.39; p =0.0001). These data indicate that in endemic areas first-year grazing calves must be considered at risk to develop clinical coccidiosis due to E. alabamensis infection during the first 2–3 weeks post turnout.


title: Using Informatics and the Electronic Medical Record to Describe Antimicrobial Use in the Clinical Management of Diarrhea Cases at 12 Companion Animal Practices

text: Antimicrobial drugs may be used to treat diarrheal illness in companion animals. It is important to monitor antimicrobial use to better understand trends and patterns in antimicrobial resistance. There is no monitoring of antimicrobial use in companion animals in Canada. To explore how the use of electronic medical records could contribute to the ongoing, systematic collection of antimicrobial use data in companion animals, anonymized electronic medical records were extracted from 12 participating companion animal practices and warehoused at the University of Calgary. We used the pre-diagnostic, clinical features of diarrhea as the case definition in this study. Using text-mining technologies, cases of diarrhea were described by each of the following variables: diagnostic laboratory tests performed, the etiological diagnosis and antimicrobial therapies. The ability of the text miner to accurately describe the cases for each of the variables was evaluated. It could not reliably classify cases in terms of diagnostic tests or etiological diagnosis; a manual review of a random sample of 500 diarrhea cases determined that 88/500 (17.6%) of the target cases underwent diagnostic testing of which 36/88 (40.9%) had an etiological diagnosis. Text mining, compared to a human reviewer, could accurately identify cases that had been treated with antimicrobials with high sensitivity (92%, 95% confidence interval, 88.1%–95.4%) and specificity (85%, 95% confidence interval, 80.2%–89.1%). Overall, 7400/15,928 (46.5%) of pets presenting with diarrhea were treated with antimicrobials. Some temporal trends and patterns of the antimicrobial use are described. The results from this study suggest that informatics and the electronic medical records could be useful for monitoring trends in antimicrobial use.


title: Selectivity in ISG15 and ubiquitin recognition by the SARS coronavirus papain-like protease

text: The severe acute respiratory syndrome coronavirus papain-like protease (SARS-CoV PLpro) carries out N-terminal processing of the viral replicase polyprotein, and also exhibits Lys48-linked polyubiquitin chain debranching and ISG15 precursor processing activities in vitro. Here, we used SDS–PAGE and fluorescence-based assays to demonstrate that ISG15 derivatives are the preferred substrates for the deubiquitinating activity of the PLpro. With k(cat)/K(M) of 602,000 M(−1) s(−1), PLpro hydrolyzes ISG15-AMC 30- and 60-fold more efficiently than Ub-AMC and Nedd8-AMC, respectively. Data obtained with truncated ISG15 and hybrid Ub/ISG15 substrates indicate that both the N- and C-terminal Ub-like domains of ISG15 contribute to this preference. The enzyme also displays a preference for debranching Lys48- over Lys63-linked polyubiquitin chains. Our results demonstrate that SARS-CoV PLpro can differentiate between ubiquitin-like modifiers sharing a common C-terminal sequence, and that the debranching activity of the PLpro is linkage type selective. The potential structural basis for the demonstrated specificity of SARS-CoV PLpro is discussed.


title: Comparative analysis of synovial fluid and plasma proteomes in juvenile arthritis – Proteomic patterns of joint inflammation in early stage disease

text: Abstract Synovial fluid is a potential source of novel biomarkers for many arthritic disorders involving joint inflammation, including juvenile idiopathic arthritis. We first compared the distinctive protein ‘fingerprints’ of local inflammation in synovial fluid with systemic profiles within matched plasma samples. The synovial fluid proteome at the time of joint inflammation was then evaluated across clinical subgroups to identify early disease associated proteins. We measured the synovial fluid and plasma proteomes using the two-dimensional fluorescence difference gel electrophoresis approach. Image analysis software was used to highlight the expression levels of joint and subgroup associated proteins across the study cohort (n =32). A defined subset of 30 proteins had statistically significant differences (p <0.05) between sample types such that synovial fluid could be differentiated from plasma. Furthermore distinctive synovial proteome expression patterns segregate patient subgroups. Protein expression patterns localized in the chronically inflamed joint therefore have the potential to identify patients more likely to suffer disease which will spread from a single joint to multiple joints. The proteins identified could act as criteria to prevent disease extension by more aggressive therapeutic intervention directed at an earlier stage than is currently possible.


title: Antimicrobial resistance and virulence factors in Escherichia coli from swedish dairy calves

text: BACKGROUND: In Sweden, knowledge about the role of enteropathogenic Escherichia coli in neonatal calf diarrhea and the occurrence of antimicrobial resistance in E. coli from young calves is largely unknown. This has therapeutic concern and such knowledge is also required for prudent use of antimicrobials. METHODS: In a case control study Esherichia coli isolated from faecal samples from dairy calves were phenotyped by biochemical fingerprinting and analyzed for virulence genes by PCR. Antimicrobial susceptibility was tested by determination of minimum inhibitory concentration (MIC). Farm management data were collected and Fisher's exact test and univariable and multivariable logistic regression analysis were performed. RESULTS: Of 95 E. coli tested for antimicrobial susceptibility 61% were resistant to one or more substances and 28% were multi-resistant. The virulence gene F5 (K99) was not found in any isolate. In total, 21 out of 40 of the investigated virulence genes were not detected or rarely detected. The virulence genes espP, irp, and fyuA were more common in resistant E. coli than in fully susceptible isolates (P < 0.05). The virulence gene terZ was associated with calf diarrhea (P ≤ 0.01). The participating 85 herds had a median herd size of 80 lactating cows. Herds with calf diarrhea problems were larger (> 55 cows; P < 0.001), had higher calf mortality (P ≤ 0.01) and calf group feeders were more in use (P < 0.05), compared to herds without calf diarrhea problems. There was no association between calf diarrhea and diversity of enteric E. coli. CONCLUSIONS: Antimicrobial resistance was common in E. coli from pre-weaned dairy calves, occurring particularly in calves from herds experiencing calf diarrhea problems. The results indicate that more factors than use of antimicrobials influence the epidemiology of resistant E. coli. Enteropathogenic E. coli seems to be an uncommon cause of neonatal calf diarrhea in Swedish dairy herds. In practice, calf diarrhea should be regarded holistically in a context of infectious agents, calf immunity, management practices etc. We therefore advice against routine antimicrobial treatment and recommend that bacteriological cultures, followed by testing for antimicrobial susceptibility and for virulence factors, are used to guide decisions on such treatment.


