title: Identification of probable genomic packaging signal sequence from SARS-CoV genome by bioinformatics analysis.

text: AIM To predict the probable genomic packaging signal of SARS-CoV by bioinformatics analysis. The derived packaging signal may be used to design antisense RNA and RNA interfere (RNAi) drugs treating SARS. METHODS Based on the studies about the genomic packaging signals of MHV and BCoV, especially the information about primary and secondary structures, the putative genomic packaging signal of SARS-CoV were analyzed by using bioinformatic tools. Multi-alignment for the genomic sequences was performed among SARS-CoV, MHV, BCoV, PEDV and HCoV 229E. Secondary structures of RNA sequences were also predicted for the identification of the possible genomic packaging signals. Meanwhile, the N and M proteins of all five viruses were analyzed to study the evolutionary relationship with genomic packaging signals. RESULTS The putative genomic packaging signal of SARS-CoV locates at the 3' end of ORF1b near that of MHV and BCoV, where is the most variable region of this gene. The RNA secondary structure of SARS-CoV genomic packaging signal is very similar to that of MHV and BCoV. The same result was also obtained in studying the genomic packaging signals of PEDV and HCoV 229E. Further more, the genomic sequence multi-alignment indicated that the locations of packaging signals of SARS-CoV, PEDV, and HCoV overlaped each other. It seems that the mutation rate of packaging signal sequences is much higher than the N protein, while only subtle variations for the M protein. CONCLUSIONS The probable genomic packaging signal of SARS-CoV is analogous to that of MHV and BCoV, with the corresponding secondary RNA structure locating at the similar region of ORF1b. The positions where genomic packaging signals exist have suffered rounds of mutations, which may influence the primary structures of the N and M proteins consequently.


title: Prevalence of SARS-CoV antibody in all Hong Kong patient contacts.

text: The near absence of transmission (seroprevalence=0.19%) resulting in asymptomatic infection in this representative high-risk group of close contacts indicates that the prevailing SARS-CoV strains in Hong Kong almost always led to clinically apparent disease.


title: Infection of the basal ganglia by a murine coronavirus.

text: The coronavirus, mouse hepatitis virus strain A59 (MHV-A59), causes mild encephalitis and chronic demyelination. Immunohistochemical techniques showed that MHV-A59-infected C57BL/6 mice contained dense deposits of viral antigen in the subthalamic nucleus and substantia nigra, with fewer signs of infection in other regions of the brain. The animals showed extra- and intracellular vacuolation, neuronal loss, and gliosis in the subthalamic-nigral region. Such localization is unprecedented among known viral encephalitides of humans and other species. This infection by a member of a viral class capable of causing both encephalitis and persistent infection in several species may be related to postencephalitic parkinsonism.


Severe acute respiratory syndrome (SARS) in elders.


Estimated timing of the last common ancestor of the SARS coronavirus.


title: Interdisciplinary problem-based learning as a method to prepare Micronesia for public health emergencies.

text: CONTEXT The University of Hawai'i Pacific Basin Bioterrorism Curriculum Development Project has developed a problem-based learning (PBL) curriculum for teaching health professionals and health professional students about bioterrorism and other public health emergencies. These PBL cases have been incorporated into interdisciplinary training settings in community-based settings, such as in the small island districts of the U.S.-Affiliated Pacific Islands. METHODS Quantitative and qualitative methods have been utilized in the evaluation of the PBL cases, PBL tutorials, and the accomplishment of learning objectives. FINDINGS Evaluation of the PBL tutorials demonstrates that PBL is an educational and training modality appropriate for such settings. Participants found it helpful to learn in interdisciplinary groups. The educational process was modified in accordance with local culture. CONCLUSION PBL is a useful educational modality for settings where healthcare staffing and available resources are limited.


[Dilated cardiomyopathy].


title: Evaluating viral interference between infectious bronchitis virus and Newcastle disease virus vaccine strains using quantitative reverse transcription-polymerase chain reaction.

text: The potential for infectious bronchitis virus (IBV) and Newcastle disease virus (NDV) replication interference was evaluated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Fourteen-day-old broiler chickens were inoculated via eyedrop with live commercial vaccine strains of IBV and NDV alone or in combination to directly evaluate IBV and NDV replication in the trachea at 1, 3, and 5 days after vaccination. Commercial NDV vaccine strains used were B1, VG/GA, and C2. The vaccine strains of IBV tested were Massachusetts (Mass) and Arkansas (Ark). The NDV + Mass vaccines used were commercially manufactured combined products. The NDV + Ark vaccines used were commercial vaccines manufactured as single entity products that were administered by eyedrop to opposite eyes of each chicken. As measured by qRT-PCR, the replication of NDV strains B1, VG/GA, and C2 did not interfere with the growth of IBV Mass and Ark strain vaccines in the combined vaccine treatment groups. Combination vaccinations using B1 and VG/GA did not interfere with IBV immunity based on challenge or serum antibody production. In the C2 + Mass vaccination trial, IBV immunity after challenge was reduced, but it did not seem to be a result of reduced Mass vaccine growth or the ability of the Mass vaccine to induce serum IBV antibody. In contrast, the replication of IBV strains Mass and Ark interfered with the growth of NDV strains B1, VG/GA, and C2 as measured by qRT-PCR. However, interference with NDV replication was not reflected in a reduction in Newcastle disease challenge of immunity findings when combination Mass + NDV products manufactured by vaccine companies were tested. Moreover, NDV immunity was not compromised in two of three trials using single entity vaccines of NDV and Ark IBV vaccines manufactured separately but administered simultaneously. However, in one trial, NDV immunity was decreased where a NDV single entity product (C2) was given with an IBV single entity Ark vaccine. This finding emphasizes the importance of using manufactured combination vaccines whenever possible to avoid potential interference.


title: Clinical Features and Outcome of SARS-CoV-2 Infection in Children: A Systematic Review and Meta-analysis.

