title: Detection of rhinovirus RNA in nasal epithelial cells by in situ hybridization

text: Abstract This paper describes the development and evaluation of in situ hybridization (ISH) for the detection of rhinovirus in cells obtained from nasal washings of volunteers infected with human rhinovirus 14 (HRV-14). Twenty-five (66%) and 27 (71%) of 38 volunteers inoculated with HRV-14 had evidence of infection by virus isolation and ISH, respectively, on at least one of 4 days investigated after virus challenge. In contrast, only 14 of 38 (37%) volunteers had significant antibody rises as detected by the neutralization test. Of the 38 volunteers inoculated with HRV-14, only 13 (34%) had symptoms of colds. Of these, 12 (92%) and 10 (77%) were positive by virus isolation or ISH, respectively, on at least one day. Six (46%) had significant antibody rises by neutralization. Similarly, of the 38 volunteers challenged, 22 (58%) were asymptomatic and of these 10 (45.5%) and 12 (54.5%) were positive by virus isolation and ISH, respectively, on at least one day. Only 8 (36.4%) of these asymptomatic volunteers showed significant antibody rises by neutralization. There were significant associations between the detection of rhinoviruses by ISH and virus isolation on the third day (P<0.025) after virus challenge in the group as a whole and in the symptomatic group. These results show that generally rhinovirus detection by ISH compares well with virus isolation and both tests are clearly more sensitive than the neutralization test in detecting evidence of infection. It is concluded that ISH is an interesting new technique that may play an important role in the study of rhinovirus infection and pathogenesis.


title: Harnessing DNA Synthesis to Develop Rapid Responses to Emerging and Pandemic Pathogens

text: Given the interconnected nature of our world today, emerging pathogens and pandemic outbreaks are an ever-growing threat to the health and economic stability of the global community. This is evident by the recent 2009 Influenza A (H1N1) pandemic, the SARS outbreak, as well as the ever-present threat of global bioterrorism. Fortunately, the biomedical community has been able to rapidly generate sequence data so these pathogens can be readily identified. To date, however, the utilization of this sequence data to rapidly produce relevant experimental results or actionable treatments is lagging in spite of obtained sequence data. Thus, a pathogenic threat that has emerged and/or developed into a pandemic can be rapidly identified; however, translating this identification into a targeted therapeutic or treatment that is rapidly available has not yet materialized. This commentary suggests that the growing technology of DNA synthesis should be fully implemented as a means to rapidly generate in vivo data and possibly actionable therapeutics soon after sequence data becomes available.


title: Snapshot of Viral Infections in Wild Carnivores Reveals Ubiquity of Parvovirus and Susceptibility of Egyptian Mongoose to Feline Panleukopenia Virus

text: The exposure of wild carnivores to viral pathogens, with emphasis on parvovirus (CPV/FPLV), was assessed based on the molecular screening of tissue samples from 128 hunted or accidentally road-killed animals collected in Portugal from 2008 to 2011, including Egyptian mongoose (Herpestes ichneumon, n = 99), red fox (Vulpes vulpes, n = 19), stone marten (Martes foina, n = 3), common genet (Genetta genetta, n = 3) and Eurasian badger (Meles meles, n = 4). A high prevalence of parvovirus DNA (63%) was detected among all surveyed species, particularly in mongooses (58%) and red foxes (79%), along with the presence of CPV/FPLV circulating antibodies that were identified in 90% of a subset of parvovirus-DNA positive samples. Most specimens were extensively autolysed, restricting macro and microscopic investigations for lesion evaluation. Whenever possible to examine, signs of active disease were not present, supporting the hypothesis that the parvovirus vp2 gene fragments detected by real-time PCR possibly correspond to viral DNA reminiscent from previous infections. The molecular characterization of viruses, based on the analysis of the complete or partial sequence of the vp2 gene, allowed typifying three viral strains of mongoose and four red fox’s as feline panleukopenia virus (FPLV) and one stone marten’s as newCPV-2b type. The genetic similarity found between the FPLV viruses from free-ranging and captive wild species originated in Portugal and publicly available comparable sequences, suggests a closer genetic relatedness among FPLV circulating in Portugal. Although the clinical and epidemiological significance of infection could not be established, this study evidences that exposure of sympatric wild carnivores to parvovirus is common and geographically widespread, potentially carrying a risk to susceptible populations at the wildlife-domestic interface and to threatened species, such as the wildcat (Felis silvestris) and the critically endangered Iberian lynx (Lynx pardinus).


title: Epidemiology of Viral Pneumonia

text: The burden of pneumonia, including that due to respiratory viruses, is markedly higher in the very young (<5 years) and older adults (≥50 years). Respiratory viruses substantially contribute to pneumonia in both adults and children, and when systematically tested for, are more commonly detected than bacteria in both adults and children. It is difficult to distinguish between viruses by clinical presentation, and the exact clinical implication of viral detections among patients with pneumonia depends on the pathogen detected; however, there is increasing evidence of their importance in pneumonia.


title: Telemental health comes into its own with social distancing

text: Telepsychiatry, telemental health, telepsychology, teletherapy — whatever you call it, it is now required in the era of the coronavirus/COVID‐19 pandemic. “We should be doing it immediately; the more, the better,” said Robert Caudill, M.D., a member of the American Psychiatric Association's Committee on Telepsychiatry. “It's exploded in the past week, obviously,” he told CABL in a Zoom interview on March 23. But he thinks it should be the norm.


title: Production of second-generation cloned cats by somatic cell nuclear transfer

text: Abstract We successfully produced second-generation cloned cats by somatic cell nuclear transfer (SCNT) using skin cells from a cloned cat. Skin cells from an odd-eyed, all-white male cat (G0 donor cat) were used to generate a cloned cat (G1 cloned cat). At 6 months of age, skin cells from the G1 cloned cat were used for SCNT to produce second-generation cloned cats. We compared the in vitro and in vivo development of SCNT embryos that were derived from the G0 donor and G1 cloned donor cat's skin fibroblasts. The nuclei from the G0 donor and G1 cloned donor cat's skin fibroblasts fused with enucleated oocytes with equal rates of fusion (60.7% vs. 58.8%, respectively) and cleavage (66.3% vs. 63.4%). The 2–4-cell SCNT embryos were then transferred into recipients. One of the five recipients of G0 donor derived NT embryos (20%) delivered one live male cloned kitten, whereas 4 of 15 recipients of the G1 cloned donor cat derived NT embryos (26%) delivered a total of seven male second-generation cloned kittens (four live kittens from one surrogate, plus two stillborn kittens, and one live kitten that died 2d after birth from three other surrogate mothers). The four second-generation cloned kittens from the same surrogate all had a white coat color; three of the four second-generation cloned kittens had two blue eyes, and one of the second-generation cloned kittens had an odd-eye color. Despite low cloning efficiency, cloned cats can be used as donor cats to produce second-generation cloned cats.


Positive Screening for Wuhan Novel Coronavirus Infection at International Airport: What's the Final Diagnosis for Positive Cases


title: The contribution of dry indoor built environment on the spread of Coronavirus: Data from various Indian states

text: Coronavirus spread is more serious in urban metropolitan cities compared to rural areas. It is observed from the data on the infection rate available in the various sources that the cold and dry conditions accelerate the spread of coronavirus. In the present work, the existing theory of respiratory droplet drying is used to propose the mechanism of virus spread under various climates and the indoor environment conditions which plays a greater role in the virus spread. This concept is assessed using four major parameters such as population density, climate severity, the volume of indoor spaces, and air-conditioning usage which affect the infection spread and mortality using the data available for various states of India. Further, it is analysed using the data from various states in India along with the respective climatic conditions. It is found that under some indoor scenarios, the coronaviruses present in the respiratory droplets become active due to size reduction that occurs both in sessile and airborne droplet nuclei causing an increase in the spread. Understanding this mechanism will be very useful to take the necessary steps to reduce the rate of transmission by initiating corrective measures and maintaining the required conditions in the indoor built environment.


Recommended Practices for Prevention of Transmissible Infections in the Perioperative Practice Setting


title: Laboratory Diagnosis of SARS

text: The virologic test results of 415 patients with severe acute respiratory syndrome (SARS) were examined. The peak detection rate for SARS-associated coronavirus occurred at week 2 after illness onset for respiratory specimens, at weeks 2 to 3 for stool or rectal swab specimens, and at week 4 for urine specimens. The latest stool sample that was positive by reverse transcription–polymerase chain reaction (RT-PCR) was collected on day 75 while the patient was receiving intensive care. Tracheal aspirate and stool samples had a higher diagnostic yield (RT-PCR average positive rate for first 2 weeks: 66.7% and 56.5%, respectively). Pooled throat and nasal swabs, rectal swab, nasal swab, throat swab, and nasopharyngeal aspirate specimens provided a moderate yield (29.7%–40.0%), whereas throat washing and urine specimens showed a lower yield (17.3% and 4.5%). The collection procedures for stool and pooled nasal and throat swab specimens were the least likely to transmit infection, and the combination gave the highest yield for coronavirus detection by RT-PCR. Positive virologic test results in patient groups were associated with mechanical ventilation or death (p < 0.001), suggesting a correlation between viral load and disease severity.


title: Structures and Polymorphic Interactions of Two Heptad-Repeat Regions of the SARS Virus S2 Protein

text: Summary Entry of SARS coronavirus into its target cell requires large-scale structural transitions in the viral spike (S) glycoprotein in order to induce fusion of the virus and cell membranes. Here we describe the identification and crystal structures of four distinct α-helical domains derived from the highly conserved heptad-repeat (HR) regions of the S2 fusion subunit. The four domains are an antiparallel four-stranded coiled coil, a parallel trimeric coiled coil, a four-helix bundle, and a six-helix bundle that is likely the final fusogenic form of the protein. When considered together, the structural and thermodynamic features of the four domains suggest a possible mechanism whereby the HR regions, initially sequestered in the native S glycoprotein spike, are released and refold sequentially to promote membrane fusion. Our results provide a structural framework for understanding the control of membrane fusion and should guide efforts to intervene in the SARS coronavirus entry process.