title: One Health research and training and government support for One Health in South Asia

text: INTRODUCTION: Considerable advocacy, funding, training, and technical support have been provided to South Asian countries to strengthen One Health (OH) collaborative approaches for controlling diseases with global human pandemic potential since the early 2000s. It is essential that the OH approach continues to be strengthened given South Asia is a hot spot for emerging and endemic zoonotic diseases. The objectives of this article are to describe OH research and training and capacity building activities and the important developments in government support for OH in these countries to identify current achievements and gaps. MATERIALS AND METHODS: A landscape analysis of OH research, training, and government support in South Asia was generated by searching peer-reviewed and grey literature for OH research publications and reports, a questionnaire survey of people potentially engaged in OH research in South Asia and the authors’ professional networks. RESULTS: Only a small proportion of zoonotic disease research conducted in South Asia can be described as truly OH, with a significant lack of OH policy-relevant research. A small number of multisectoral OH research and OH capacity building programmes were conducted in the region. The governments of Bangladesh and Bhutan have established operational OH strategies, with variable progress institutionalising OH in other countries. Identified gaps were a lack of useful scientific information and of a collaborative culture for formulating and implementing integrated zoonotic disease control policies and the need for ongoing support for transdisciplinary OH research and policy-relevant capacity building programmes. DISCUSSION: Overall we found a very small number of truly OH research and capacity building programmes in South Asia. Even though significant progress has been made in institutionalising OH in some South Asian countries, further behavioural, attitudinal, and institutional changes are required to strengthen OH research and training and implementation of sustainably effective integrated zoonotic disease control policies.


title: Transitioning a Surgery Practice to Telehealth During COVID-19

text: The spread of COVID-19 has challenged practices across the United States to dramatically change inpatient and outpatient practices to focus on containing spread of the virus and accommodate the anticipated increase in volume of acute illness. Despite this transition, practices will need to continue to accommodate evaluation of newly diagnosed malignancies, routine postoperative visits, and acute postoperative issues. Building on an existing telemedicine framework, we aim to describe rapid transition in our outpatient care to a telehealth model in a general thoracic surgery practice during COVID-19.


title: Diarylheptanoids as nutraceutical: A review

text: Abstract Phenolic compounds are naturally occurring compounds present ubiquitously in plants. They have potential health benefits and substantiate evidence for their nutraceutical applications. Diarylheptanoids are part of the broad class of plant phenolics with structurally divergent compounds. They have been used in traditional medicines and homemade remedies to treat various ailments, as organoleptic additives in foods, and also for aesthetic purposes. With their potential therapeutic and organoleptic characteristics, diarylhepatanoids can be rightly termed as nutraceuticals. This review summarizes the wide range of pharmacological activities of diarylhepatanoids and nutraceutical formulations, with relevance to human health.


title: Common Scaffolds, Diverse Recognition Profiles

text: A paper in this issue of Structure reports the crystal structure of griffithsin, a lectin from red algae, and demonstrates its ability to bind and neutralize the SARS coronavirus, providing a link in understanding the evolution of lectins in this family. (Ziółkowska et al., 2006).


title: Discovery of Aptamers Targeting the Receptor-Binding Domain of the SARS-CoV-2 Spike Glycoprotein

text: [Image: see text] The World Health Organization has declared the outbreak of a novel coronavirus (SARS-CoV-2 or 2019-nCoV) as a global pandemic. However, the mechanisms behind the coronavirus infection are not yet fully understood, nor are there any targeted treatments or vaccines. In this study, we identified high-binding-affinity aptamers targeting SARS-CoV-2 RBD, using an ACE2 competition-based aptamer selection strategy and a machine learning screening algorithm. The K(d) values of the optimized CoV2-RBD-1C and CoV2-RBD-4C aptamers against RBD were 5.8 nM and 19.9 nM, respectively. Simulated interaction modeling, along with competitive experiments, suggests that two aptamers may have partially identical binding sites at ACE2 on SARS-CoV-2 RBD. These aptamers present an opportunity for generating new probes for recognition of SARS-CoV-2 and could provide assistance in the diagnosis and treatment of SARS-CoV-2 while providing a new tool for in-depth study of the mechanisms behind the coronavirus infection.