text: BACKGROUND AND OBJECTIVE Knowledge about COVID-19 in children is limited due to the paucity of reported data. The pediatric age group comprises only less than 5% of total COVID-19 worldwide, therefore, large studies in this population are unlikely in the immediate future. Hence, we planned to synthesize the current data that will help in a better understanding of COVID-19 in children. METHODS Four different electronic databases (MEDLINE, EMBASE, Web of Science, and CENTRAL) were searched for articles related to COVID-19 in the pediatric population. We included studies reporting disease characteristics and outcomes of COVID-19 in patients aged less than 19 years. We performed a random-effect meta-analysis to provide pooled estimates of various disease characteristics. RESULTS 27 studies (4857 patients) fulfilling the eligibility criteria were included in this systematic review, from a total of 883 records. About half of the patients had each of fever and cough, 11% (6-17%) had fast breathing, and 6-13% had gastrointestinal manifestations. Most of the patients had mild to moderate disease, and only 4% had a severe or critical illness. Leukopenia was the commonest reported laboratory abnormality. CONCLUSION Even among the symptomatic COVID-19 cases, severe manifestations are seen in very few children. Though fever and respiratory symptoms are most common, many children also have gastrointestinal manifestations.


title: Stockpersons' attitudes to the husbandry of dairy cows.

text: The health and welfare of dairy cows are dependent upon the stockpersons who handle, observe and monitor them. The attitudes and behaviour of these people have been little studied, although they are fundamental for the animals' well-being and performance. The results of a series of standardised interviews with 238 dairy stockpersons are discussed. Milking was considered both the most important and the most enjoyed task, and the most disliked task was cleaning the parlour and yards. These attitudes can affect the health of the herd. The stockpersons valued their interaction with animals; 'a special event involving the cows' was frequently mentioned as the component of a 'good' day, and a 'bad event involving the cows' was almost unanimously the factor associated with a 'bad' day.


title: Flavonoids and linderone from Lindera oxyphylla and their bioactivities.

text: A new linderone A, namely 2-cinnamoyl-3-hydroxy-4, 5-dimethoxycyclopenta-2, 4-dienone (5), together with three known flavonoids (1-3) and one linderone (4), were isolated from the bark of Lindera oxyphylla. Extensive spectroscopic analysis including 1D and 2D-NMR spectra determined their sturctures. In addition, the antioxidant activity of all the compounds has been determined using 2, 2-diphenyl-1-picrylhydrazyl radical scavenging (DPPH), ferric reducing antioxidant power (FRAP) and ferrous ion chelating (FIC) methods. Compound 3 showed excellent DPPH scavenging activity with IC50% value of 8.5 ± 0.004% (μg/mL) which is comparable with vitamin C. This compound, also showed an absorbance value of 1.00 ± 0.06% through FRAP test when compared with Butyl Hydroxy Aniline (BHA). However, FIC showed low activity for all the isolated compounds (chelating activity less than 50%) in comparison with ethylene diamine tetra acetic acid (EDTA). Anticancer activity for all compounds has also been measured on A375 human melanoma, HT-29 colon adenocarcinoma, MCF-7 human breast adenocarcinoma cells, WRL-68 normal hepatic cells, A549 non-small cell lung cancer cells and PC-3 prostate adenocarcinoma cell line. Compound 1 showed A549=65.03%, PC-3=30.12%, MCF-7=47.67, compound 2 showed PC-3=90.13%, compound 3 showed MCF-7=79.57 and for compound 5 MCF-7 is 96.33.


title: Inhibition of human coronavirus NL63 infection at early stages of the replication cycle.

text: Human coronavirus NL63 (HCoV-NL63), a recently discovered member of the Coronaviridae family, has spread worldwide and is associated with acute respiratory illness in young children and elderly and immunocompromised persons. Further analysis of HCoV-NL63 pathogenicity seems warranted, in particular because the virus uses the same cellular receptor as severe acute respiratory syndrome-associated coronavirus. As there is currently no HCoV-NL63-specific and effective vaccine or drug therapy available, we evaluated several existing antiviral drugs and new synthetic compounds as inhibitors of HCoV-NL63, targeting multiple stages of the replication cycle. Of the 28 compounds that we tested, 6 potently inhibited HCoV-NL63 at early steps of the replication cycle. Intravenous immunoglobulins, heptad repeat 2 peptide, small interfering RNA1 (siRNA1), siRNA2, beta-D-N(4)-hydroxycytidine, and 6-azauridine showed 50% inhibitory concentrations of 125 microg/ml, 2 microM, 5 nM, 3 nM, 400 nM, and 32 nM, respectively, and low 50% cytotoxicity concentrations (>10 mg/ml, >40 microM, >200 nM, >200 nM, >100 microM, and 80 microM, respectively). These agents may be investigated further for the treatment of coronavirus infections.


title: Murine coronavirus packaging signal confers packaging to nonviral RNA.

text: Studies of defective interfering (DI) RNAs of the murine coronavirus mouse hepatitis virus (MHV) suggest that a 69-nucleotide-long packaging signal is necessary for MHV genomic RNA packaging into MHV particles. In this study we showed that when RNA transcripts that consisted of a non-MHV sequence and the packaging signal were expressed in MHV-infected cells, they were packaged into MHV particles. Those RNA transcripts that lacked the packaging signal or those containing a mutated packaging signal did not package efficiently. Thus, the presence of the packaging signal was sufficient for RNA packaging into MHV particles.


title: The prevalence of a group 2 coronavirus in dogs in Japan.

text: Canine coronavirus (CCoV) has been reported to cause acute diarrhea mainly in young pups. CCoV and feline coronavirus are classified as group 1 coronaviruses. However, it has recently been reported in the United Kingdom that the group 2 coronavirus gene, which is more closely related to the bovine coronavirus (BCoV) and human coronavirus strain OC43, has been detected in respiratory tract tissue samples from dogs with respiratory disease. In this study, we examined the prevalence of antibodies to group 2 coronaviruses in domestic dogs and cats in Japan by a neutralization test using BCoV. All 104 feline serum samples were negative (<1:5) for anti-BCoV antibodies. In contrast, of the 898 canine serum samples, 160 (17.8%) were positive for anti-BCoV antibodies, and the antibody titers ranged from 1:5 to more than 1:640, with 1:160 being the most frequent. No correlation was found between the titers of the anti-BCoV and anti-CCoV antibodies in the 198 serum samples of dogs with a known history of CCoV vaccination. We amplified, by RT-PCR, group 2 coronavirus-specific hemagglutination/esterase genes in the oral swabs of a total of 10 young pups presenting with or having recovered from respiratory signs, or having anti-BCoV antibodies, with the result that 2 pups were positive for the hemagglutination/esterase genes. These results strongly suggest that an unknown group 2 coronavirus as well as the known enteritis-causing CCoV (group 1 coronavirus) is prevalent among domestic dogs in Japan.