title: Pathogen-specific risk factors in acute outbreaks of respiratory disease in calves

text: ABSTRACT Respiratory tract infections (bovine respiratory disease) are a major concern in calf rearing. The objective of this study was to identify pathogen-specific risk factors associated with epidemic respiratory disease in calves. A cross-sectional study was conducted, involving 128 outbreaks (29 dairy, 58 dairy-mixed, and 41 beef) in Belgium (2016–2018). A semiquantitative PCR for 7 respiratory pathogens was done on a pooled nonendoscopic bronchoalveolar lavage sample for each herd. Potential risk factors were collected by questionnaire and derived from the national cattle registration databank. Most outbreaks occurred between October and March, and single and multiple viral infections were detected in 58.6% (75/128) and 13.3% (17/128), respectively. Bovine coronavirus (BCV) was the most frequently isolated virus (38.4%), followed by bovine respiratory syncytial virus (bRSV; 29.4%) and parainfluenzavirus type 3 (PI-3; 8.1%). Mycoplasma bovis, Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni were detected in 33.3, 41.2, 89.1, and 36.4% of the herds, respectively. Specific risk factors for BCV detection were detection of M. haemolytica [odds ratio (OR) = 2.8 (95% confidence interval = 1.1–7.5)], increasing herd size [OR = 1.3 (1.0–1.8) for each increase with 100 animals] and detection of BCV by antigen ELISA on feces in calves in the last year [OR = 3.6 (1.2–11.1)]. A seasonal effect was shown for bRSV only {more in winter compared with autumn [OR = 10.3 (2.8–37.5)]}. Other factors associated with bRSV were PI-3 detection [OR = 13.4 (2.1–86.0)], prevalence of calves with respiratory disease [OR = 1.02 (1.00–1.04) per 1% increase], and number of days with respiratory signs before sampling [OR = 0.99 (0.98–0.99) per day increase]. Next to its association with BCV, M. haemolytica was more frequently detected in herds with 5 to 10 animals per pen [OR = 8.0 (1.4–46.9)] compared with <5 animals, and in herds with sawdust as bedding [OR = 18.3 (1.8–191.6)]. Also, for H. somni, housing on sawdust was a risk factor [OR = 5.2 (1.2–23.0)]. Purchase of cattle [OR = 2.9 (1.0–8.0)] and housing of recently purchased animals in the same airspace [OR = 5.0 (1.5–16.5)] were risk factors for M. bovis. This study identified pathogen-specific risk factors that might be useful for the development of customized control and prevention and for the design of decision support tools to justify antimicrobial use by predicting the most likely pathogen before sampling results are available.


Novel Middle East respiratory syndrome coronavirus


title: Indirubin, a bisindole alkaloid from Isatis indigotica, reduces H1N1 susceptibility in stressed mice by regulating MAVS signaling

text: Isatis indigotica has a long history in treating virus infection and related symptoms in China. Nevertheless, its antivirus evidence in animal studies is not satisfactory, which might be due to the lack of appropriate animal model. Previously, we had utilized restraint stress to establish mouse H1N1 susceptibility model which was helpful in evaluating the anti-virus effect of medicines targeting host factors, such as type I interferon production. In this study, this model was employed to investigate the effect and mechanism of indirubin, a natural bisindole alkaloid from Isatis indigotica, on influenza A virus susceptibility. In the in vitro study, the stress hormone corticosterone was used to simulate restraint stress. Our results demonstrated that indirubin decreased the susceptibility to influenza virus with lowered mortality and alleviated lung damage in restraint-stressed mice model. Moreover, indirubin promoted the expression of interferon-β and interferon inducible transmembrane 3. In addition, indirubin maintained the morphology and function of mitochondria following influenza A virus infection. Further study revealed that indirubin promoted interferon-β production through promoting mitochondrial antiviral signaling pathway. Our study indicated that indirubin could be a candidate for the therapy of influenza.


title: Genotypic analysis of virulence genes and antimicrobial profile of diarrheagenic Escherichia coli isolated from diseased lambs in Iran

text: The aim of the present study was to determine the analysis of virulence genes and antimicrobial profile of diarrheagenic Escherichia coli isolated from diseased lambs. Two hundred ninety E. coli isolates were recovered from 300 rectal swabs of diarrheic lambs and were confirmed by biochemical tests. The pathotype determination was done according to the presence of genes including f5, f41, LTI, STI, bfp, ipaH, stx (1), stx (2), eae, ehlyA, cnf (1), cnf (2), cdIII, cdIV, and f17 by PCR method. Sixty-six isolates (23.72%) possessed the STI gene and categorized into entrotoxigenic E. coli (ETEC). Nine isolates (3.1%) and five isolates (1.72%) were positive for the cnf1 and cnf2 genes which categorized into necrotoxic E. coli (NTEC). Hundred and seventeen isolates (40.34%) harbored stx (1) and/or stx (2) and classified as Shiga toxin-producing E. coli (STEC). Thirteen isolates (4.48%) were assigned to atypical entropathogenic E. coli (aEPEC) and possessed eae gene. Two isolates (0.68%) were positive for ipaH gene and were assigned to entroinvasive E. coli (EIEC). Statistical analysis showed a specific association between eae gene and STEC pathotype (P < 0.0001). The most prevalent resistance was observed against lincomycin (96.5%) and the lowest resistance was against kanamycine (56.02%), respectively. The high prevalence of STEC and ETEC indicates that diarrheic lambs represent an important reservoir for humans. ETEC may play an important role for frequent occurrence of diarrhea in lambs observed in this region. Due to high antibiotic resistance, appropriate control should be implemented in veterinary medicine to curb the development of novel resistant isolates.


title: Nontraditional Antibiotics—Challenges and Triumphs

text: The pursuit of nontraditional antibiotics is becoming an increasingly important means to tackle seemingly insurmountable challenges faced by contemporary antibiotic researchers as they overcome the shifting landscape of bacterial pathogenesis, particularly for Gram-negative bacteria [...].


157 Coronaviruses, Including Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS)


title: The demand for inpatient and ICU beds for COVID-19 in the US: lessons from Chinese cities

text: BACKGROUND: Sustained spread of SARS-CoV-2 has happened in major US cities. Capacity needs in Chinese cities could inform the planning of local healthcare resources. METHODS: We described the intensive care unit (ICU) and inpatient bed needs for confirmed COVID-19 patients in two Chinese cities (Wuhan and Guangzhou) from January 10 to February 29, 2020, and compared the timing of disease control measures in relation to the timing of SARS-CoV-2 community spread. We estimated the peak ICU bed needs in US cities if a Wuhan-like outbreak occurs. RESULTS: In Wuhan, strict disease control measures were implemented six weeks after sustained local transmission of SARS-CoV-2. Between January 10 and February 29, COVID-19 patients accounted for an average of 637 ICU patients and 3,454 serious inpatients on each day. During the epidemic peak, 19,425 patients (24.5 per 10,000 adults) were hospitalized, 9,689 (12.2 per 10,000 adults) were considered to be in serious condition, and 2,087 patients (2.6 per 10,000 adults) needed critical care per day. In Guangzhou, strict disease control measures were implemented within one week of case importation. Between January 24 and February 29, COVID-19 accounted for an average of 9 ICU patients and 20 inpatients on each day. During the epidemic peak, 15 patients were in critical condition, and 38 were classified as serious. If a Wuhan-like outbreak were to happen in a US city, the need for healthcare resources may be higher in cities with a higher prevalence of vulnerable populations. CONCLUSION: Even after the lockdown of Wuhan on January 23, the number of seriously ill COVID-19 patients continued to rise, exceeding local hospitalization and ICU capacities for at least a month. Plans are urgently needed to mitigate the effect of COVID-19 outbreaks on the local healthcare system in US cities.


title: Gastroenteritis, Viral

text: Abstract Viral gastroenteritis (VGE) is an acute, self-limiting gastrointestinal infection caused by several viruses which most often trigger nausea, vomiting, abdominal pain and diarrhea. It is one of the most common human illnesses with its greatest impact on the very young and elderly. In the developing world, VGE can be life threatening whereas in most developed countries, it is self-limited and managed with hydration. Most agents of VGE are spread by the fecal oral route or may occur in explosive outbreaks due to food or water contamination. Control of these infections depends upon proper food handling, hand hygiene, clean water and the rotavirus vaccination of infants. This chapter will discuss the common viruses which cause this syndrome.


title: Molecular detection and genetic characterization of kobuviruses and astroviruses in asymptomatic local pigs in East Africa

text: In this study, swine fecal specimens (n = 251) collected from nursing and weaned piglets raised under smallholder production systems were screened for the presence of kobuviruses by RT-PCR. Porcine kobuviruses were detected in 13.1 % (33/251) of the samples. We demonstrated that porcine kobuvirus infections exist in indigenous pigs in Kenya and Uganda and that the prevalence was higher in young piglets than older pigs: nursing piglets (15 %), post-weaning (3-month-old) pigs (17 %), 4-month-old pigs (10 %). Genetic analysis of the partial RNA-dependent RNA polymerase (RdRp) region (690 nt) revealed that kobuviruses circulating in East Africa are diverse, sharing nucleotide sequence identities ranging from 89.7 to 99.1 % and 88 to 92.3 % among them and with known porcine kobuviruses, respectively. The nucleotide sequence identities between our kobuvirus strains and those of human, bovine and canine kobuviruses were 69.4-70.7 %, 73.1-74.4 % and 67-70.7 %, respectively. Additionally, upon sequencing selected samples that showed consistent 720-bp RT-PCR bands while using the same primer set, we detected porcine astroviruses in our samples belonging to type 2 and type 3 mamastroviruses. To our knowledge, this study reports the first detection and molecular analysis of both porcine kobuviruses and astroviruses in an African region. Further studies are required to determine the role of these viruses in gastrointestinal infections of pigs in this region and to determine the genetic diversity of the circulating strains to develop accurate diagnostic tools and implement appropriate control strategies.