title: Les anti-inflammatoires et SARS-CoV-2 (COVID-19)

text: Résumé À ce jour, les options thérapeutiques pour le COVID-19 sévère restent limitées et il n’existe actuellement aucun médicament ni vaccin efficace pour le traitement ou la prévention du SARS-CoV-2 spécifique pour traiter les patients gravement malades. Une nouvelle stratégie de traitement, en plus de la thérapie antivirale seule, est susceptible d’être nécessaire pour avoir un impact significatif sur les résultats cliniques. Un traitement immunomodulateur pour réguler à la baisse de choc cytokinique peut fournir des informations sur le traitement de COVID-19. L’utilisation combinée d’un agent immunomodulateur avec un agent antiviral peut donner aux médecins plus de temps pour fournir un traitement de soutien aux patients atteints de COVID-19. Dans cette revue de la littérature, les rôles de plusieurs médicaments anti-inflammatoires ont été passés en revue pour explorer leur efficacité dans la lutte contre le SARS-CoV-2. Summary To date, treatment options for severe COVID-19 remain limited and there is currently no effective drug or vaccine for the treatment or prevention of specific SARS-CoV-2 to treat critically ill patients. A new treatment strategy, in addition to antiviral therapy alone, may be required to have a significant impact on clinical outcomes. Immunomodulatory therapy to downregulate cytokine shock may provide information on the treatment of COVID-19. Combined use of an immunomodulatory agent with an antiviral agent may give doctors more time to provide supportive therapy to patients with COVID-19. In this review of the literature, the roles of several anti-inflammatory drugs were reviewed to explore their effectiveness in the fight against SARS-CoV-2.


title: INFECTIOUS GASTROENTEROCOLITIDES IN CHILDREN An Update on Emerging Pathogens

text: The recognition that bacterial infections induce signal transduction responses in infected epithelial cells also provides new avenues to consider as novel forms of therapy. For example, the chemokine interleukin-8, which attracts neutrophils to sites of mucosal infection, is produced by epithelial cells of gastric and intestinal origin in response to bacterial infection. Inhibitors of chemokine production or inhibition of the biologic effects of neutrophil chemoattractants have the potential to reduce both mucosal inflammatory responses and the attendant clinical sequelae. Eukaryotic cells also respond to infection with elevations in cytosolic second messengers, including inositol triphosphate (IP3) and calcium ([Ca2+]i). In intestinal epithelium, these second messengers can mediate the diarrheal response to infection. Calcium/calmodulin inhibitors may have a beneficial effect in treating those gastrointestinal infections mediated through changes in the level of cytosolic free calcium. DuPont and colleagues showed, for example, that oral therapy with zaldaride maleate relieves symptoms of disease and shortens the duration of diarrhea in travelers with ETEC-induced diarrhea. Evaluation of additional signal transduction responses to microbial infections should provide both new insights into the pathogenesis of gastrointestinal infectious diseases and novel approaches to consider for the prevention and therapy for these human illnesses.


title: Programmed ribosomal frameshifting in HIV-1 and the SARS–CoV

text: Abstract Ribosomal frameshifting is a mechanism of gene expression used by several RNA viruses to express replicase enzymes. This article focuses on frameshifting in two human pathogens, the retrovirus human immunodeficiency virus type 1 (HIV-1) and the coronavirus responsible for severe acute respiratory syndrome (SARS). The nature of the frameshift signals of HIV-1 and the SARS–CoV will be described and the impact of this knowledge on models of frameshifting will be considered. The role of frameshifting in the replication cycle of the two pathogens and potential antiviral therapies targeting frameshifting will also be discussed.


title: Control measures to prevent Coronavirus disease 2019 （COVID‐19）pandemic in endoscopy centers: a multi‐centre study

text: AIMS: To investigate control measures for COVID‐19 pandemic in GIE centers in China. METHODS: This is a retrospective multi‐centre research, including seven centers. Data collection was from 1st Feb to 31st Mar 2020 and the same period last year. RESULTS: There were a total of 28 COVID‐19 definite cases in these hospitals. Six out of seven GIE centers were arranged to shut down on 1(st) Feb, with a mean shutdown days of 23.6±5.3. The actual workloads were only 10.3%‐62.9% compared to those last year. All centers had a preoperative COVID‐19 screening process. Epidemiological questionnaire, temperature taking and QR‐code of journey were conducted. Chest CT scan were conducted during the shutdown period and continued in five centers after return to work. Antibody and nucleic acid test were applied in 1‐3 centers. All endoscopists had advanced PPE. Five centers used surgical mask and the rest used N95 mask. Six centers used goggles or face shield. Five centers selected isolation gowns and the rest selected protective suits. The change frequency of these PPE was 4 hours. Sterilizing measures were improved in six centers. Five centers utilized ultraviolet and six centers strengthened natural ventilation. Four and six centers used peracetic acid during the period of shutdown and return to work, alone or matched with OPA or acidified water. CONCLUSIONS: Many effective control measures were conducted in GIE centers during the outbreak, including patients’ volume limitation, preoperative COVID‐19 screening, advanced PPE and disinfection methods.


title: Chapter 33 Molecular Mechanisms and Consequences of Immune and Nervous System Interactions

text: The nervous and immune systems are two complex physiologic systems that are functionally shaped by the integration of both genetic factors and experience. Although these systems are usually studied in isolation from each other, these two systems are highly interactive and cross-regulate the functions of each system. Therefore, disruptions in these normal neuroimmune interactions likely contribute to a broad array of neurologic and neurodegenerative disorders.


title: APOBEC3G directly binds Hepatitis B virus core protein in cell and cell free systems

text: Abstract APOBEC3G (A3G) is an intrinsic antiretroviral factor which can inhibit Hepatitis B virus (HBV) replication. This antiviral activity mainly depends on A3G incorporation into viral particles. However, the mechanisms of A3G packaging into HBV particles have not been well characterized. In this paper, we demonstrated that A3G interacted with the HBV core protein (HBc) directly in co-transfected HepG2 cells using the fluorescence resonance energy transfer (FRET) approach. In addition, we further found that this interaction did not require other factors in vitro using surface plasmon resonance (SPR) technology on BIAcore 3000. While cellular RNA or viral RNA was added to A3G protein solution before flow through the BIAcore chip, the interaction was not affected. In conclusion, these results suggest the possibility that A3G is incorporated into HBV viral particles via direct binding with HBc protein.