title: Safety and long-term effects of renal denervation: Rationale and design of the Dutch registry.

text: BACKGROUND Percutaneous renal denervation (RDN) has recently been introduced as a treatment for therapy-resistant hypertension. Also, it has been suggested that RDN may be beneficial for other conditions characterised by increased sympathetic nerve activity. There are still many uncertainties with regard to efficacy, safety, predictors for success and long-term effects. To answer these important questions, we initiated a Dutch RDN registry aiming to collect data from all RDN procedures performed in the Netherlands. METHODS The Dutch RDN registry is an ongoing investigator-initiated, prospective, multicentre cohort study. Twenty-six Dutch hospitals agreed to participate in this registry. All patients who undergo RDN, regardless of the clinical indication or device that is used, will be included. Data are currently being collected on eligibility and screening, treatment and follow-up. RESULTS Procedures have been performed since August 2010. At present, data from 306 patients have been entered into the database. The main indication for RDN was hypertension (n = 302, 99%). Patients had a mean office blood pressure of 177/100 (±29/16) mmHg with a median use of three (range 0-8) blood pressure lowering drugs. Mean 24-hour blood pressure before RDN was 157/93 (±18/13) mmHg. RDN was performed with different devices, with the Simplicity™ catheter currently used most frequently. CONCLUSION Here we report on the rationale and design of the Dutch RDN registry. Enrolment in this investigator-initiated study is ongoing. We present baseline characteristics of the first 306 participants.


WHO confirms SARS in Chinese journalist.


title: Global health surveillance.

text: Awareness of the importance of global health surveillance increased in the latter part of the 20th century with the global emergence of human immunodeficiency virus and novel strains of influenza. In the first decade of the 21st century, several events further highlighted global shared interests in and vulnerability to infectious diseases. Bioterrorist use of anthrax spores in 2001 raised awareness of the value of public health surveillance for national security. The epidemic of severe acute respiratory syndrome (SARS) in 2003, re-emergence of a panzootic of avian influenza A H5N1 in 2005, and the sudden emergence of pandemic H1N1 in North America in 2009 all highlighted the importance of shared global responsibility for surveillance and disease control. In particular, in 2003, SARS precipitated changes in awareness of the world's collective economic vulnerability to epidemic shocks.


title: Anesthetic management of the child with an upper respiratory tract infection.

text: PURPOSE OF REVIEW The decision to proceed with anesthesia for the child with an upper respiratory tract infection is often difficult. Whereas most studies suggest that children who present for elective procedures with an upper respiratory tract infection are at increased risk of perioperative adverse events, these events are typically easy to recognize and treat. This review will discuss the current literature regarding outcome in children who present for elective surgery with an upper respiratory tract infection and suggests approaches to optimize their perioperative management. RECENT FINDINGS Although the literature regarding this important topic has been slow to evolve, recent large-scale outcome studies have identified a number of factors that increase the risk of perioperative adverse events among children with upper respiratory tract infections. The significance of these findings will be discussed. SUMMARY An understanding of the risk factors associated with administering anesthesia to the child with an upper respiratory tract infection is important in identifying elements of the preoperative assessment that merit attention and in optimizing the anesthetic plan as a means to limit any perioperative complications.


Influenza seasonality: underlying causes and modeling theories.


title: The diagnostic thresholds for synovial fluid analysis in late periprosthetic infection of the hip depend on the duration of symptoms.

text: AIMS We hypothesised that the synovial white blood cell (WBC) count in patients with a late periprosthetic joint infection (PJI) of the hip would depend on the duration of a patient's symptoms, and that the optimal diagnostic threshold would also depend on this period of time. PATIENTS AND METHODS The synovial WBC count and percentage of polymorphonuclear cells (%PMN), and the serum CRP and ESR levels obtained > six weeks after primary THA were compared between 50 infected and 88 non-infected THAs, and in patients with symptoms for more than or less than two weeks. Diagnostic thresholds for the synovial WBC count were calculated using area under the curve calculation. RESULTS The synovial WBC count was significantly higher in patients with symptoms for ≤ two weeks compared with those with symptoms for > two weeks (p = 0.03). The optimal threshold for diagnosing PJI for the synovial WBC count was 5750 cells/µL (sensitivity 94; specificity 100; PPV 100%; NPV 89%; AUC 99%) and 1556 cells/µL (sensitivity 91; specificity 94; PPV 87% and NPV 97%; AUC 95%), respectively. The thresholds for the cut-offs based on duration of symptoms improved the diagnostic performance of this test. CONCLUSION This study shows that the diagnostic thresholds for synovial fluid analysis in late periprosthetic infection following THA may depend on duration of symptoms. Cite this article: Bone Joint J 2016;98-B:1355-9.


title: Pathology of experimental CV777 coronavirus enteritis in piglets. II. Electron microscopic study.

text: Sixteen cesarean-derived, colostrum-deprived piglets were infected oronasally with CV777 coronavirus on the second or third day of life. Two uninfected piglets were controls. After an incubation period of 22 hours to 36 hours, all principals showed severe diarrhea. The piglets were killed at different time intervals. Viral particles were found in the jejunal villous epithelial cells from 18 hours after infection until four days after the beginning of diarrhea. In the colonic epithelial cells, viral particles and degenerative lesions were found only in the piglet killed 36 hours after onset of diarrhea. Degenerative lesions in the enterocytes began at 18 hours after infection and were most pronounced in the jejunum at the onset of clinical signs. From 24 hours on after the onset of clinical signs, three cell types were found: degenerated virus-containing enterocytes; cuboidal cells; and columnar, highly vacuolated cells containing lipid droplets.


title: Experimental analyses of the cavitation generated by ultrasonically activated surgical devices.

text: PURPOSE Recently, the incidence of postoperative pancreatic leakage has been reported to have significantly increased after laparoscopic gastrectomy for gastric cancers in comparison to open surgery. Although their lateral thermal spread has been shown to be smaller than that of other electrosurgical devices, ultrasonically activated surgical devices (USADs) have been suggested as one of the causes. We therefore hypothesized that cavitation generated by USADs could cause injuries to pancreatic tissue. Our retrospective study showed that the amylase activity in the drainage fluid of patients in whom surgery was performed using a USAD with a curved blade was significantly higher than that in patients in whom surgery was performed using a USAD with a straight blade. We therefore focused on the effects of straight and curved USAD blades. METHODS The distribution of microbubbles generated in degassed water, which produce mechanical and biological tissue destructive forces, was measured and compared between the two types of USAD. RESULTS More microbubbles were found to be generated from the side aspect of the curved blade, and the mechanical and biological destructive forces were found to be significantly higher than those generated by the side aspect of the straight blade. CONCLUSIONS These findings strongly suggest that cavitation generated by USADs could injure pancreatic tissues in the clinical cases. Surgeons should be aware of these properties of devices to achieve safe and secure surgeries.