Notes on Covid-19 and the contradictions of capital: a call for papers


title: Surface-displayed porcine epidemic diarrhea viral (PEDV) antigens on lactic acid bacteria

text: In this report, for surface display of viral antigen on lactobacilli, we have developed a surface antigen display system using the poly-γ-glutamate synthetase A protein (pgsA) of Bacillus subtilis as an anchoring matrix. Recombinant fusion proteins comprised of pgsA and neucleocapsid protein of PEDV were stably expressed in Lactobacillus casei. Surface location of fusion protein was verified by ELISA, immunofluoresence microscopy. Oral and intranasal inoculations of recombinant L. casei into pregnant sow and mice resulted in high levels of serum immunoglobuline G (IgG) and mucosal IgA, as demonstrated by rnELISA(recombinant N protein ELISA) using recombinant N protein. Absorbance of IgG in pregnant sow sera highly increased duration of the experiment. More importantly, the level of IgA in colostrum were increased significantly higher than that of IgG. The IgG levels of the piglets were increased after suckling colostrum secreted from sows previously inoculated recombinant L. casei. These results indicate that mucosal immunization with recombinant L. casei expressing PEDV N protein (neucleoprotein of PEDV) on its surface elicited high levels of mucosal IgA and circulation IgG immune responses against the antigen N of PEDV.


title: Detection of serum neopterin for early assessment of dengue virus infection

text: Summary Objective Neopterin is generated and released in increased amounts by macrophages upon activation by interferon-γ during Th1-type immune response. The potential usefulness of neopterin in early prognostic information of dengue virus infection was investigated. Methods Neopterin concentrations were determined in serum samples from 110 dengue fever (DF) patients. The neopterin levels were compared with those in 50 measles and 40 influenza patients; 155 healthy blood donors served as controls. Results In acute sera of DF patients mean neopterin concentration was 48.2nmol/L, which was higher than that in patients with measles (mean: 36.3nmol/L) and influenza (18.8nmol/L) and in healthy controls (6.7nmol/L; P <0.001). In the patients with confirmed DF, an early neopterin elevation was detected already at the first day after the onset of symptoms and rose to a maximum level of 54.3nmol/L 4days after the onset. Higher increase of neopterin level in DF patients was associated with longer duration of fever and thus predicted the clinical course of the disease. Conclusions Neopterin concentrations were found significantly higher in DF patients compared with healthy controls and also with other viral infections (P <0.001) and may allow early assessment of the severity of DF.


title: Virulent feline calicivirus disease in a shelter in italy: A case description

text: Abstract Feline calicivirus (FCV) is a common pathogen of cats that is particularly widespread in multi-cat environments such as shelters and catteries. FCV infections are usually associated with acute, mild and self-limiting upper respiratory tract disease characterized by oral vesicles/ulcers. Recently, virulent systemic disease (VSD) associated with FCV infection has been reported in the USA and Europe. This paper describes a case of VSD affecting one adult, FIV infected cat (“Oscar”) living in a shelter located in Northern Italy; the clinical, post-mortem and laboratory findings indicate that this is the first case of suspected FCV-VSD in this country. Similar to a previous report (Meyer et al., 2011), the disease affected only one cat, while others remained asymptomatic, despite their direct contact with “Oscar”. Phylogenetic analysis identified unique features in the “Oscar” FCV isolate. The FIV infection of the patient might have favoured the generation of the virulent FCV strains in this cat.


Three-Dimensional Reconstruction of the Nucleolus Using Meta-Confocal Microscopy in Cells Expressing the Coronavirus Nucleoprotein


title: Microbiologic Diagnostic Workup of Acute Respiratory Failure with Pulmonary Infiltrates after Allogeneic Hematopoietic Stem Cell Transplantation: Findings in the Era of Molecular- and Biomarker-Based Assays

text: Abstract Allogeneic hematopoietic stem cell transplantation (HSCT) recipients frequently develop acute respiratory failure (ARF) with pulmonary infiltrates. Molecular- and biomarker-based assays enhance pathogen detection, but data on their yield in this population are scarce. This was a retrospective single-center study of 156 consecutive HSCT recipients admitted to the intensive care unit (ICU) between May 2013 and July 2017. Findings from a microbiologic diagnostic workup using currently available methods on bronchoalveolar lavage (BAL) and blood samples from 66 patients (age, 58 years [range, 45 to 64]; HSCT to ICU, 176 days [range, 85 to 407]) with ARF and pulmonary infiltrates were analyzed. In 47 patients (71%) a causative pathogen was identified (fungal, n = 28; viral, n = 26; bacterial, n = 18). Polymicrobial findings involving several pathogen groups occurred in 20 patients (30%). Culture (12/16, 75%), galactomannan (13/15, 87%), and Aspergillus-PCR (8/9, 89%) from BAL but not serum galactomannan (6/14, 43%) helped to diagnose invasive aspergillosis (n = 16, 24%). Aspergillus-PCR detected azole resistance in 2 cases. Mucorales was found in 7 patients (11%; BAL culture, n = 6; Mucorales-PCR, n = 1). Patients with identified pathogens had higher Simplified Acute Physiology Score II scores (P = .049) and inferior ICU survival (6% versus 37%, P < .01), which largely related to the presence of an invasive fungal infection. Eight patients (12%) had 1 or more viruses with uncertain lung pathogenicity as the sole microbiologic finding. A diagnostic microbiologic workup incorporating molecular- and biomarker-based assays identified pathogens in most HSCT recipients with ARF and pulmonary infiltrates admitted to the ICU. Implications of polymicrobial infection and pathogen patterns in these patients warrant further investigation.


title: Threat of COVID-19 and emotional state during quarantine: Positive and negative affect as mediators in a cross-sectional study of the Spanish population

text: AIMS: The objective of this study was therefore to analyze the effect of exceptionally stressful situations, such as the current health risk, on the cognitive and emotive state of the individual, that is, perceived threat and emotional state on affect and mood. METHOD: This was a cross-sectional study with snowball sampling. The sample came to 1014 Spanish adults (67.2% women and 32.8% men). The Perception of Threat from COVID-19 questionnaire, the Affective Balance Scale and the Mood Evaluation Scale were used. RESULTS: The results showed that the perception of threat from COVID-19 was related positively to negative affect and emotional signs, that is, sadness-depression, anxiety and anger-hostility. There was a direct positive effect of perceived threat from COVID-19 on sadness-depression, anxiety and anger-hostility moods, while anxiety and anger-hostility had a direct positive effect on perception of threat from the virus. Thus, there was a circular relationship, in which perceived threat influenced the presence of negative mood, and negative mood, in turn, linked to emotions of irritation and agitation from a present situation, promoted the feeling of threat. CONCLUSIONS: A negative affective balance increases both one’s perception of threat from COVID-19 and negative mood. Thus, knowing the emotional and cognitive effects on the population would enable measures to be put into service to facilitate their effective coping.


title: 29 Antiviral drugs

text: Publisher Summary This chapter discusses the adverse effects of antiviral drugs used against cytomegalovirus, herpesviruses, hepatitis viruses, against HIV, and against influenza viruses. The cidofovir, drug active against cytomegalovirus, has been associated with bronchiolitis obliterans. Aciclovir and valaciclovir has been reported with renal insufficiency. Adefovir , a drug active against hepatitis viruses, is associated with the fall in creatinine clearance in patients with lamivudine-resistant HBe antigen (HBeAg)negative disease. Drugs active against HIV are comprehensively reviewed as in combination, nucleoside analogue reverse transcriptase inhibitors, nucleoside analogue reverse transcriptase inhibitors, and protease inhibitors. In a randomized controlled trial of indinavir, saquinavir and lopinavir in combination with low-dose ritonavir in 656 patients, median total cholesterol increased by 0.5 mmol/l in the patients with the highest minimum drug plasma concentrations. In patients with AIDS-associated AIDS dementia complex taking optimal stable background antiretroviral therapy including either abacavir or placebo, there was significantly more nausea in those who took abacavir.


title: Detection of virus particles resembling circovirus and porcine circovirus 2a (PCV2a) sequences in feces of dogs

text: Abstract During routine electron microscopy of fecal samples from diarrheic dogs dated from 2000 virus particles resembling circovirus in shape and size were detected in two samples (V2177/00; V3374/00). Polymerase chain reaction (PCR) using primers specific for porcine circovirus type 2 (PCV2) amplified DNA recovered from both samples. Sequencing of PCR amplificates (V2177/00) obtained with PCV2-specific primer pairs revealed a genome size of 1768bp. The nucleotide sequence was highly similar (98% nucleotide identity) to the PCV2a reference sequence.


title: Controlling epidemic spread by social distancing: Do it well or not at all

text: BACKGROUND: Existing epidemiological models have largely tended to neglect the impact of individual behaviour on the dynamics of diseases. However, awareness of the presence of illness can cause people to change their behaviour by, for example, staying at home and avoiding social contacts. Such changes can be used to control epidemics but they exact an economic cost. Our aim is to study the costs and benefits of using individual-based social distancing undertaken by healthy individuals as a form of control. METHODS: Our model is a standard SIR model superimposed on a spatial network, without and with addition of small-world interactions. Disease spread is controlled by allowing susceptible individuals to temporarily reduce their social contacts in response to the presence of infection within their local neighbourhood. We ascribe an economic cost to the loss of social contacts, and weigh this against the economic benefit gained by reducing the impact of the epidemic. We study the sensitivity of the results to two key parameters, the individuals’ attitude to risk and the size of the awareness neighbourhood. RESULTS: Depending on the characteristics of the epidemic and on the relative economic importance of making contacts versus avoiding infection, the optimal control is one of two extremes: either to adopt a highly cautious control, thereby suppressing the epidemic quickly by drastically reducing contacts as soon as disease is detected; or else to forego control and allow the epidemic to run its course. The worst outcome arises when control is attempted, but not cautiously enough to cause the epidemic to be suppressed. The next main result comes from comparing the size of the neighbourhood of which individuals are aware to that of the neighbourhood within which transmission can occur. The control works best when these sizes match and is particularly ineffective when the awareness neighbourhood is smaller than the infection neighbourhood. The results are robust with respect to inclusion of long-range, small-world links which destroy the spatial structure, regardless of whether individuals can or cannot control them. However, addition of many non-local links eventually makes control ineffective. CONCLUSIONS: These results have implications for the design of control strategies using social distancing: a control that is too weak or based upon inaccurate knowledge, may give a worse outcome than doing nothing.


title: Public health implications of using various case definitions in The Netherlands during the worldwide SARS outbreak

text: This study analysed the consequences of deviation from the WHO case definition for the assessment of patients with suspected severe acute respiratory syndrome (SARS) in The Netherlands during 2003. Between 17 March and 7 July 2003, as a result of dilemmas in balancing sensitivity and specificity, five different case definitions were used. The patients referred for SARS assessment were analysed from a public health perspective. None of the patients referred had SARS, based on serological and virological criteria. Nevertheless, all 72 patients required thorough assessment and, depending on the results of the assessment, institution of appropriate prevention and control measures. Changing case definitions caused confusion in classifying cases. A centralised assessment of the reported cases by a team with clinical and public health expertise (epidemiological and geographical risk assessment) is a practical solution for addressing differences in applying case definitions. The burden of managing non-cases is an important issue when allocating public health resources, and should be taken into account during the preparation phase, rather than during an outbreak. This applies not only to SARS, but also to other public health threats, such as pandemic influenza or a bioterrorist episode.