title: Expression of the Native Cholera Toxin B Subunit Gene and Assembly as Functional Oligomers in Transgenic Tobacco Chloroplasts

text: The B subunits of enterotoxigenic Escherichia coli (LTB) and cholera toxin of Vibrio cholerae (CTB) are candidate vaccine antigens. Integration of an unmodified CTB-coding sequence into chloroplast genomes (up to 10,000 copies per cell), resulted in the accumulation of up to 4.1% of total soluble tobacco leaf protein as functional oligomers (410-fold higher expression levels than that of the unmodified LTB gene expressed via the nuclear genome). However, expresssion levels reported are an underestimation of actual accumulation of CTB in transgenic chloroplasts, due to aggregation of the oligomeric forms in unboiled samples similar to the aggregation observed for purified bacterial antigen. PCR and Southern blot analyses confirmed stable integration of the CTB gene into the chloroplast genome. Western blot analysis showed that the chloroplast-synthesized CTB assembled into oligomers and were antigenically identical with purified native CTB. Also, binding assays confirmed that chloroplast- synthesized CTB binds to the intestinal membrane GM1-ganglioside receptor, indicating correct folding and disulfide bond formation of CTB pentamers within transgenic chloroplasts. In contrast to stunted nuclear transgenic plants, chloroplast transgenic plants were morphologically indistinguishable from untransformed plants, when CTB was constitutively expressed in chloroplasts. Introduced genes were inherited stably in subsequent generations, as confirmed by PCR and Southern blot analyses. Increased production of an efficient transmucosal carrier molecule and delivery system, like CTB, in transgenic chloroplasts makes plant-based oral vaccines and fusion proteins with CTB needing oral administration commercially feasible. Successful expression of foreign genes in transgenic chromoplasts and availability of marker-free chloroplast transformation techniques augurs well for development of vaccines in edible parts of transgenic plants. Furthermore, since the quaternary structure of many proteins is essential for their function, this investigation demonstrates the potential for other foreign multimeric proteins to be properly expressed and assembled in transgenic chloroplasts.


title: Unit-Based Staff Hand Hygiene (HH) Monitors To Improve Compliance in a Comprehensive Cancer Center

text: ISSUE: Both the Joint Commission on Accreditation of Healthcare Organization's (JCAHO) patient safety goal and the Centers for Disease Control and Prevention's (CDC) guidelines recommend monitoring of hand hygiene adherence and providing feedback to patient care staff. In order to advance patient safety and HH compliance, a three-pronged approach was implemented to enhance HH education, observation and measurement of HH compliance. PROJECT: Three components were implemented to improve compliance with HH practices: Education Component (Phase I) Development and implementation of a mandatory computer-based learning module on hand hygiene for all patient care staff that included HH's “Top Ten List” based on CDC's Guideline for Hand Hygiene in Health-Care Settings, 2002 Focused and targeted educational program for fellows, residents and mid-level providers including staff in invasive procedural areas Special educational HH program in Spanish for spanish-speaking staff Training of unit-based HH observers (100) using the Train-the-Trainer model Focus on HH compliance during Patient Safety Week Campaign in March 2005 Evaluation Component (Phase II) Three HH observational periods for assessment of HH compliance by trained HH monitors and infection control practitioners in all patient care areas Monthly self-assessment in patient care area units and diagnostic areas Communication and Feedback of Results Component (Phase III) Communication to staff, managers and administration regarding HH violations and compliance rates Prompt follow-up with re-education in patient care areas where HH violations rates were lower than expected. RESULTS: In Phase I, 100% of patient care staff complied with HH computer-based training. A total of 100 unit-based HH observers were trained using the Train-the-Trainer approach with education on HH basics, use of HH tool and feedback to colleagues and peers. In Phases II and III, observational periods with assessment of HH compliance were implemented by the trained HH observer of each clinical area. Data from observation periods, demonstrated an incremental improvement in HH practices with an initial baseline of 79% and an average of 91% in subsequent periods. LESSONS LEARNED: A coordinated systems approach that includes patient care staff, physicians, committees and administration is essential when initiating a process change. Providing education and feedback to staff and physicians will improve compliance. Identification of areas where additional education and feedback on HH is required and will also improve compliance.


title: Management of childhood-onset autoinflammatory diseases during the COVID-19 pandemic

text: Concerns regarding the comorbidity as a significant risk factor for Coronavirus Disease-2019 (COVID-19), gave rise to an urgent need for studies evaluating patients with chronic conditions such as autoinflammatory diseases (AIDs). We prepared a web-based survey investigating the clinical findings and contact histories among pediatric patients with AIDs. Confirmed COVID-19 cases, patients with contact history and those with symptoms which were highly suggestive of COVID-19 were called via phone or recruited to a video or face to face appointment. Data of AIDs were obtained from their medical records, retrospectively. Laboratory and screening findings were confirmed by our national health registry website. There were 404 patients (217 female) eligible for the enrollment. During pandemic, 375 (93%) were on colchicine treatment and 48 (11.8%) were receiving biologic treatment. Twenty-four out of 404 patients were admitted to hospital due to COVID-19 suspicion. Severe acute respiratory syndrome coronavirus-2 (SARS CoV-2) was identified through rhinopharyngeal swabs in seven patients, six of whom were only on colchicine treatment. Only one patient with no finding of any severe respiratory complications was hospitalized. All of seven patients recovered completely. Among patients on biologic drugs, neither a symptom nor a positive polymerase chain reaction test for COVID 19 was detected. In conclusion, pediatric patients with AIDs, those receiving biologic treatment and/or colchicine, may not be at increased risk for neither being infected nor the severe disease course.