title: Pandemic influenza A(H1N1) 2009: molecular characterisation and duration of viral shedding in intensive care patients in Bordeaux, south-west France, May 2009 to January 2010.

text: From May 2009 to January 2010, the Virology Laboratory at the University Hospital of Bordeaux received more than 4,000 nasopharyngeal samples from the Aquitaine region (south-west France) for the diagnosis of pandemic influenza A(H1N1)2009. Eighty-three infected patients deteriorated and were admitted to intensive care units. Our study focused on 24 of these patients. Positivity for influenza A(H1N1)2009 was monitored by realtime PCR and duration of viral shedding was determined. The first available sample of each patient was analysed for bacterial, fungal and viral co-infection. We observed six bacterial (or bacterial/fungal) co-infections and one viral co-infection with respiratory syncytial virus. The samples were analysed for the presence of the neuraminidase H275Y (N1 numbering) mutation, which confers resistance to oseltamivir, by realtime PCR of the neuraminidase gene. No H275Y mutation was observed in any of the viral strains screened in this study. In parallel, a fragment of the haemagglutinin gene encoding amino acid residues 173 to 362 was sequenced to detect mutations that had been reported to increase the severity of the disease. Two patients were infected by strains bearing the D222G (H3 numbering) mutation. The viral shedding of A(H1N1)2009 in this study ranged from four to 28 days with a median of 11 days.


title: Efficacy of two canine parvovirus vaccines for inducing seroconversion in Rottweiler and Doberman pinscher pups with various levels of maternally derived antibodies.

text: The study reported here investigated the efficacy of two commonly used modified-live virus vaccines to induce seroconversion against canine parvovirus (CPV) in 213 Rottweiler and Doberman pinscher pups with various titers of maternally derived CPV antibody. Beginning at 6 to 8 weeks of age, pups were given a subcutaneous vaccination every 21 days (range, 18-24 days) in the dorsal region of the neck or shoulder area. Pups vaccinated with vaccine A(a) received three vaccinations and completed the vaccination series by 12 to 14 weeks of age. Pups vaccinated with vaccine Bb received four vaccinations and completed the vaccination series by 15 to 17 weeks of age. Antibody titers against CPV in both vaccine groups were similar before vaccination. Pups in the vaccine-A group seroconverted significantly earlier than those in the vaccine-B group. After the first vaccination, more pups with a CPV-2b hemagglutination inhibition (HI) titer of < or = 1:80 responded to vaccine A than to vaccine B. In addition, CPV-2b HI titers after vaccination were also significantly (P < or = 0.05) higher for the pups in the vaccine-A group after first, second, and third vaccinations, compared with those of pups in the vaccine-B group.


One world--one health.


title: Apical entry and release of severe acute respiratory syndrome-associated coronavirus in polarized Calu-3 lung epithelial cells.

text: Severe acute respiratory syndrome (SARS), caused by a novel coronavirus (CoV) known as SARS-CoV, is a contagious and life-threatening respiratory illness with pneumocytes as its main target. A full understanding of how SARS-CoV would interact with lung epithelial cells will be vital for advancing our knowledge of SARS pathogenesis. However, an in vitro model of SARS-CoV infection using relevant lung epithelial cells is not yet available, making it difficult to dissect the pathogenesis of SARS-CoV in the lungs. Here, we report that SARS-CoV can productively infect human bronchial epithelial Calu-3 cells, causing cytopathic effects, a process reflective of its natural course of infection in the lungs. Indirect immunofluorescence studies revealed a preferential expression of angiotensin-converting enzyme 2 (ACE-2), the functional receptor of SARS-CoV, on the apical surface. Importantly, both ACE-2 and viral antigen appeared to preferentially colocalize at the apical domain of infected cells. In highly polarized Calu-3 cells grown on the membrane inserts, we found that cells exposed to virus through the apical rather than the basolateral surface showed high levels of viral replication. Progeny virus was released into the apical chamber at titers up to 5 logs higher than those recovered from the basolateral chambers of polarized cultures. Taken together, these results indicate that SARS-CoV almost exclusively entered and was released from the apical domain of polarized Calu-3 cells, which might provide important insight into the mechanism of transmission and pathogenesis of SARS-CoV.


title: [Ichthyosis vulgaris].

text: This is a review of diagnostic imaging findings in coronavirus disease-2019 (COVID-19). CT findings are frequently bilateral, multilobar and peripheral ground-glass opacities with vascular enlargements. Consolidations often appear during progression as well as crazy paving and reticulation. Lymphadenopathy, pleural effusions and pneumothorax are rare and should raise concern for other disease. Initially the CT can be normal, and the diagnostic performance for COVID-19 is not optimal. However, findings of characteristic ground-glass opacities during the ongoing epidemic should lead to considerations, that the patient may have COVID-19.


title: Role of GNRA motif mutations within stem-loop V of internal ribosome entry segment in coxsackievirus B3 molecular attenuation.

text: The lengthy 5' nontranslated region of coxsackievirus B3 (CVB3) forms a highly ordered secondary structure containing an internal ribosome entry segment (IRES), which plays an important role in controlling viral translation and pathogenesis. The stem-loop V (SL-V) of this IRES contains a large lateral bulge loop which encompasses two conserved GNRA motifs. In this study, we analyzed the effects of point mutations within the GNRA motifs of the CVB3 IRES. We characterized in vitro virus production and translation efficiency and we tested in vivo virulence of two CVB3 mutants produced by site-directed mutagenesis. The GNAA1 and GNAA2 RNAs displayed decreased translation initiation efficiency when translated in rabbit reticulocyte lysates. This translation defect was correlated with reduced yields of infectious virus particles in HeLa cells in comparison with the wild type. When inoculated orally into Swiss mice, both mutant viruses were avirulent and caused neither inflammation nor necrosis in hearts. These results highlight the important role of the GNRA motifs within the SL-V of the IRES of CVB3, in directing translation initiation.