title: TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells

text: The cellular protease TMPRSS2 cleaves and activates the influenza virus hemagglutinin (HA) and TMPRSS2 expression is essential for viral spread and pathogenesis in mice. Moreover, severe acute respiratory syndrome coronavirus (SARS-CoV) and other respiratory viruses are activated by TMPRSS2. However, previous studies on viral activation by TMPRSS2 focused on a 492 amino acids comprising form of the protein (isoform 2) while other TMPRSS2 isoforms, generated upon alternative splicing of the tmprss2 mRNA, have not been characterized. Here, we show that the mRNA encoding a TMPRSS2 isoform with an extended N-terminal cytoplasmic domain (isoform 1) is expressed in lung-derived cell lines and tissues. Moreover, we demonstrate that TMPRSS2 isoform 1 colocalizes with HA and cleaves and activates HA. Finally, we show that isoform 1 activates the SARS-CoV spike protein for cathepsin L-independent entry into target cells. Our results indicate that TMPRSS2 isoform 1 is expressed in viral target cells and might contribute to viral activation in the host.


title: Epidemic as a natural process

text: Abstract Mathematical epidemiology is a well-recognized discipline to model infectious diseases. It also provides guidance for public health officials to limit outbreaks. Nevertheless, epidemics take societies by surprise every now and then, for example, when the Ebola virus epidemic raged seemingly unrestrained in Western Africa. We provide insight to this capricious character of nature by describing the epidemic as a natural process, i.e., a phenomenon governed by thermodynamics. Our account, based on statistical mechanics of open systems, clarifies that it is impossible to predict accurately epidemic courses because everything depends on everything else. Nonetheless, the thermodynamic theory yields a comprehensive and analytical view of the epidemic. The tenet subsumes various processes in a scale-free manner from the molecular to the societal levels. The holistic view accentuates overarching procedures in arresting and eradicating epidemics.


title: A review of producing renewable aviation fuels from the gasification of biomass and residual wastes

text: This article reviews the production of renewable aviation fuels from biomass and residual wastes using gasification followed by syngas conditioning and Fischer-Tropsch catalytic synthesis. The challenges involved with gasifying wastes are discussed along with a summary conventional and emerging gasification technologies. The techniques for conditioning syngas including removal of particulate matter, tars, sulphur, carbon dioxide and compounds of nitrogen, chlorine and alkali metals are reported. Recent developments in Fischer-Tropsch synthesis, such as new catalyst formulations are described alongside reactor technologies for producing renewable aviation fuels. The energy efficiency and capital cost of converting biomass and residual wastes to aviation fuels are major barriers to widespread adoption. Therefore, further development of advanced technologies will be critical for the aviation industry to achieve their stated greenhouse gas reduction targets by 2050.


A journey through and beyond a “perfect storm”: the COVID-19 pandemic


title: Physico‐Chemical Properties of Mouse Hepatitis Virus (MHV‐2) Grown on DBT Cell Culture

text: Some properties of a strain of mouse hepatitis virus, MHV‐2, grown on DBT cells were determined using a plaque assay on the cells. Viral growth was not inhibited by the presence of actinomycin D or 5‐iodo‐2‐deoxyuridine. MHV‐2 was completely inactivated by ether, chloroform, sodium deoxycholate or beta‐propiolactone, but showed a moderate resistance to trypsin. Heating at 56 C for 30 min did not completely abolish the virus infectivity. The virus was stable after heating at 50 C for 15 min in 1m‐MgCl(2) or 1m‐MgSO(4) as well as at 37 C for 60 min at pH 3.0 to 9.0. Infectivity was decreased to 1/100 and 1/400 after storing at 4 C for 30 days and 37 C for 24 hr, respectively. The virus passed through a 200‐nm but not a 50‐nm Sartorius membrane filter. The buoyant density of MHV‐2 was 1.183 g/cm(3) in sucrose gradient, and the fraction contained coronavirus‐like particles measuring 70 to 130 nm in diameter. Survival rate was 10% after exposure to ultraviolet at 150 ergs/mm(2). Freezing and thawing or sonication at 20 kc for 3 min did not affect the virus titer. No hemagglutinin was demonstrable with red blood cells of the chicken, Japanese quail, mouse, rat, hamster, guinea pig, sheep, bovine or human.


title: Middle East Respiratory Syndrome Coronavirus Antibody Reactors Among Camels in Dubai, United Arab Emirates, in 2005

text: We tested, using a low starting dilution, sequential serum samples from dromedary camels, sheep and horses collected in Dubai from February/April to October of 2005 and from dromedary camels for export/import testing between Canada and USA in 2000–2001. Using a standard Middle East respiratory syndrome coronavirus (MERS-CoV) neutralization test, serial sera from three sheep and three horses were all negative while sera from 9 of 11 dromedary camels from Dubai were positive for antibodies supported by similar results in a MERS-CoV recombinant partial spike protein antibody ELISA. The two negative Dubai camels were both dromedary calves and remained negative over the 5 months studied. The six dromedary samples from USA and Canada were negative in both tests. These results support the recent findings that infection with MERS-CoV or a closely related virus is not a new occurrence in camels in the Middle East. Therefore, interactions of MERS-CoV at the human–animal interface may have been ongoing for several, perhaps many, years and by inference, a widespread pandemic may be less likely unless significant evolution of the virus allow accelerated infection and spread potential in the human population.


title: The impact of within-herd genetic variation upon inferred transmission trees for foot-and-mouth disease virus

text: Abstract Full-genome sequences have been used to monitor the fine-scale dynamics of epidemics caused by RNA viruses. However, the ability of this approach to confidently reconstruct transmission trees is limited by the knowledge of the genetic diversity of viruses that exist within different epidemiological units. In order to address this question, this study investigated the variability of 45 foot-and-mouth disease virus (FMDV) genome sequences (from 33 animals) that were collected during 2007 from eight premises (10 different herds) in the United Kingdom. Bayesian and statistical parsimony analysis demonstrated that these sequences exhibited clustering which was consistent with a transmission scenario describing herd-to-herd spread of the virus. As an alternative to analysing all of the available samples in future epidemics, the impact of randomly selecting one sequence from each of these herds was used to assess cost-effective methods that might be used to infer transmission trees during FMD outbreaks. Using these approaches, 85% and 91% of the resulting topologies were either identical or differed by only one edge from a reference tree comprising all of the sequences generated within the outbreak. The sequence distances that accrued during sequential transmission events between epidemiological units was estimated to be 4.6 nucleotides, although the genetic variability between viruses recovered from chronic carrier animals was higher than between viruses from animals with acute-stage infection: an observation which poses challenges for the use of simple approaches to infer transmission trees. This study helps to develop strategies for sampling during FMD outbreaks, and provides data that will guide the development of further models to support control policies in the event of virus incursions into FMD free countries.


title: A Novel Bacterium-Like Particle-Based Vaccine Displaying the SUDV Glycoprotein Induces Potent Humoral and Cellular Immune Responses in Mice

text: Sudan virus (SUDV) causes severe lethal hemorrhagic fever in humans and nonhuman primates. The most effective and economical way to protect against Sudan ebolavirus disease is prophylactic vaccination. However, there are no licensed vaccines to prevent SUDV infections. In this study, a bacterium-like particle (BLP)-based vaccine displaying the extracellular domain of the SUDV glycoprotein (eGP) was developed based on a gram-positive enhancer matrix-protein anchor (GEM-PA) surface display system. Expression of the recombinant GEM-displayed eGP (eGP-PA-GEM) was verified by Western blotting and immunofluorescence assays. The SUDV BLPs (SBLPs), which were mixed with Montanide ISA 201VG plus Poly (I:C) combined adjuvant, could induce high SUDV GP-specific IgG titers of up to 1:40,960 and robust virus-neutralizing antibody titers reached 1:460. The SBLP also elicited T-helper 1 (Th1) and T-helper 2 (Th2) cell-mediated immunity. These data indicate that the SBLP subunit vaccine has the potential to be developed into a promising candidate vaccine against SUDV infections.


title: Detection of a novel circovirus PCV3 in pigs with cardiac and multi-systemic inflammation

text: BACKGROUND: Porcine circovirus 2 causes different clinical syndromes resulting in a significant economic loss in the pork industry. Three pigs with unexplained cardiac and multi-organ inflammation that tested negative for PCV2 and other known porcine pathogens were further analyzed. METHODS: Histology was used to identify microscopic lesions in multiple tissues. Metagenomics was used to detect viral sequences in tissue homogenates. In situ hybridization was used to detect viral RNA expression in cardiac tissue. RESULTS: In all three cases we characterized the genome of a new circovirus we called PCV3 with a replicase and capsid proteins showing 55 and 35 % identities to the genetically-closest proteins from a bat-feces associated circovirus and were even more distant to those of porcine circovirus 1 and 2. Common microscopic lesions included non-suppurative myocarditis and/or cardiac arteriolitis. Viral mRNA was detected intralesionally in cardiac cells. Deep sequencing in tissues also revealed the presence of porcine astrovirus 4 in all three animals as well as rotavirus A, porcine cytomegalovirus and porcine hemagglutinating encephalomyelitis virus in individual cases. CONCLUSION: The pathogenicity and molecular epidemiology of this new circovirus, alone or in the context of co-infections, warrants further investigations.


title: Modulation of signaling pathways by RNA virus capsid proteins

text: Abstract Capsid proteins are structural components of virus particles. They are nucleic acid-binding proteins whose main recognized function is to package viral genomes into protective structures called nucleocapsids. Research over the last 10 years indicates that in addition to their role as genome guardians, viral capsid proteins modulate host cell signaling networks. Disruption or alteration of intracellular signaling pathways by viral capsids may benefit replication of the virus by affecting innate immunity and in some cases, may underlie disease progression. In this review, we describe how the capsid proteins from medically relevant RNA viruses interact with host cell signaling pathways.


title: Molecular phylogeny of coronaviruses including human SARS-CoV

text: Phylogenetic tree of coronaviruses (CoVs) including the human SARS-associated virus is reconstructed from complete genomes by using our newly developed K-string composition approach. The relation of the human SARS-CoV to other coronaviruses, i.e. the rooting of the tree is suggested by choosing an appropriate outgroup. SARS-CoV makes a separate group closer but still distant from G2 (CoVs in mammalian host). The relation between different isolates of the human SARS virus is inferred by first constructing an ultrametric distance matrix from counting sequence variations in the genomes. The resulting tree is consistent with clinic relations between the SARS-CoV isolates. In addition to a larger variety of coronavirus genomes these results provide phylogenetic knowledge based on independent novel methodology as compared to recent phylogenetic studies on SARS-CoV.