title: Knowledge and Attitude of Dental Health Professionals about Middle East Respiratory Syndrome in Saudi Arabia

text: AIM AND OBJECTIVE: This study aims to evaluate the knowledge and attitude of practicing dental health professionals (DHPs) (dentist and dental auxiliaries) toward Middle East Respiratory Syndrome coronavirus (MERS-CoV) in Saudi Arabia. MATERIALS AND METHODS: A cross-sectional descriptive study was undertaken among practicing DHPs in Saudi Arabia. A total of 202 DHPs participated in this study. Knowledge and attitude were assessed using self-administered and pretested questionnaire. The questionnaire was administered online through Survey Monkey(®) program by sending link to the registered E-mail. Descriptive statistics were performed on demographic data. Mean knowledge and mean attitude scores of DHPs were calculated. Mann–Whitney U-test and Kruskal–Wallis tests were used to disclose the differences between study variables. Chi-square tests and Spearman's correlation tests were applied to find the associations between the variables. RESULTS: The study participants showed mean knowledge score of 12.26 ± 2.27 (based on 17 knowledge questions) and attitude score of 8.63 ± 1.68 (based on 10 attitude questions). The spearman's test showed the positive correlation between knowledge and attitude of DHPs about MERS (r = 0.093, P = 0.188). Knowledge gaps were reflected in questions related to the duration of infectivity (47.5%), treatment of MERS (39.6%), reservoir of MERS-CoV (38.1%), availability of vaccination against MERS-CoV (25.2%), the likelihood of infection (24.3%), and the type of MERS-CoV (23.3%). DHPs showed a positive attitude toward adherence to universal precautions given by CDC and WHO (0.94 ± 0.25), active participation infection control program (0.94 ± 0.24), and use of gowns, gloves, mask, and goggles while dealing with MERS-CoV patients (0.97 ± 0.17). Male DHPs showed significantly higher knowledge and positive attitude toward MERS-CoV infection compared to females. CONCLUSION: DHPs participated in this study showed good knowledge and positive attitude toward MERS. However, still few lacunae in the knowledge and attitudes toward MERS-CoV were found requiring extensive educational programs.


title: Ultraviolet Irradiation Enhances the Microbicidal Activity of Silver Nanoparticles by Hydroxyl Radicals

text: It is known that silver has microbicidal qualities; even at a low concentration, silver is active against many kinds of bacteria. Silver nanoparticles (AgNPs) have been extensively studied for a wide range of applications. Alternately, the toxicity of silver to human cells is considerably lower than that to bacteria. Recent studies have shown that AgNPs also have antiviral activity. We found that large amounts of hydroxyl radicals—highly reactive molecular species—are generated when AgNPs are irradiated with ultraviolet (UV) radiation with a wavelength of 365 nm, classified as ultraviolet A (UVA). In this study, we used electron spin resonance direct detection to confirm that UV irradiation of AgNPs produced rapid generation of hydroxyl radicals. As hydroxyl radicals are known to degrade bacteria, viruses, and some chemicals, the enhancement of the microbicidal activity of AgNPs by UV radiation could be valuable for the protection of healthcare workers and the prevention of the spread of infectious diseases.


title: Immunological environment shifts during pregnancy may affect the risk of developing severe complications in COVID‐19 patients

text: PROBLEM: The 2019 novel coronavirus disease (COVID‐19) has been rapidly spreading out in China and all over the world. Because of the immune reaction severity, 20.3% of COVID‐19 patients required intensive care unit, and 32.8% presented with acute respiratory distress syndrome (ARDS). In this regard, the urgent question that needs to be addressed promptly includes whether pregnant women with COVID‐19 will develop distinct symptoms from non‐pregnant and what are the reasons for that. RESULTS: According to the literature data, clinical characteristics of pregnant women with COVID‐19 were similar to those of non‐pregnant adults and no significant differences in gestational age, postpartum hemorrhage, and perineal resection rates between pregnant women with and without COVID‐19 were found. There is also a tendency that COVID‐19 disease progresses slowly in pregnant women and does not often result in fatal pregnancy complications. CONCLUSIONS: The predominant Th2‐type immunity and the action of T‐regs can play an important role in preventing the excess systemic inflammatory reaction and the development of life‐threatening complications as ARDS and MODS in COVID‐19 patients. This suggests a search for new drugs aimed at the suppression of effector functions and induction of tolerance, which will reduce the frequency of life‐threatening complications, including ARDS.


title: Fever of Unknown Origin Due to Zoonoses

text: Fevers of unknown origin have been classified as classic, nosocomial, immune-deficient, and HIV-related. More than half of the 1407 human pathogens are zoonotic, making zoonotic infections an important subcategory in each of the classifications. This article describes both common and unusual zoonoses causing fevers of unknown origin. Simian immune virus is considered as a possible emerging infection. For special populations (the homeless, zoophiliacs, those whose occupation or leisure brings them in close contact with oceans or lakes, and veterinarians), zoonotic infection potentials are discussed.