title: Feline coronavirus serotypes 1 and 2: seroprevalence and association with disease in Switzerland.

text: To determine the prevalence of antibodies to feline coronavirus (FCoV) serotypes 1 and 2 in Switzerland and their association with different disease manifestations, a serological study based on immunofluorescence tests was conducted with Swiss field cats using transmissible gastroenteritis virus (TGEV), FCoV type 1 and FCoV type 2 as antigens. A total of 639 serum samples collected in the context of different studies from naturally infected cats were tested. The current study revealed that, with an apparent prevalence of 83%, FCoV serotype 1 is the most prevalent serotype in Switzerland. FCoV type 1 viruses induced higher antibody titers than FCoV type 2, and were more frequently associated with clinical signs and/or feline infectious peritonitis. The antibody development in seven cats experimentally infected with FCoV type 1 revealed that, with progressing duration of infection, antibodies to FCoV type 1 significantly increased over those to FCoV type 2. There was a significant relationship between antibody titers against TGEV, FCoV 1, and FCoV 2 and TGEV antigen detected the highest proportion of seropositive cats. We conclude that a vaccine against FCoV should be based on FCoV type 1-related antigens and that for serodiagnosis of FCoV infection TGEV should be used to attain the highest diagnostic efficiency. When serology is used in addition to clinical signs, hematology, and clinical chemistry results as an aid to diagnose clinical FIP, TGEV shows a diagnostic efficiency equal to that of a FCoV antigen.


title: Dynamics of biological systems: role of systems biology in medical research.

text: Cellular systems are networks of interacting components that change with time in response to external and internal events. Studying the dynamic behavior of these networks is the basis for an understanding of cellular functions and disease mechanisms. Quantitative time-series data leading to meaningful models can improve our knowledge of human physiology in health and disease, and aid the search for earlier diagnoses, better therapies and a healthier life. The advent of systems biology is about to take the leap into clinical research and medical applications. This review emphasizes the importance of a dynamic view and understanding of cell function. We discuss the potential for computer-aided mathematical modeling of biological systems in medical research with examples from some of the major therapeutic areas: cancer, cardiovascular, diabetic and neurodegenerative medicine.


European Resuscitation Council guidelines for resuscitation 2005. Section 2. Adult basic life support and use of automated external defibrillators.


title: The potential anticancer activity of extracts derived from the roots of Scutellaria baicalensis on human oral squamous cell carcinoma cells.

text: Various herb products derived from plants have potent biological effects including anticancer activity. In the present study, the antitumor activity of a herbal product derived from the Scutellaria baicalensis (S. baicalensis) was examined, using in vitro assays in a human oral squamous cell carcinoma (OSCC) cell line. Results showed that S. baicalensis root extract at the concentration of 100 μg/ml inhibited monolayer- and anchorage-independent growth in human OSCC cell lines, while not affecting the adhering abilities of cells. This suggested that it did not alter the expression of any of the adhesion receptors that mediate cell-extracellular matrix (ECM) interactions. The S. baicalensis root extract demonstrated potent cytostatic and apoptotic effects due to the downregulation of the cyclin-dependent kinase 4 expression and its partner cyclin D1, resulting in G1 arrest and poly (ADP-ribose) polymerase (PARP) cleavage. Additionally, the S. baicalensis root extract was found to have blocked vascular endothelial growth factor (VEGF)-induced migration and tube formation in human endothelial cells. Taken together, these results demonstrate that as a herbal product, the S. baicalensis root extract is a potential inhibitor of tumori- and angiogenesis and may be valuable in the development of pharmaceutical medications for the treatment of oral squamous cell carcinoma.


title: Contributions of the structural proteins of severe acute respiratory syndrome coronavirus to protective immunity.

text: We investigated the contributions of the structural proteins of severe acute respiratory syndrome (SARS) coronavirus (CoV) to protective immunity by expressing them individually and in combinations from a recombinant parainfluenza virus (PIV) type 3 vector called BHPIV3. This vector provided direct immunization of the respiratory tract, the major site of SARS transmission, replication, and disease. The BHPIV3/SARS recombinants were evaluated for immunogenicity and protective efficacy in hamsters, which support a high level of pulmonary SARS-CoV replication. A single intranasal administration of BHPIV3 expressing the SARS-CoV spike protein (S) induced a high titer of SARS-CoV-neutralizing serum antibodies, only 2-fold less than that induced by SARS-CoV infection. The expression of S with the two other putative virion envelope proteins, the matrix M and small envelope E proteins, did not augment the neutralizing antibody response. In absence of S, expression of M and E or the nucleocapsid protein N did not induce a detectable serum SARS-CoV-neutralizing antibody response. Immunization with BHPIV3 expressing S provided complete protection against SARS-CoV challenge in the lower respiratory tract and partial protection in the upper respiratory tract. This was augmented slightly by coexpression with M and E. Expression of M, E, or N in the absence of S did not confer detectable protection. These results identify S among the structural proteins as the only significant SARS-CoV neutralization antigen and protective antigen and show that a single mucosal immunization is highly protective in an experimental animal that supports efficient replication of SARS-CoV.


COVID-19 and Its Impact on Nuclear Medicine.


title: [Pharmaceutical care for severe and critically ill patients with COVID-19].

text: Severe and critically ill patients with coronavirus disease 2019 (COVID-19) were usually with underlying diseases, which led to the problems of complicated drug use, potential drug-drug interactions and medication errors in special patients. Based on Diagnosis and treatment of novel coronavirus pneumonia (trial version 6), and Management of COVID-19: the Zhejiang experience, we summarized the experience in the use of antiviral drugs, corticosteroids, vascular active drugs, antibacterial, probiotics, nutrition support schemes in severe and critically ill COVID-19 patients. It is also suggested to focus on medication management for evaluation of drug efficacy and duration of treatment, prevention and treatment of adverse drug reactions, identification of potential drug-drug interactions, individualized medication monitoring based on biosafety protection, and medication administration for special patients.


title: Severe acute respiratory syndrome coronavirus 3C-like proteinase N terminus is indispensable for proteolytic activity but not for enzyme dimerization. Biochemical and thermodynamic investigation in conjunction with molecular dynamics simulations.