title: The association of being in quarantine and related COVID-19 recommended and non-recommended behaviors with psychological distress in Chinese population

text: • The World Health Organization (WHO) guidelines needs to be empirically examined. • The study sought to look at the recommended and non-recommended behaviors. • The results of the study show that psychological distress was negatively associated with recommended behaviors and positively with non-recommended behaviors.


title: Development of nsP2 protease based cell free high throughput screening assay for evaluation of inhibitors against emerging Chikungunya virus

text: Chikungunya virus has emerged as one of the most important global arboviral threats over the last decade. Inspite of large scale morbidity, with long lasting polyarthralgia, so far no licensed vaccine or antiviral is available. CHIKV nsP2 protease is crucial for processing of viral nonstructural polypeptide precursor to release enzymes required for viral replication, thus making it a promising drug target. In this study, high cell density cultivation (HCDC) of Escherichia coli in batch process was carried out to produce rCHIKV nsP2pro in a cost-effective manner. The purified nsP2pro and fluorogenic peptide substrate have been adapted for fluorescence resonance energy transfer (FRET) based high throughput screening (HTS) assay with Z’ value and CV of 0.67 ± 0.054 and <10% respectively. We used this cell free HTS system to screen panel of metal ions and its conjugate which revealed zinc acetate as a potential candidate, which was further found to inhibit CHIKV in Vero cells. Scale-up process has not been previously reported for any of the arboviral nonstructural enzymes. The successful scale-up method for viral protease together with a HTS assay could lead to the development of industrial level large-scale screening platform for identification of protease inhibitors against emerging and re-emerging viruses.


title: Origin and characteristics of the recombinant novel avian infectious bronchitis coronavirus isolate ck/CH/LJL/111054

text: Abstract Recombination among infectious bronchitis viruses (IBVs), coupled with point mutations, insertions, and deletions that occur in the genome, is thought to contribute to the emergence of new IBV variants. In this study an IBV, ck/CH/LJL/111054, was isolated from a H120-vaccinated chicken, which presented with a suspected IBV infection. Phylogenetic analysis of the S1 subunit sequence confirmed that strain ck/CH/LJL/111054 is of the Connecticut-type; however, further extensive full-length genomic analysis identified the occurrence of recombination events. Therefore, strain ck/CH/LJL/111054 may have originated from recombination events between Conn- and Mass-like strains at three recombination breakpoints: two located within the nsp3 gene sequence and one in the nsp12 gene sequence. Further, the uptake of the 5′ untranslated regions, nsp2, parts of nsp3, nsp4–11, and parts of nsp 12 from Mass-like virus by ck/CH/LJL/111054 might have resulted in changes in viral replication efficiency rather than antigenic changes, via cross-neutralization analysis with the H120 strain. Recombination events coupled with the accumulation of mutations in the ck/CH/LJL/111054 genome may account for its increased virulence in specific-pathogen free chickens.


title: Epidemiological investigation of feline infectious peritonitis in cats living in Harbin, Northeast China from 2017 to 2019 using a combination of an EvaGreen-based real-time RT-PCR and serum chemistry assays

text: Abstract Feline infectious peritonitis (FIP) is caused by the FIP virus (FIPV), a highly virulent mutant form of feline coronavirus (FCoV). This disease is one of the most important infectious diseases in cats, and it is associated with high mortality, particularly among younger cats. In this study, we isolated a wild-type FIPV HRB-17 epidemic strain from the blood sample of household pet cat exhibiting the characteristic wet-form FIP symptoms, which has been confirmed further by animal infection. Further, we developed an EvaGreen-based real-time RT-PCR assay for the accurate detection of FCoV based on the amplification of the highly conserved FIPV N gene. Then, using a combination of the real-time RT-PCR approach and a serum chemistry assay, we performed an epidemiological survey of FIPV infection in cats living in Harbin City, Northeast China. The results indicated that the EvaGreen-based real-time RT-PCR assay can be used for screening FCoV infection in the affected cats at an analytical detection limit of 8.2 × 101 viral genome copies/μL, but could not effectively distinguish FIPVs from FECVs. Additionally, the results of the epidemiological survey investigating feline blood samples (n = 1523) collected between July 2017 to July 2019 revealed an FIPV prevalence of approximately 12% (189/1523). Maybe, the prevalence would be less than 12% due to the real-time RT-PCR assay could not accurately differentiate FIPV and FECV. Nevertheless, it still highlighted the severity of the FIP epidemic in cats and reiterated the urgent need to develop effective anti-FIP therapeutic agents and anti-FIPV vaccines. As pet cats are household animals, risk communication and continuous region-extended surveillance cat programs are recommended.


title: West Nile Virus: Epidemiology, Clinical Presentation, Diagnosis, and Prevention

text: West Nile virus was recognized in the United States for the first time in 1999, when it caused an epidemic of encephalitis and meningitis in New York City, NY. Since then, the disease has been steadily moving westward, and human cases were recognized in 39 states and the District of Columbia in 2002. The infection is caused by a flavivirus that is transmitted from birds to humans through the bite of culicine mosquitoes. Most infections are mild, with symptoms primarily being fever, headache, and myalgias. People older than 50 years are at highest risk of severe disease, which may include encephalomyelitis. In 2002, 5 new modes of transmission were recognized: blood product transfusion, organ transplantation, breast-feeding, transplacental transmission, and occupational exposure in laboratory workers. The transmission season was long, with cases occurring into December in some parts of the United States. Currently, there is no specific drug treatment or vaccine against the infection, and avoiding mosquito bites is the best way to protect against the disease.


A dearth of specifications regarding primary diagnostic monitors (PDMs) for nuclear medicine leaves us with little guidance during the COVID‐19 pandemic


Schwangerschaft, Geburt und Wochenbett mit SARS-CoV-2 und COVID-19


title: COVID‐19: Moving beyond the pandemic

text: Severe acute respiratory syndrome Coronavirus 2 (SARS‐CoV‐2) is the cause of COVID‐19. As of April 11th, 2020, there were almost 2 million cases of COVID‐19 internationally and over 100,000 deaths (John Hopkins University, 2020). There has been significant effort to increase hospital and healthcare capacity to reduce the number of fatalities associated with this global pandemic (Choi & Logsdon 2020).


title: COVID-19 and HELLP: Overlapping Clinical Pictures in Two Gravid Patients

text: Background The impact of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) on pregnancies is currently under investigation. There is a significant overlap between the clinical findings in novel coronavirus disease 2019 (COVID-19) and hemolysis, elevated liver enzymes, and low platelets syndrome (HELLP). Cases Patients presented at 22 and 29 weeks of gestation with suspected COVID-19 pneumonia. While the patient at 22 weeks of gestation subsequently had an intrauterine fetal demise, the patient at 29 weeks of gestation delivered via an emergency cesarean delivery for nonreassuring fetal status. Both patients also developed transaminitis, thrombocytopenia, and disseminated intravascular coagulation with a proof of hemolysis on peripheral smear. Conclusion Clinicians are encouraged to consider both of these diagnoses when caring for pregnant women during the COVID-19 pandemic to assure that both maternal and fetal concerns are addressed and treated appropriately.


title: Arboviruses: A global public health threat

text: Abstract A conference on «ARBOVIRUSES, A GLOBAL PUBLIC HEALTH THREAT» was organized on June 20–22, 2018 at the Merieux Foundation Conference Center in Veyrier du Lac, France, to review and raise awareness to the global public health threat of epidemic arboviruses, and to advance the discussion on the control and prevention of arboviral diseases. The presentations by scientists and public health officials from Asia, the Americas, Europe and Africa strengthened the notion that arboviral diseases of both humans and domestic animals are progressively becoming dominant public health problems in the world. The repeated occurrence of recent deadly epidemics strongly reinforces the call for action against these viral diseases, and the need for developing effective vaccines, drugs, vector control tools and strong prevention programs.


title: Managing healthcare delivery system to fight the COVID-19 epidemic: experience in Japan

text: Amid the global pandemic of a novel Coronavirus Disease 2019 (COVID-19), healthcare delivery system is being stretched. In Japan, rapid spread of the epidemic brings hospitals to the brink of exhaustion. This commentary aims to briefly review related policies of Japan in managing healthcare delivery system. Among the relevant actions, strengthening the hospitalized care is emphasized to save lives. Despite of limitations, the policies show a success in preventing a collapse of healthcare delivery system and skyrocketing mortality from happening so far. On the other hand, huge concerns remain if the infections continue to rapidly increase. The experience in Japan indicates the urgency of planning of healthcare delivery system, mobilizing all relevant social sectors by consensus, and guiding people with calm manner based on the best shared knowledge and evidences.


Online clinical training in the COVID‐19 pandemic


title: Review of Chest CT Manifestations of COVID-19 Infection

text: Coronavirus disease-19 (COVID-19) is a viral pandemic that started in China and has rapidly expanded worldwide. Typical clinical manifestations include fever, cough and dyspnea after an incubation period of 2-14 days. The diagnosis is based on RT-PCR test through a nasopharyngeal swab. Because of the pulmonary tropism of the virus, pneumonia is often encountered in symptomatic patients. Here, we review the pertinent clinical findings and the current published data describing chest CT findings in COVID-19 pneumonia, the diagnostic performance of CT for diagnosis, including differential diagnosis, as well the evolving role of imaging in this disease.