title: Knowledge, attitude and preventive practices related to COVID-19: a cross-sectional study in two Pakistani university populations

text: OBJECTIVE: The aim of this study was to assess COVID-19-related knowledge, attitudes, and preventive practices of two Pakistani university populations. METHODS: A cross-sectional study was conducted among students and employees of two higher education institutions in Lahore, Pakistan, namely the University of Lahore and the Gulab Devi Educational Complex. Participants were recruited using a convenient sampling method. A validated 45-item, self-administered questionnaire was used to collect data. Total possible scores were 0–14 for knowledge (scores < 7, 7–10 and > 10 were considered to indicate poor, moderate and good knowledge, respectively); 0–7 for attitude (scores > 5 were considered to indicate a good attitude); and 0–18 for COVID-19 preventative practices (scores > 12 were considered to indicate good preventative practices). RESULTS: Of the 417 enrolled participants, 416 reported that they were aware of COVID-19 and social media was the major source of their information. Mean scores were 10.12 ± 2.20 for knowledge (good, moderate and poor knowledge in 50.2%, 42.8% and 7.0% of participants, respectively); 5.74 ± 1.28 for attitude (65.4% of individuals had a positive attitude); and 11.04 ± 3.34 for COVID-19 preventative practices (only 36.5% of participants had good preventive practices). CONCLUSION: Pakistani university students and employees have good knowledge and attitudes regarding COVID-19, but unsatisfactory preventive practices. Therefore, health regulators should use multiple communication approaches, such as electronic, print and social media, phone messages, etc., to increase awareness and improve practices related to COVID-19.


title: An Overview of Current Approaches Toward the Treatment and Prevention of West Nile Virus Infection

text: The persistence of West Nile virus (WNV) infections throughout the USA since its inception in 1999 and its continuous spread throughout the globe calls for an urgent need of effective treatments and prevention measures. Although the licensing of several WNV vaccines for veterinary use provides a proof of concept, similar efforts on the development of an effective vaccine for humans remain still unsuccessful. Increased understanding of biology and pathogenesis of WNV together with recent technological advancements have raised hope that an effective WNV vaccine may be available in the near future. In addition, rapid progress in the structural and functional characterization of WNV and other flaviviral proteins have provided a solid base for the design and development of several classes of inhibitors as potential WNV therapeutics. Moreover, the therapeutic monoclonal antibodies demonstrate an excellent efficacy against WNV in animal models and represent a promising class of WNV therapeutics. However, there are some challenges as to the design and development of a safe and efficient WNV vaccine or therapeutic. In this chapter, we discuss the current approaches, progress, and challenges toward the development of WNV vaccines, therapeutic antibodies, and antiviral drugs.


title: Depression and Anxiety in Hong Kong during COVID-19

text: It has been three months since the first confirmed case of coronavirus disease 2019 (COVID-19) in Hong Kong, and people now have a more complete picture of the extent of the pandemic. Therefore, it is time to evaluate the impacts of COVID-19 on mental health. The current population-based study aimed to evaluate the depression and anxiety of people in Hong Kong during the COVID-19 pandemic. Respondents were randomly recruited and asked to complete a structured questionnaire, including the patient health questionnaire-9 (PHQ-9), the generalized anxiety disorder-7 (GAD-7), the global rating of change scale and items related to COVID-19. Of the 500 respondents included in the study, 19% had depression (PHQ-9 score ≥ 10) and 14% had anxiety (GAD score ≥ 10). In addition, 25.4% reported that their mental health had deteriorated since the pandemic. Multiple logistic regression analysis found that not experiencing the SARS outbreak in 2003, being worried about being infected by COVID-19, being bothered by having not enough surgical masks and being bothered by not being able to work from home were associated with a poorer mental health status. Psychological support, such as brief, home-based psychological interventions, should be provided to citizens during the pandemic.


title: An Animal Model of MERS Produced by Infection of Rhesus Macaques With MERS Coronavirus

text: In 2012, a novel coronavirus (CoV) associated with severe respiratory disease, Middle East respiratory syndrome (MERS-CoV; previously known as human coronavirus–Erasmus Medical Center or hCoV-EMC), emerged in the Arabian Peninsula. To date, 114 human cases of MERS-CoV have been reported, with 54 fatalities. Animal models for MERS-CoV infection of humans are needed to elucidate MERS pathogenesis and to develop vaccines and antivirals. In this study, we developed rhesus macaques as a model for MERS-CoV using intratracheal inoculation. The infected monkeys showed clinical signs of disease, virus replication, histological lesions, and neutralizing antibody production, indicating that this monkey model is suitable for studies of MERS-CoV infection.


title: Role of a habitat's air humidity in Covid-19 mortality

text: Abstract Transient local over-dry environment might be a contributor and an explanation for the observed asynchronous local rises in Covid-19 mortality. We propose that a habitat's air humidity negatively correlate with Covid-19 morbidity and mortality, and support this hypothesis on the example of publicly available data from German federal states.


title: Individualizing risk prediction for positive COVID-19 testing: results from 11,672 patients.

text: Abstract: Background Coronavirus disease-2019 (COVID-19) is sweeping the globe. Despite multiple case-series, actionable knowledge to proactively tailor decision-making is missing. Research Question Can a statistical model accurately predict infection with COVID? Study Design and Methods: We developed a prospective registry of all patients tested for COVID-19 in Cleveland Clinic to create individualized risk prediction models. We focus here on likelihood of a positive nasal or oropharyngeal COVID-19 test [COVID-19 (+)]. A least absolute shrinkage and selection operator (LASSO) logistic regression algorithm was constructed, which removed variables that were not contributing to the model’s cross-validated concordance index. Following external validation in a temporally and geographically-distinct cohort, the statistical prediction model was illustrated as a nomogram and deployed in an online risk calculator. Results 11,672 patients fulfilled study criteria in the development cohort, including 818 (7.0%) COVID-19 (+), and 2,295 patients fulfilled criteria in the validation cohort including 290 COVID-19 (+). Males, African Americans, older patients, and those with known COVID-19 exposure were at higher risk of being COVID-19 (+). Risk was reduced in those who had pneumococcal polysaccharide or influenza vaccine, or were on melatonin, paroxetine, or carvedilol. Our model had favorable discrimination (c-statistic=0.863 in development; 0.840 in validation cohort) and calibration. We present sensitivity, specificity, negative predictive value, and positive predictive value at different prediction cut-offs.The calculator is freely available at https://riskcalc.org/COVID19. Interpretation Prediction of a COVID-19 (+) test is possible and could help direct healthcare resources. We demonstrate relevance of age, race, gender, and socioeconomic characteristics in COVID-19-susceptibility and suggest a potential modifying role of certain common vaccinations and drugs identified in drug-repurposing studies.