text: Severe acute respiratory syndrome (SARS) coronavirus is a novel human coronavirus and is responsible for SARS infection. SARS coronavirus 3C-like proteinase (SARS 3CL(pro)) plays key roles in viral replication and transcription and is an attractive target for anti-SARS drug discovery. In this report, we quantitatively characterized the dimerization features of the full-length and N-terminal residues 1-7 deleted SARS 3CL(pro)s by using glutaraldehyde cross-linking SDS-PAGE, size-exclusion chromatography, and isothermal titration calorimeter techniques. Glutaraldehyde cross-linking SDS-PAGE and size-exclusion chromatography results show that, similar to the full-length SARS 3CL(pro), the N-terminal deleted SARS 3CL(pro) still remains a dimer/monomer mixture within a wide range of protein concentrations. Isothermal titration calorimeter determinations indicate that the equilibrium dissociation constant (K(d)) of the N-terminal deleted proteinase dimer (262 microm) is very similar to that of the full-length proteinase dimer (227 microm). Enzymatic activity assay using the fluorescence resonance energy transfer method reveals that N-terminal deletion results in almost complete loss of enzymatic activity for SARS 3CL(pro). Molecular dynamics and docking simulations demonstrate the N-terminal deleted proteinase dimer adopts a state different from that of the full-length proteinase dimer, which increases the angle between the two protomers and reduces the binding pocket that is not beneficial to the substrate binding. This conclusion is verified by the surface plasmon resonance biosensor determination, indicating that the model substrate cannot bind to the N-terminal deleted proteinase. These results suggest the N terminus is not indispensable for the proteinase dimerization but may fix the dimer at the active state and is therefore vital to enzymatic activity.


title: Particle-associated retroviral RNA and tandem RGH/HERV-W copies on human chromosome 7q: possible components of a 'chain-reaction' triggered by infectious agents in multiple sclerosis?

text: Different groups have observed retrovirus particle (RVP) production in cell cultures from patients with multiple sclerosis (MS). This in vitro production appeared relatively specific for MS versus healthy controls, but was likely to be enhanced or activated by infectious triggers such as Herpesviruses (e.g. HSV, EBV). Independent molecular analysis of retroviral RNA associated with RVP revealed two different genetic families of endogenous retroviral elements (HERV): MSRV/HERV-W and RGH/HERV-H. Interestingly, these sequences were detected by mutually exclusive primers in RT - PCR amplifications. Surprisingly, these two HERV families both contain an ancestral proviral copy inserted in chromosome 7q21-22 region at about 1 kb of distance of each other. Another HERV-W proviral sequence is located within a T-cell alpha-delta receptor (TCR) gene in chromosome 14q11.2 region. Interestingly, these two regions correspond to genetic loci previously identified as potentially associated with 'multigenic' susceptibility to MS and TCR alpha chain genetic determinants have been reported to be statistically associated with MS. A plausible role for infectious agents triggering a co-activation of the chromosome 7q HERV tandem (replicative retrovirus and/or other virus and/or intracellular bacteria) and, eventually, other HERV copies, is discussed. The role of particular HERV polymorphism and the production of pathogenic molecules (gliotoxin and superantigen) possibly associated with retroviral expression are also evoked. An integrative concept of pathogenic 'chain-reaction' in MS involving several step-specific pathogenic 'agents' and 'products' somewhat interacting with particular genetic elements would federate most partial data obtained on MS, including retroviral expression.


title: Cellulose-Chitosan-Keratin Composite Materials: Synthesis, Immunological and Antibacterial Properties.

text: Novel composites were synthesized from keratin (KER), cellulose (CEL) and chitosan (CS). The method is recyclable because majority (>88%) of [BMIm+Cl-], an ionic liquid (IL), used as the sole solvent, was recovered for reuse. Experimentally, it was confirmed that unique properties of each component remain intact in the composites, namely bactericide (from KER and CS) and anti-inflammatory property (from KER). Specifically, the composites were examined for their anti-inflammatory influence on macrophages. The cells were imaged and immunophenotyped to determine the quantity using the macrophage marker CD11b. The 75:25 [KER+CS] composite was found to have the least amount of CD11b macrophages compared to other composites. Bactericidal assays indicated that all composites, except the 25:75 [KER+CS], substantially reduce the growth of organisms such as vancomycin resistant Enterococcus (VRE) and Eschericia coli. The results clearly indicate that the composites possess all properties needed for effective use as a wound dressing.


Pathogenesis of lower respiratory tract infections due to Chlamydia, Mycoplasma, Legionella and viruses.


title: A mouse model for study of systemic HIV-1 infection, antiviral immune responses, and neuroinvasiveness.

text: We created a model of HIV-1 infection of conventional mice for investigation of viral replication, control, and pathogenesis. To target HIV-1 to mice, the coding region of gp120 in HIV-1/NL4-3 was replaced with that of gp80 from ecotropic murine leukemia virus, a retrovirus that infects only rodents. The resulting chimeric virus construct, EcoHIV, productively infected murine lymphocytes, but not human lymphocytes, in culture. Adult, immunocompetent mice were readily susceptible to infection by a single inoculation of EcoHIV as shown by detection of virus in splenic lymphocytes, peritoneal macrophages, and the brain. The virus produced in animals was infectious, as shown by passage in culture, and immunogenic, as shown by induction of antibodies to HIV-1 Gag and Tat. A second chimeric virus based on clade D HIV-1/NDK was also highly infectious in mice; it was detected in both spleen and brain 3 wk after tail vein inoculation, and it induced expression of infection response genes, MCP-1, STAT1, IL-1beta, and complement component C3, in brain tissue as determined by quantitative real-time PCR. EcoHIV infection of mice forms a useful model of HIV-1 infection of human beings for convenient and safe investigation of HIV-1 therapy, vaccines, and potentially pathogenesis.


title: Lipid rafts of primary endothelial cells are essential for Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8-induced phosphatidylinositol 3-kinase and RhoA-GTPases critical for microtubule dynamics and nuclear delivery of viral DNA but dispensable for binding and entry.