Anxiety persists after recovery from acquired COVID-19 in anaesthesiologists


title: Global aspects of viral glycosylation

text: Enveloped viruses encompass some of the most common human pathogens causing infections of different severity, ranging from no or very few symptoms to lethal disease as seen with the viral hemorrhagic fevers. All enveloped viruses possess an envelope membrane derived from the host cell, modified with often heavily glycosylated virally encoded glycoproteins important for infectivity, viral particle formation and immune evasion. While N-linked glycosylation of viral envelope proteins is well characterized with respect to location, structure and site occupancy, information on mucin-type O-glycosylation of these proteins is less comprehensive. Studies on viral glycosylation are often limited to analysis of recombinant proteins that in most cases are produced in cell lines with a glycosylation capacity different from the capacity of the host cells. The glycosylation pattern of the produced recombinant glycoproteins might therefore be different from the pattern on native viral proteins. In this review, we provide a historical perspective on analysis of viral glycosylation, and summarize known roles of glycans in the biology of enveloped human viruses. In addition, we describe how to overcome the analytical limitations by using a global approach based on mass spectrometry to identify viral O-glycosylation in virus-infected cell lysates using the complex enveloped virus herpes simplex virus type 1 as a model. We underscore that glycans often pay important contributions to overall protein structure, function and immune recognition, and that glycans represent a crucial determinant for vaccine design. High throughput analysis of glycosylation on relevant glycoprotein formulations, as well as data compilation and sharing is therefore important to identify consensus glycosylation patterns for translational applications.


title: Could an endo-lysosomal ion channel be the Achilles heel of SARS-CoV2?

text: Abstract The ongoing SARS-CoV2 outbreak has developed into a global pandemic. Despite previous outbreaks of SARS-CoV and the related MERS-CoV in recent years, neither a vaccine nor any other medication for an effective treatment are currently available. Endo-lysosomal two-pore cation channels have now emerged as potential novel targets for SARS-CoV treatment.


title: Porcine epidemic diarrhea virus (PEDV) introduction into a naive Dutch pig population in 2014

text: Abstract Porcine epidemic diarrhea virus (PEDV) is the highly contagious, causative agent of an economically important acute enteric disease in pigs of all ages. The disease is characterized by diarrhea and dehydration causing mortality and growth retardation. In the last few decades, only classical PEDV was reported sporadically in Europe, but in 2014 outbreaks of PEDV were described in Germany. Phylogenetic analysis showed a very high nucleotide similarity with a variant of PEDV that was isolated in the US in January 2014. The epidemiological situation of PEDV infections in the Netherlands in 2014 was unknown and a seroprevalence study in swine was performed. In total, 838 blood samples from sows from 267 farms and 101 samples from wild boars were collected from May till November 2014 and tested for antibodies against PEDV by ELISA. The apparent herd prevalence of 0.75% suggests that PEDV was not circulating on a large scale in the Netherlands at this time. However, in November 2014 a clinical outbreak of PEDV was diagnosed in a fattener farm by PCR testing. This was the first confirmed PEDV outbreak since the early nineties. Sequence analyses showed that the viruses isolated in 2014 and 2015 in the Netherlands cluster with recently found European G1b strains. This suggests a one event introduction of PEDV G1b strains in Europe in 2014, which made the Netherlands and other European countries endemic for this type of strains since then.


title: Role of tourism price in attracting international tourists: The case of Japanese inbound tourism from South Korea

text: Abstract Tourism price has been extensively used to predict tourism demand. However, there is no agreement on the proper indicators of its components. Use of different price indicators may be the reason for researchers’ apparently inconsistent results. The purpose of this study was to identify proper price indicators for the demand model of Japanese inbound tourism from South Korea. After comparing six models, each with different price indicators, the model with relative price and exchange rate but without transport cost was identified as the best model in which relative price, exchange rate, and per capita income were found to be significant.


title: Estimating clinical severity of COVID-19 from the transmission dynamics in Wuhan, China

text: As of 29 February 2020 there were 79,394 confirmed cases and 2,838 deaths from COVID-19 in mainland China. Of these, 48,557 cases and 2,169 deaths occurred in the epicenter, Wuhan. A key public health priority during the emergence of a novel pathogen is estimating clinical severity, which requires properly adjusting for the case ascertainment rate and the delay between symptoms onset and death. Using public and published information, we estimate that the overall symptomatic case fatality risk (the probability of dying after developing symptoms) of COVID-19 in Wuhan was 1.4% (0.9–2.1%), which is substantially lower than both the corresponding crude or naïve confirmed case fatality risk (2,169/48,557 = 4.5%) and the approximator(1) of deaths/deaths + recoveries (2,169/2,169 + 17,572 = 11%) as of 29 February 2020. Compared to those aged 30–59 years, those aged below 30 and above 59 years were 0.6 (0.3–1.1) and 5.1 (4.2–6.1) times more likely to die after developing symptoms. The risk of symptomatic infection increased with age (for example, at ~4% per year among adults aged 30–60 years).


title: Recycling manure as cow bedding: Potential benefits and risks for UK dairy farms

text: Abstract Material obtained from physical separation of slurry (recycled manure solids; RMS) has been used as bedding for dairy cows in dry climates in the US since the 1970s. Relatively recently, the technical ability to produce drier material has led to adoption of the practice in Europe under different climatic conditions. This review collates the evidence available on benefits and risks of using RMS bedding on dairy farms, with a European context in mind. There was less evidence than expected for anecdotal claims of improved cow comfort. Among animal health risks, only udder health has received appreciable attention. There are some circumstantial reports of difficulties of maintaining udder health on RMS, but no large scale or long term studies of effects on clinical and subclinical mastitis have been published. Existing reports do not give consistent evidence of inevitable problems, nor is there any information on clinical implications for other diseases. The scientific basis for guidelines on management of RMS bedding is limited. Decisions on optimum treatment and management may present conflicts between controls of different groups of organisms. There is no information on the influence that such ‘recycling’ of manure may have on pathogen virulence. The possibility of influence on genetic material conveying antimicrobial resistance is a concern, but little understood. Should UK or other non-US farmers adopt RMS, they are advised to do so with caution, apply the required strategies for risk mitigation, maintain strict hygiene of bed management and milking practices and closely monitor the effects on herd health.


title: Migratory birds-one major source of environmental antibiotic resistance around Qinghai Lake, China

text: Abstract Migratory birds are potential transmitters of bacterial antibiotic resistance. However, their role in the environmental dissemination of bacterial antibiotic resistance and the extent of their impact on the environment are not yet clear. Qinghai Lake is one of the most important breeding and stopover ground for the migratory birds along the Central Asian Flyway. Here, we investigated the bacterial antibiotic resistance in the environment and among the migratory birds around the lake. The results of culture-based analysis of bacterial antibiotic resistance, quantitative PCR and metagenomic sequencing indicate that migratory birds are one major source of bacterial antibiotic resistance in the environment around Qinghai Lake. Network analysis reveals the co-occurrence patterns of antibiotic resistance genes (ARGs) and bacterial genera. Genetic co-localization analysis suggests high co-selection potential (with incidence of 35.8%) among different ARGs, but limited linkage (with incidence of only 3.7%) between ARGs and biocide/metal resistance genes (BMRGs). The high genetic linkage between ARGs and mobile genetic elements (MGEs) is still largely confined to the bacterial community in migratory birds (accounting for 96.0% of sequencing reads of MGE-linked ARGs), which indicates limited horizontal transfer of ARGs to the environment. Nevertheless, the antibiotic resistance determinants carried by migratory birds and their specific genetic properties (high co-selection and mobility potential of the ARGs) remind us that the role of migratory birds in the environmental dissemination of bacterial antibiotic resistance deserves more attention.


title: 17 Approach to the Patient With Malaria

text: Abstract Nearly 3 billion people live at risk of malaria across most of the tropics, subtropics, and even some temperate zones. Millions visit these areas, and each year thousands appear in hospitals with posttravel acute malaria. That diagnosis should be managed as a medical emergency. Illness may deteriorate rapidly without prompt diagnosis and effective treatment. Among five species of Plasmodium responsible for human malaria, Plasmodium falciparum most often deteriorates rapidly, but all species potentially threaten life. Malignant and benign species of malaria parasites is a dangerous fallacy. Clinical malaria mimics other common tropical infections and the diagnosis requires laboratory confirmation, but malaria-like symptoms in a patient exposed to risk should be presumed to be malaria until proven otherwise. Intravenous or intramuscular artesunate is used for severe malaria of any species in any patient, including all trimesters of pregnancy. Primaquine is administered with vivax or ovale malarias after affirming glucose-6-phosphate dehydrogenase (G6PD)–normal status.


title: The Histochem Cell Biol conspectus: the year 2013 in review

text: Herein, we provide a brief synopsis of all manuscripts published in Histochem Cell Biol in the year 2013. For ease of reference, we have divided the manuscripts into the following categories: Advances in Methodologies; Molecules in Health and Disease; Organelles, Subcellular Structures and Compartments; Golgi Apparatus; Intermediate Filaments and Cytoskeleton; Connective Tissue and Extracellular Matrix; Autophagy; Stem Cells; Musculoskeletal System; Respiratory and Cardiovascular Systems; Gastrointestinal Tract; Central Nervous System; Peripheral Nervous System; Excretory Glands; Kidney and Urinary Bladder; and Male and Female Reproductive Systems. We hope that the readership will find this annual journal synopsis of value and serve as a quick, categorized reference guide for “state-of-the-art” manuscripts in the areas of histochemistry, immunohistochemistry, and cell biology.


Rethinking Respiratory Function Test Lab in the Era of COVID-19 : considerations about the “today” and the “day after”


title: Contributions of Latin American researchers in the understanding of the novel coronavirus outbreak: a literature review

text: This article aimed to give the visibility of Latin American researchers’ contributions to the comprehension of COVID-19; our method was a literature review. Currently, the world is facing a health and socioeconomic crisis caused by the novel coronavirus, SARS-CoV-2, and its disease COVID-19. Therefore, in less than 4 months, researchers have published a significant number of articles related to this novel virus. For instance, a search focused on the Scopus database on 10 April 2020, showed 1,224 documents published by authors with 1,797 affiliations from 80 countries. A total of 25.4%, 24.0% and 12.6% of these national affiliations were from China, Europe and the USA, respectively, making these regions leaders in COVID-19 research. In the case of Latin America, on 10 April 2020, we searched different databases, such as Scopus, PubMed and Web of Science, finding that the contribution of this region was 2.7 ± 0.6% of the total publications found. In other words, we found 153 publications related to COVID-19 with at least one Latin American researcher. We summarized and processed the information from these 153 publications, finding active participation in topics like medical, social and environmental considerations, bioinformatics and epidemiology.


title: Reverse transcription loop-mediated isothermal amplification for rapid and quantitative assay of covert mortality nodavirus in shrimp

text: Abstract A disease known as covert mortality disease has become an increasing problem in the shrimp farming industry in recent years in China and several countries of Southeast Asia, leading to serious losses in production. Litopenaeus vannamei (also known as Pacific white shrimp) is affected by this disease that leads to a range of clinical symptoms including hepatopancreas atrophy and necrosis, soft shell, slow growth, and abdominal muscle whitening and necrosis in the acute stage of disease. A new nodavirus, termed covert mortality nodavirus (CMNV), has been shown to be the etiological agent. In this study, we report a sensitive and specific real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the rapid and quantitative detection of CMNV. The optimal conditions for this newly developed RT-LAMP reaction were found to be 6mM MgCl2 and 1.6mM dNTPs, an incubation temperature of 65°C and a reaction time of 50min. The analytical sensitivity of the RT-LAMP assay was estimated to be 6.3pg total RNA of CMNV-infected shrimp and 27 copies of the target plasmid. The diagnostic sensitivity and specificity of the newly developed assay versus the standard nested reverse transcription PCR (RT-PCR) assay was 96.4% and 94.4%, respectively. The reaction products were detected by visual inspection after staining with an in-tube DNA fluorescent dye, a measure taken to eliminate the risk of contamination. The quantitative RT-LAMP assay for CMNV showed high correlation coefficient (r 2 =0.9953) when the initial templates were above 1000 copies, however the correlation coefficient decreased when the initial templates were lower than 1000 copies. Test of viral load in shrimp indicated that the viral loads varied from 1.5×102 to 6.7×106 copies per mg of cephalothorax tissue. Thus, the CMNV RT-LAMP assay is a sensitive and specific new tool for the field detection and quantification of CMNV in the diagnosis and surveillance of covert mortality disease.