title: Canine Monocytic Ehrlichiosis among working dogs of organised kennels in India: A comprehensive analyses of clinico-pathology, serological and molecular epidemiological approach

text: Abstract Canine Monocytic Ehrlichiosis (CME) is a serious tick-borne rickettsial disease affecting canine populations globally. Besides few reports from stray and pet dogs from localised geographical regions (cities/towns/small states), a comprehensive study on prevalence of Ehrlichia canis (E. canis) among working dogs from different geo-climatic zones of India was pertinently lacking. Study of CME among these dog populations was thus carried out, encompassing clinical aspects and different diagnostic methodologies viz., microscopy, serology and molecular biology. During the two-year study period, clinical specimens from 225 cases suspected of canine ehrlichiosis were examined for clinical pathology and presence of the haemoparasites. Overall prevalence of ehrlichiosis by microscopic examination, commercial dot-ELISA kit and nested PCR assay was estimated to be 1.3%, 19.1% and 5.8%, respectively, which were found to be statistically significant by McNemar Chi squared test (p<0.05). It was also observed that possibly due to widespread use of doxycycline therapy in field, CME presently does not remain a potential threat which it uses to pose earlier. However, concurrent infections of E. canis and Babesia gibsoni were found to be mostly fatal. Keeping in view of high number of apparently healthy dogs (24) out of total positive cases (46) observed during the study, it is recommended that prevalence studies on CME should also involve screening of apparently healthy dogs. Phylogenetic analysis carried on partial sequencing of 16S rRNA of E. canis strains revealed that all of the Indian strains clustered in a single clade with other E. canis species from India and rest of the world. Molecular divergence was observed among the sequences of Brazilian and American isolates which were also included in the present study. These findings have thus opened a new paradigm for planning of pragmatic control strategies against CME.


title: Transcriptomic Characterization of the Novel Avian-Origin Influenza A (H7N9) Virus: Specific Host Response and Responses Intermediate between Avian (H5N1 and H7N7) and Human (H3N2) Viruses and Implications for Treatment Options

text: A novel avian-origin H7N9 influenza A virus (IAV) emerged in China in 2013, causing mild to lethal human respiratory infections. H7N9 originated with multiple reassortment events between avian viruses and carries genetic markers of human adaptation. Determining whether H7N9 induces a host response closer to that with human or avian IAV is important in order to better characterize this emerging virus. Here we compared the human lung epithelial cell response to infection with A/Anhui/01/13 (H7N9) or highly pathogenic avian-origin H5N1, H7N7, or human seasonal H3N2 IAV. The transcriptomic response to H7N9 was highly specific to this strain but was more similar to the response to human H3N2 than to that to other avian IAVs. H7N9 and H3N2 both elicited responses related to eicosanoid signaling and chromatin modification, whereas H7N9 specifically induced genes regulating the cell cycle and transcription. Among avian IAVs, the response to H7N9 was closest to that elicited by H5N1 virus. Host responses common to H7N9 and the other avian viruses included the lack of induction of the antigen presentation pathway and reduced proinflammatory cytokine induction compared to that with H3N2. Repression of these responses could have an important impact on the immunogenicity and virulence of H7N9 in humans. Finally, using a genome-based drug repurposing approach, we identified several drugs predicted to regulate the host response to H7N9 that may act as potential antivirals, including several kinase inhibitors, as well as FDA-approved drugs, such as troglitazone and minocycline. Importantly, we validated that minocycline inhibited H7N9 replication in vitro, suggesting that our computational approach holds promise for identifying novel antivirals.


title: Fluorescence in situ hybridization investigation of potentially pathogenic bacteria involved in neonatal porcine diarrhea

text: BACKGROUND: Neonatal diarrhea is a multifactorial condition commonly present on pig farms and leads to economic losses due to increased morbidity and mortality of piglets. Immature immune system and lack of fully established microbiota at birth predispose neonatal piglets to infection with enteric pathogens. The microorganisms that for decades have been associated with enteritis and diarrhea in suckling piglets are: rotavirus A, coronavirus, enterotoxigenic Escherichia coli (ETEC), Clostridium perfringens type C, Cryptosporidium spp., Giardia spp., Cystoisospora suis and Strongyloides ransomi. However, in recent years, the pig industry has experienced an increased number of neonatal diarrhea cases in which the above mentioned pathogens are no longer detected. Potentially pathogenic bacteria have recently received focus in the research on the possible etiology of neonatal diarrhea not caused by common pathogens. The primary aim of this study was to investigate the role of E. coli, Enterococcus spp., C. perfringens and C. difficile in the pathogenesis of neonatal porcine diarrhea with no established casual agents. Fluorescence in situ hybridization with oligonucleotide probes was applied on the fixed intestinal tissue samples from 51 diarrheic and 50 non-diarrheic piglets collected from four Danish farms during outbreaks of neonatal diarrhea not caused by well-known enteric pathogens. Furthermore, an association between the presence of these bacteria and histological lesions was evaluated. RESULTS: The prevalence of fluorescence signals specific for E. coli, C. perfringens and C. difficile was similar in both groups of piglets. However, Enterococcus spp. was primarily detected in the diarrheic piglets. Furthermore, adherent bacteria were detected in 37 % diarrheic and 14 % non-diarrheic piglets. These bacteria were identified as E. coli and Enterococcus spp. and their presence in the intestinal mucosa was associated with histopathological changes. CONCLUSIONS: The results of this study showed that simultaneous colonization of the intestinal mucosa by adherent non-ETEC E. coli and Enterococcus spp. can be involved in the pathogenesis of neonatal porcine diarrhea. These bacteria should be considered in diagnosis of diarrhea in piglets, when detection of common, well-known enteric agents is unsuccessful.