text: Early during de novo infection of human microvascular dermal endothelial (HMVEC-d) cells, Kaposi's sarcoma-associated herpesvirus (KSHV) (human herpesvirus 8 [HHV-8]) induces the host cell's preexisting FAK, Src, phosphatidylinositol 3-kinase (PI3-K), Rho-GTPases, Diaphanous-2 (Dia-2), Ezrin, protein kinase C-zeta, extracellular signal-regulated kinase 1/2 (ERK1/2), and NF-kappaB signal pathways that are critical for virus entry, nuclear delivery of viral DNA, and initiation of viral gene expression. Since several of these signal molecules are known to be associated with lipid raft (LR) domains, we investigated the role of LR during KSHV infection of HMVEC-d cells. Pretreatment of cells with LR-disrupting agents methyl beta-cyclo dextrin (MbetaCD) or nystatin significantly inhibited the expression of viral latent (ORF73) and lytic (ORF50) genes. LR disruption did not affect KSHV binding but increased viral DNA internalization. In contrast, association of internalized viral capsids with microtubules (MTs) and the quantity of infected nucleus-associated viral DNA were significantly reduced. Disorganized and disrupted MTs and thick rounded plasma membranes were observed in MbetaCD-treated cells. LR disruption did not affect KSHV-induced FAK and ERK1/2 phosphorylation; in contrast, it increased the phosphorylation of Src, significantly reduced the KSHV-induced PI3-K and RhoA-GTPase and NF-kappaB activation, and reduced the colocalizations of PI3-K and RhoA-GTPase with LRs. Biochemical characterization demonstrated the association of activated PI3-K with LR fractions which was inhibited by MbetaCD treatment. RhoA-GTPase activation was inhibited by PI3-K inhibitors, demonstrating that PI3-K is upstream to RhoA-GTPase. In addition, colocalization of Dia-2, a RhoA-GTPase activated molecule involved in MT activation, with LR was reduced. KSHV-RhoA-GTPase mediated acetylation and aggregation of MTs were also reduced. Taken together, these studies suggest that LRs of endothelial cells play critical roles in KSHV infection and gene expression, probably due to their roles in modulating KSHV-induced PI3-K, RhoA-GTPase, and Dia-2 molecules essential for postbinding and entry stages of infection such as modulation of microtubular dynamics, movement of virus in the cytoplasm, and nuclear delivery of viral DNA.


title: Cross-reaction of SARS-CoV antigen with autoantibodies in autoimmune diseases.

text: To investigate the significance of the SARS-associated coronavirus (SARS-CoV) antibody, detected by ELISA and indirect immunofluorescence assays (IFA) for the SARS-CoV Vero E6 cell lysates, in non-SARS subjects, 114 serum samples from healthy controls and 104 serum specimens from autoimmune disease patients were collected. The results of ELISA showed that among 114 sera from healthy controls, 4 (3.5%) were positive of SARS-CoV-IgG antibody and 114 (100%) were all negative of SARS-CoV-IgM antibody; the specificity of SARS-CoV-IgG antibody for SARS patients was 96.5%, but the specificity of both SARS-CoV-IgG and -IgM antibodies for SARS patients was 100%. In 58 cases with SLE, positive rates of SARS-CoV-IgG and -IgM antibodies were 32.8% (19/58) and 8.6% (5/58), respectively, in which 11 cases (19%) were positive of both SARS-CoV-IgG and -IgM antibodies; in 10 cases with SS, positive rate of both SARS-CoV-IgG and -IgM antibodies was 10% (1/10); in 16 cases with MCTD, positive rate of SARS-CoV-IgG was 37.5% (6/16), positive rate of both SARS-CoV-IgG and -IgM antibodies was 6.3% (1/16); in 20 cases with RA, one case was positive (5%) of SARS-CoV-IgG. However, of all samples with positive SARS-CoV-IgG and -IgM antibodies for autoimmune diseases and healthy controls, SARS-CoV RNA and antibodies were all negative by RT-PCR and IFA. All sera for negative or positive ELISA results were also negative or positive results using ELISA with Vero E6 cells lysates. These studies showed that SARS-CoV Vero E6 cell lysates for the ELISA to detect SARS-CoV antibodies could lead to the false-positive reactions or cross-reactions of SARS-CoV antibodies in non-SARS diseases and healthy controls, and the false-positive reactions or cross-reactions were related to Vero E6 cell lysates and autoantibodies in non-SARS population.


title: [Relationship between acid-base disturbance in blood gas analysis and prognosis of patients with severe acute respiratory syndrome].

text: OBJECTIVE To study the influence of the acute blood gas analysis index change on the prognosis of patients with severe acute respiratory syndrome (SARS). METHODS Artery blood gas analysis indexes were determined in 38 of 103 cases of SARS patients during acute stage, the relationship between the change of blood gas analysis index and prognosis of patients with SARS was analyzed. RESULTS The lower partical pressure of carbon dioxide in artery (PaCO2), partical pressure of oxygen in artery (PaO2) in blood was, the higher death rate was. pH in blood had no significant influence on the prognosis of SARS patients. The prognosis of SARS patients were worse and the death rate was higher when patient had two or more acid-base disturbance. CONCLUSION The artery blood gas analysis is useful for judging the conditions and prognosis of patients with SARS, and is beneficial for clinical treatment.


title: Intranasal interferon as protection against experimental respiratory coronavirus infection in volunteers.

text: In a double-blind, placebo-controlled study, self-administered intranasal human interferon alpha A produced by recombinant DNA technology was given both before and after virus challenge with a respiratory coronavirus. The incidence of colds, the severity of symptoms and signs, and virus replication were all reduced in subjects receiving interferon as compared with those given placebo.


title: Subcellular localization of the severe acute respiratory syndrome coronavirus nucleocapsid protein.

text: The coronavirus nucleocapsid (N) protein is a viral RNA-binding protein with multiple functions in terms of virus replication and modulating cell signalling pathways. N protein is composed of three distinct regions containing RNA-binding motif(s), and appropriate signals for modulating cell signalling. The subcellular localization of severe acute respiratory syndrome coronavirus (SARS-CoV) N protein was studied. In infected cells, SARS-CoV N protein localized exclusively to the cytoplasm. In contrast to the avian coronavirus N protein, overexpressed SARS-CoV N protein remained principally localized to the cytoplasm, with very few cells exhibiting nucleolar localization. Bioinformatic analysis and deletion mutagenesis coupled to confocal microscopy and live-cell imaging, revealed that SARS-CoV N protein regions I and III contained nuclear localization signals and region II contained a nucleolar retention signal. However, cytoplasmic localization was directed by region III and was the dominant localization signal in the protein.


title: Transmission potential of primary pneumonic plague: time inhomogeneous evaluation based on historical documents of the transmission network.