title: Structural and Functional Basis of the Fidelity of Nucleotide Selection by Flavivirus RNA-Dependent RNA Polymerases

text: Viral RNA-dependent RNA polymerases (RdRps) play a central role not only in viral replication, but also in the genetic evolution of viral RNAs. After binding to an RNA template and selecting 5′-triphosphate ribonucleosides, viral RdRps synthesize an RNA copy according to Watson-Crick base-pairing rules. The copy process sometimes deviates from both the base-pairing rules specified by the template and the natural ribose selectivity and, thus, the process is error-prone due to the intrinsic (in)fidelity of viral RdRps. These enzymes share a number of conserved amino-acid sequence strings, called motifs A–G, which can be defined from a structural and functional point-of-view. A co-relation is gradually emerging between mutations in these motifs and viral genome evolution or observed mutation rates. Here, we review our current knowledge on these motifs and their role on the structural and mechanistic basis of the fidelity of nucleotide selection and RNA synthesis by Flavivirus RdRps.


title: A long distance RT-PCR able to amplify the Pestivirus genome

text: Abstract A method to amplify long genomic regions (up to ∼12.3kb) from pestiviruses in one RT-PCR is described. The difficulty in designing conserved Pestivirus primers for the amplification of genomes from highly divergent isolates simply by means of overlapping segments is demonstrated using new bioinformatic tools. An alternative procedure consisting of optimizing the length of the genomic cDNA fragments and their subsequent amplification by polymerase chain reaction (PCR) using a limited set of specific primers is described. The amplification of long DNA fragments from a variety of sources, including genomic, mitochondrial, and viral DNAs as well as cDNA produced by reverse transcription (RT) has been achieved using this methodology, known as long distance PCR. In the case of viruses, it is necessary to obtain viral particles from infected cells prior to RT procedures. This work provides improvements in four steps of long distance RT-PCR (L-RT-PCR): (i) preparation of a viral stock, (ii) preparation of template RNA, (iii) reverse transcription and (iv) amplification of the cDNA by LD-PCR. The usefulness of L-RT-PCR is discussed in the light of current knowledge on pestivirus diversity. The genomic sequence of Singer_Arg reference strain obtained using this method is presented and characterized.


title: Serine 105 and 120 are important phosphorylation sites for porcine reproductive and respiratory syndrome virus N protein function

text: Abstract The nucleocapsid (N) protein is the most abundant protein of porcine reproductive and respiratory syndrome virus (PRRSV). It has been shown to be multiphosphorylated. However, the phosphorylation sites are still unknown. In this study, we used liquid chromatography tandem mass spectrometry (LC–MS/MS) to analyze the phosphorylation sites of N protein expressed in Sf9 cells. The results showed that N protein contains two phosphorylation sites. Since N protein can regulate IL-10, which may facilitate PRRSV replication, we constructed four plasmids (pCA-XH-GD, pCA-A105, pCA-A120 and pCA-A105-120) and transfected them into Pig alveolar macrophages (PAMs,3D4/2). The qPCR results showed that mutations at residues 105 and 120 were associated with down-regulation of the IL-10 mRNA level, potentially decreasing the viral growth ability. Then, we mutated the phosphorylation sites (S105A and S120A) and rescued three mutated viruses, namely, A105, A120 and A105-120. Compared with wild-type virus titers, the titers of the mutated viruses at 48 hpi were significantly decreased. The Nsp(non-structural protein) 9 qPCR results were consistent with the multistep growth kinetics results. The infected PAMs (primary PAMs) results were similar with Marc-145.The findings indicated that the mutations impaired the viral replication ability. The confocal microscopy results suggested that mutations to residues 105 and 120 did not affect N protein distribution. Whether the mutations affected other functions of N protein and what the underlying mechanisms are need further investigation. In conclusion, our results show that residues 105 and 120 are phosphorylation sites and are important for N protein function and for viral replication ability.


title: Early inhibitors of human cytomegalovirus: State-of-art and therapeutic perspectives

text: Abstract Human cytomegalovirus (HCMV) infection is associated with severe morbidity and mortality in immunocompromised individuals, mainly transplant recipients and AIDS patients, and is the most frequent cause of congenital malformations in newborn children. To date, few drugs are licensed for the treatment of HCMV infections, most of which target the viral DNA polymerase and suffer from many drawbacks, including long-term toxicity, low potency, and poor bioavailability. In addition, the emergence of drug-resistant viral strains is becoming an increasing problem for disease management. Finally, none of the current anti-HCMV drugs have been approved for the treatment of congenital infections. For all these reasons, there is still a strong need for new anti-HCMV drugs with novel mechanisms of action. The first events of the virus replication cycle, including attachment, entry, immediate-early gene expression, and immediate-early functions—in particular that of Immediate-Early 2 protein—represent attractive targets for the development of novel antiviral compounds. Such inhibitors would block not only the expression of viral immediate-early proteins, which play a key role in the pathogenesis of HCMV infection, but also the host immunomodulation and the changes to cell physiology induced by the first events of virus infection. This review describes the current knowledge on the initial phases of HCMV replication, their validation as potential novel antiviral targets, and the development of compounds that block such processes.


title: Behind the mask: Determinants of nurse's adherence to facial protective equipment

text: BACKGROUND: As the predominant occupation in the health sector and as the health worker with the most patient interaction, nurses are at high risk for occupational transmission of communicable respiratory illness. The use of facial protective equipment (FPE) is an important strategy to prevent occupational transmission. METHODS: A 2-phased study was conducted to examine nurse's adherence to recommended use of FPE. Phase 1 was a cross-sectional survey of nurses in selected units of 6 acute care hospitals in Toronto, Canada. Phase 2 was a direct observational study of critical care nurses. RESULTS: Of the 1,074 nurses who completed surveys (82% response rate), 44% reported adherence to recommended use of FPE. Multivariable analysis revealed 6 predictors of adherence: unit type, frequency of equipment use, equipment availability, training, organizational support, and communication. Following the survey, 100 observations in 14 intensive care units were conducted that revealed a 44% competence rate with proper use of N95 respirators and knowledge as a significant predictor of competence. CONCLUSION: Whereas increasing knowledge should enhance competence, strategies to improve adherence to recommended use of FPE in a busy and complex health care setting should focus on ready availability of equipment, training and fit testing, organizational support for worker health and safety, and good communication practices.


title: Coronavirus derived expression systems

text: Abstract Both helper dependent expression systems, based on two components, and single genomes constructed by targeted recombination, or by using infectious cDNA clones, have been developed. The sequences that regulate transcription have been characterized mainly using helper dependent expression systems and it will now be possible to validate them using single genomes. The genome of coronaviruses has been engineered by modification of the infectious cDNA leading to an efficient (>20 μg ml−1) and stable (>20 passages) expression of the foreign gene. The possibility of engineering the tissue and species tropism to target expression to different organs and animal species, including humans, increases the potential of coronaviruses as vectors. Thus, coronaviruses are promising virus vectors for vaccine development and, possibly, for gene therapy.


title: COVID-19 and Stroke: Incidental, Triggered or Causative

text: Stroke is a serious neurological comorbidity observed during the ongoing COVID-19 (coronavirus associated disease 2019) pandemic caused by SARS-CoV-2 (severe acute respiratory syndrome, corona virus 2) and includes ischemic stroke, intracerebral haemorrhage and cerebral venous thrombosis. We reviewed factors that could potentially contribute to the occurrence of stroke among patients with COVID-19. There could be an interaction between the conventional risk factors of stroke, infection, systemic inflammatory response and plaque destabilisation. Inflammatory markers, D-dimer elevation and increased cytokine activity have been observed in patients with COVID-19. Other probable contributing factors include cardiac injury leading to embolism, a prothrombotic state and a possibility of direct neuroinvasive potential causing vasculopathy. Data from stroke patients gathered in large multicentric cohorts could help shed more light on the occurrence, behaviour, aetiology, pathophysiology, biomarkers and outcomes of stroke occurring during the ongoing pandemic.


title: Experimental Infection of Piglets with a Korean Strain of Porcine Epidemic Diarrhoea Virus

text: Abstract Thirty colostrum-deprived piglets aged 1 day were inoculated with a Korean strain of porcine epidemic diarrhoea virus (PEDV). The purpose was to elucidate the pathogenicity and viral distribution in PEDV-infected piglets over a period of 60 h, by morphometric analysis, in-situ hybridization and immunohistochemistry. At 24–60 h post-inoculation (hpi), the villous height/crypt depth (VH/CD) ratio of infected pigs was significantly less than that of control pigs. Positive cells typically exhibited a dark black reaction product (in-situ hybridization) or brown reaction product (immunohistochemistry) in the cytoplasm, without background staining. PEDV nucleic acid and antigen were detected in the duodenum, jejunum and ileum of experimentally infected pigs. The results suggested that the Korean strain was virulent and caused severe villous atrophy in the small intestine.