title: Selecting a SARS-CoV-2/COVID molecular testing method for your laboratory

text: In early March 2020 it became apparent that clinical laboratories would need to quickly develop strategies for SARS-CoV-2/COVID-19 testing. For most, the initial approach was to send out testing to a reference laboratory. As the pandemic has progressed, the food and drug administration (FDA) has allowed for several manufacturers to make testing reagents commercially available. Concurrently, the demand for rapid accessibility of results persists, leading many laboratories to evaluate options for “in house” testing. This reflection highlights some of the considerations when selecting the best method for your laboratory, with specific examples highlighted from a medium volume laboratory’s experience.


title: Risk factors for disease progression in hospitalized patients with COVID-19: a retrospective cohort study

text: Background: To investigate the risk factors related to aggravation and clinical outcomes in coronavirus disease 2019 (COVID-19) patients. Methods: We performed a retrospective study on the risk factors for disease progression of cases with COVID-19. Based on the clinical types, the patients were divided into a progression group and an improvement group. Multivariable logistic regression and ROC curve analysis were performed to explore the risk factors for disease progression. Results: A total of 101 patients were included in this study; diseases progression occurred in 17 patients, 84 patients improved, 6 were transferred to intensive care unit (ICU), and 5 died. The mean time to obtain negative nucleic acid results was 12.5 ± 5.0 days. Multivariate logistic analysis indicated that age (OR, 0.104; p = .002), C-reactive protein (CRP) (OR, 0.093; p < .001) and lymphocyte count (OR, 3.397; p = .022) were risk factors for disease progression. ROC curve analysis revealed that the AUC of age, CRP and lymphocyte count for disease progression were 0.873, 0.911 and 0.817, respectively. Conclusions: Older age increased CRP and decreased lymphocyte count resulted in potential risk factors for COVID-19 progression. This may be helpful in identifying patients whose condition worsens at an early stage.


title: An Evaluation of the Additive Effect of Natural Herbal Medicine on SARS or SARS-like Infectious Diseases in 2003: A Randomized, Double-blind, and Controlled Pilot Study

text: Natural herbal medicine (NHM) has been used to control infectious diseases for thousands of years. In view of the possible beneficial effect of NHM on SARS, we conducted this study to examine whether NHM is of any benefit as a supplementary treatment of SARS or SARS-like infectious disease. This was a randomized, double-blind, placebo-controlled trial. Twenty-eight patients fulfilled the WHO inclusion criteria and our exclusion criteria. All enrolled patients received routine western-medicine treatment. Patients were randomly allocated to one of the three supplementary treatment groups: NHM A (Group A, n = 9) NHM B (Group B, n = 9) or placebo (Group C, n = 10). Chest X-ray was done every 1 or 2 days for every patient. Reading radiologists use a standard 0–3 scoring system (0: no infiltration; 1: focal haziness or even small patchy lesion; 2: ground glass picture; 3: lobar consolidation) according to the severity of infiltration in each lung field (three lung fields in both right and left lungs). The main outcome measurements were the improving chest radiographic scores (IRS) and the duration (days) till improvement (DI). One patient from the placebo group passed away. Patients from NHM A took less days before showing improvement (6.7 ± 1.8) compared with placebo group (11.2 ± 4.9), which showed statistical significance (P = 0.04). The cases were too few to be conclusive, the initial observations seem to indicate NHM appears to be safe in non-criticallly ill patients and clinical trials are feasible in the setting of pandemic outbreaks.


title: Early detection of antibodies against various structural proteins of the SARS-associated coronavirus in SARS patients

text: Severe acute respiratory syndrome (SARS), a new disease with symptoms similar to those of atypical pneumonia, raised a global alert in March 2003. Because of its relatively high transmissibility and mortality upon infection, probable SARS patients were quarantined and treated with special and intensive care. Therefore, instant and accurate laboratory confirmation of SARS-associated coronavirus (SARS-CoV) infection has become a worldwide interest. For this need, we purified recombinant proteins including the nucleocapsid (N), envelope (E), membrane (M), and truncated forms of the spike protein (S1–S7) of SARS-CoV inEscherichia coli. The six proteins N, E, M, S2, S5, and S6 were used for Western blotting (WB) to detect various immunoglobulin classes in 90 serum samples from 54 probable SARS patients. The results indicated that N was recognized in most of the sera. In some cases, S6 could be recognized as early as 2 or 3 days after illness onset, while S5 was recognized at a later stage. Furthermore, the result of recombinant-protein-based WB showed a 90% agreement with that of the whole-virus-based immunofluorescence assay. Combining WB with existing RT-PCR, the laboratory confirmation for SARS-CoV infection was greatly enhanced by 24.1%, from 48.1% (RT-PCR alone) to 72.2%. Finally, our results show that IgA antibodies against SARS-CoV can be detected within 1 week after illness onset in a few SARS patients.