The risk of respiratory tract infections and symptoms in psoriasis patients treated with IL-17-pathway inhibiting biologics: A meta-estimate of pivotal trials relevant to decision-making during the COVID-19 pandemic


title: Aerosol generating procedures and infective risk to healthcare workers: SARS-CoV-2 – the limits of the evidence

text: The transmission behaviour of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still being defined. It is likely that it is transmitted predominantly by droplets and direct contact and it is possible that there is at least opportunistic airborne transmission. In order to protect healthcare staff adequately is necessary that we establish whether aerosol generating procedures (AGPs) increase the risk of transmission of SARS-CoV-2. Where we do not have evidence relating to SARS-CoV-2, guidelines for safely conducting these procedures should consider what risk procedures would have of transmitting related pathogens. Currently there is very little evidence detailing the transmission of SARS-CoV-2 associated with any specific procedures. Regarding aerosol generating procedures and respiratory pathogens in general, there is still a large knowledge gap that will leave clinicians unsure what risk they are putting themselves in when offering these procedures. This review aimed to summarise the evidence (and gaps in evidence) around AGPs and SARS-CoV-2.


title: The burden of severe cases of Influenza disease: the Friuli Venezia Giulia Region experience

text: INTRODUCTION: Influenza is a matter of serious concern for clinicians, in both outpatient and in-hospital settings. Worldwide, the 2017-18 epidemic proved to be the most severe since 2003-04. We report a real-world experience regarding the management of patients with influenza admitted to a large teaching hospital in the Friuli Venezia Giulia region during the 2017-2018 influenza season. We also provide a practical guide for the management of hospitalized influenza patients. METHODS: A retrospective observational analysis was conducted among all influenza patients requiring admission to our center during the 2017-18 season. RESULTS: Overall, 29 patients were admitted to the University Hospital of Udine during the 2017-18 season with a diagnosis of influenza. B virus was responsible for the majority of cases. More than 65.5% of the subjects presented with a complication. We estimated that 41.4% of the patients admitted were affected by a “severe form”. All these cases required admission to the Intensive Care Unit, with 27.6% and 10.3% needing Orotracheal Intubation and Extracorporeal Membrane Oxygenation, respectively. The fatality rate was 24.1%. Notably, only 9 subjects in our cohort had been vaccinated. Based on the experience acquired during the past season, we propose a practical guide to the management of influenza cases in everyday hospital practice. CONCLUSION: The cornerstones of the management of all hospitalized influenza patients are the rapid identification and treatment of severe forms. Timely and strict adherence to contact and respiratory precautions are also fundamental to reducing the risk of intra-hospital outbreaks. Despite improvements in antiviral therapies and supportive measures, influenza-related morbidity and mortality remain high. In our opinion, a universal vaccination program is the only safe and effective method of filling the gap.


title: Rapid detection of viral antibodies based on multifunctional Staphylococcus aureus nanobioprobes

text: Abstract Biosynthesis of nanoparticles inside S. aureus cells has enhanced the sensitivity of immunoassays based on the S. aureus nanoparticles. However, the current methods are limited to antigen detection by conjugating IgG antibodies on S. aureus nanoparticles. In this study, a simple way to conjugate antigens to the S. aureus nanobioparticles was developed by utilizing a cell wall binding domain (CBD) from a bacteriophage lysin PlyV12. Based on this novel design, simple agglutination tests of the IgG antibodies of Ebola virus (EBOV) nucleoprotein (NP) and Middle East Respiratory Virus (MERS) NP in rabbit sera were successfully developed by conjugating the S. aureus nanobioparticles with two fusion proteins EBOV NP- CBD and MERS NP-CBD, respectively. The conjugation was done easily by just mixing the fusion proteins with the S. aureus nanoparticles. The detection time was within 20 min without any special equipment or expertise. As far as we know, this is the first time to realize the detection of viral antibodies based on S. aureus nanoparticles.


title: Indicator organisms for estuarine and marine waters

text: Abstract The use of indicator organisms for estuarine and coastal waters has been reviewed. The natural flora of the environment must be considered in selecting an indicator organism, but, more importantly, recent work which shows a viable but non-recoverable stage of pathogens entering the marine environment demonstrates that the conventional detection of indicator microorganisms is misleading, if not inaccurate. Results suggest that the newly developed epifluorescent/immunofluorescent direct detection of pathogens in the environment may be the most reliable method for determining public health hazards in marine and estuarine waters.


title: Linkages between animal and human health sentinel data

text: INTRODUCTION: In order to identify priorities for building integrated surveillance systems that effectively model and predict human risk of zoonotic diseases, there is a need for improved understanding of the practical options for linking surveillance data of animals and humans. We conducted an analysis of the literature and characterized the linkage between animal and human health data. We discuss the findings in relation to zoonotic surveillance and the linkage of human and animal data. METHODS: The Canary Database, an online bibliographic database of animal-sentinel studies was searched and articles were classified according to four linkage categories. RESULTS: 465 studies were identified and assigned to linkage categories involving: descriptive, analytic, molecular, or no human outcomes of human and animal health. Descriptive linkage was the most common, whereby both animal and human health outcomes were presented, but without quantitative linkage between the two. Rarely, analytic linkage was utilized in which animal data was used to quantitatively predict human risk. The other two categories included molecular linkage, and no human outcomes, which present health outcomes in animals but not humans. DISCUSSION: We found limited use of animal data to quantitatively predict human risk and listed the methods from the literature that performed analytic linkage. The lack of analytic linkage in the literature might not be solely related to technological barriers including access to electronic database, statistical software packages, and Geographical Information System (GIS). Rather, the problem might be from a lack of understanding by researchers of the importance of animal data as a 'sentinel' for human health. Researchers performing zoonotic surveillance should be aware of the value of animal-sentinel approaches for predicting human risk and consider analytic methods for linking animal and human data. Qualitative work needs to be done in order to examine researchers' decisions in linkage strategies between animal and human data.


title: Complete nucleotide sequence of polyprotein gene 1 and genome organization of turkey coronavirus

text: Abstract The complete nucleotide sequence of polyprotein gene 1 and the assembled full-length genome sequence are presented for turkey coronavirus (TCoV) isolates 540 and ATCC. The TCoV polyprotein gene encoded two open reading frames (ORFs), which are translated into two products, pp1a and pp1ab, the latter being produced via −1 frameshift translation. TCoV polyprotein pp1a and pp1ab were predicted to be processed to 15 non-structure proteins (nsp2–nsp16), with nsp1 missing. ClustalW analysis revealed 88.99% identity and 96.99% similarity for pp1ab between TCoV and avian infectious bronchitis virus (IBV) at the amino acid level. The whole genome consists of 27,749nucleotides for 540 and 27,816nucleotides for ATCC, excluding the poly(A) tail. A total of 13 ORFs were predicted for TCoV. Five subgenomic RNAs were detected from ATCC-infected turkey small intestines by Northern blotting. The whole genome sequence had 86.9% identity between TCoV and IBV, supporting that TCoV is a group 3 coronavirus.


title: SARS-CoV-2 isolation and propagation from Turkish COVID-19 patients

text: The novel coronavirus pneumonia, which was named later as coronavirus disease 2019 (COVID-19), is caused by the severe acute respiratory syndrome coronavirus 2, namely SARS-CoV-2. It is a positive-strand RNA virus that is the seventh coronavirus known to infect humans. The COVID-19 outbreak presents enormous challenges for global health behind the pandemic outbreak. The first diagnosed patient in Turkey has been reported by the Republic of Turkey Ministry of Health on March 11, 2020. In May, over 150,000 cases in Turkey, and 5.5 million cases around the world have been declared. Due to the urgent need for a vaccine and antiviral drug, isolation of the virus is crucial. Here, we report 1 of the first isolation and characterization studies of SARS-CoV-2 from nasopharyngeal and oropharyngeal specimens of diagnosed patients in Turkey. This study provides an isolation and replication methodology,and cell culture tropism of the virus that will be available to the research communities.


title: Responses of the Toll-like receptor and melanoma differentiation-associated protein 5 signaling pathways to avian infectious bronchitis virus infection in chicks

text: Avian infectious bronchitis virus (IBV) is a Gammacoronavirus in the family Coronaviridae and causes highly contagious respiratory disease in chickens. Innate immunity plays significant roles in host defense against IBV. Here, we explored the interaction between IBV and the host innate immune system. Severe histopathological lesions were observed in the tracheal mucosa at 3-5 days post inoculation (dpi) and in the kidney at 8 dpi, with heavy viral loads at 1-11 and 1-28 dpi, respectively. The expression of mRNAs encoding Toll-like receptor (TLR) 3 and TLR7 were upregulated at 3-8 dpi, and that of TIR-domain-containing adapter-inducing interferon (IFN) β (TRIF) was upregulated at 21 dpi in the trachea and kidney. Myeloid differentiation primary response protein 88 (MyD88) was upregulated in the trachea during early infection. Tumor necrosis factor receptor-associated factor (TRAF) 3 and TRAF6 were upregulated expression in both tissues. Moreover, melanoma differentiation-associated protein 5 (MDA5), laboratory of genetics and physiology 2 (LGP2), stimulator of IFN genes (STING), and mitochondrial antiviral signaling protein (MAVS), as well as TANK binding kinase 1 (TBK1), inhibitor of kappaB kinase (IKK) ε, IKKα, IKKβ, IFN regulatory factor (IRF) 7, nuclear factor of kappaB (NF-ĸB), IFN-α, IFN-β, various interleukins(ILs), and macrophage inflammatory protein-1β (MIP-1β) were significantly upregulated in the trachea and downregulated in the kidney. These results suggested that the TLR and MDA5 signaling pathways and innate immune cytokine were induced after IBV infection. Additionally, consistent responses to IBV infection were observed during early infection, with differential and complicated responses in the kidney. [Image: see text]


title: Ginkgo leaf extract and dipyridamole injection for chronic cor pulmonale: a PRISMA-compliant meta-analysis of randomized controlled trials

text: Ginkgo leaf extract and dipyridamole injection (GLED), a kind of Chinese herbal medicine preparation, has been considered as a promising supplementary treatment for chronic cor pulmonale (CCP). Although an analysis of the published literature has been performed, the exact effects and safety of GLED have yet to be systematically investigated. Therefore, a wide-ranging systematic search of electronic databases from which to draw conclusions was conducted. All randomized controlled trials concerning the GLED plus conventional treatments for CCP were selected in the present study. Main outcomes were treatment efficacy, blood gas and hemorrheology indexes, and adverse events. Data from 28 trials with 2457 CCP patients were analyzed. The results indicated that, compared with conventional treatments alone, the combination of conventional treatments with GLED obviously improved the markedly effective rate (RR = 1.44, 95% CI = 1.31–1.58, P < 0.00001) and total effective rate (RR = 1.28, 95% CI = 1.18–1.38, P < 0.00001). Moreover, the hemorrheology (PaO(2), P < 0.00001; PaCO(2), P < 0.00001; SaO(2), P < 0.00001; pH value, P = 0.05) and blood gas indexes (PV, WBHSV, WBMSV, WBLSV, hematocrit and FBG, P < 0.01) of CCP patients were also significantly ameliorated after the combined therapy. The frequency of adverse events did not differ significantly between the two groups (P > 0.05). In summary, evidence from the meta-analysis suggested that the combination of conventional treatments and GLED appeared to be effective and relatively safe for CCP. Therefore, GLED mediated therapy could be recommended as an adjuvant treatment for CCP.