text: BACKGROUND The transmission potential of primary pneumonic plague, caused by Yersinia pestis, is one of the key epidemiological determinants of a potential biological weapon, and requires clarification and time dependent interpretation. METHOD This study estimated the reproduction number and its time dependent change through investigations of outbreaks in Mukden, China (1946), and Madagascar (1957). Reconstruction of an epidemic tree, which shows who infected whom, from the observed dates of onset was performed using the serial interval. Furthermore, a likelihood based approach was used for the time inhomogeneous evaluation of the outbreaks for which there was scarcity of cases. RESULTS According to the estimates, the basic reproduction number, R(0), was on the order of 2.8 to 3.5, which is higher than previous estimates. The lower 95% confidence intervals of R(0) exceeded unity. The effective reproduction number declined below unity after control measures were introduced in Mukden, and before the official implementation in Madagascar. CONCLUSION While the time course of the latter outbreak could be explained by intrinsic factors and stochasticity in this remote and scarcely populated area, the former in Mukden suggests the possible continued chains of transmission in highly populated areas. Using the proposed methods, the who infected whom information permitted the evaluation of the time inhomogeneous transmission potential in relation to public health measures. The study also tackles the problem of statistical estimation of R(0) based on similar information, which was previously performed simply by counting the number of secondary transmissions regardless of time.


title: Identification of severe acute respiratory syndrome coronavirus replicase products and characterization of papain-like protease activity.

text: Gene 1 of the coronavirus associated with severe acute respiratory syndrome (SARS) encodes replicase polyproteins that are predicted to be processed into 16 nonstructural proteins (nsps 1 to 16) by two viral proteases, a papain-like protease (PLpro) and a 3C-like protease (3CLpro). Here, we identify SARS coronavirus amino-terminal replicase products nsp1, nsp2, and nsp3 and describe trans-cleavage assays that characterize the protease activity required to generate these products. We generated polyclonal antisera to glutathione S-transferase-replicase fusion proteins and used the antisera to detect replicase intermediates and products in pulse-chase experiments. We found that nsp1 (p20) is rapidly processed from the replicase polyprotein. In contrast, processing at the nsp2/3 site is less efficient, since a approximately 300-kDa intermediate (NSP2-3) is detected, but ultimately nsp2 (p71) and nsp3 (p213) are generated. We found that SARS coronavirus replicase products can be detected by 4 h postinfection in the cytoplasm of infected cells and that nsps 1 to 3 colocalize with newly synthesized viral RNA in punctate, perinuclear sites consistent with their predicted role in viral RNA synthesis. To determine if PLpro is responsible for processing these products, we cloned and expressed the PLpro domain and the predicted substrates and established PLpro trans-cleavage assays. We found that the PLpro domain is sufficient for processing the predicted nsp1/2 and nsp2/3 sites. Interestingly, expression of an extended region of PLpro that includes the downstream hydrophobic domain was required for processing at the predicted nsp3/4 site. We found that the hydrophobic domain is inserted into membranes and that the lumenal domain is glycosylated at asparagine residues 2249 and 2252. Thus, the hydrophobic domain may anchor the replication complex to intracellular membranes. These studies revealed that PLpro can cleave in trans at the three predicted cleavage sites and that it requires membrane association to process the nsp3/4 cleavage site.


title: [Effectiveness of personal protective measures in prevention of nosocomial transmission of severe acute respiratory syndrome].

text: OBJECTIVE To evaluate the effectiveness of personal protective measures of health care workers (HCWs) against severe acute respiratory syndrome (SARS). METHODS A case-control study from ten hospitals in Guangdong, with 180 non-infected and 77 infected staff members that accessed the isolation unit every day, and participated in direct first aid for severe SARS patients. All participants were surveyed about how they were using personal protective equipment (PPE), protective drugs and hygiene habits when caring for patients with SARS. Statistical analysis was done with either chi(2) or Fisher's exact test for univariate analysis, whereas we used forward stepwise selection (Waldesian) for logistic regression. RESULTS Univariate analysis showed that mask, gown, gloves, goggles, footwear, "hand-washing and disinfecting", gargle, "membrane protection", "taking shower and changing clothing after work", "avoid from eating and drinking in ward", oseltamivir phospha tall had protective effects (P < 0.05), but stepwise logistic regression showed significant differences for mask (OR = 0.78, 95% CI: 0.60 - 0.99), goggles (OR = 0.20, 95% CI: 0.10 - 0.41) and footwear (OR = 0.58, 95% CI: 0.39 - 0.86). Analysis for linear trend in proportions showed that dose response relationship existed in mask, gown, gloves, goggles, footwear, gargle, "membrane protection" and "taking shower and changing dree after work" (P < 0.01). The attack rate of HCWs who were rescuing severe SARS patients without any PPE was 61.5% (16/26). It seemed that the more the protective measures were used, the higher the protective effect was (P < 0.001), and could reach 100% if mask, gown, gloves, goggles, footwear, "hand-washing and disinfecting" were all used at the same time. CONCLUSIONS Nosocomial infection of SARS can be prevented effectively by precautions against droplets and personal contact. HCWs must take strict protection according to the guidance of WHO or Chinese MOH and pay attention to personal hygiene.


title: Decline in prevalence and asymmetric distribution of human T cell lymphotropic virus 1 and 2 in blood donors, State of Minas Gerais, Brazil, 1993 to 2007.

text: INTRODUCTION Human T cell lymphotropic virus types 1 and 2 (HTLV-1/2) are endemic in Brazil and are screened for in transfusion services since 1993. This study evaluated the evolution of the prevalence of HTLV-1 and 2 in blood donors of the Hemominas Foundation from 1993 to 2007, and its geographical distribution in State of Minas Gerais, Brazil. METHODS The Hemominas Foundation is a centralized blood center in Minas Gerais, Brazil. The sources of data were the Hemominas Foundation Technical Bulletin and files from the centralized serological laboratory. Donors were tested in the period using enzyme linked immuno sorbent assays (ELISA), followed by Western blot, when repeatedly reactive. The data were analyzed by EPIINFO 6.2 and TABWIN 3.5 softwares. RESULTS The average seroprevalence in the period 1993-2007 was 0.1%. A steady decline occurred from 0.4% in 1993 to below 0.1% in 2002 and later, with a transient peak of 0.5% in 1994. HTLV reactivity distribution was asymmetrical in the state, with regions of higher prevalence, interspersed with low prevalence areas. Comparison of positive and negative donors verified that increasing age was proportional to virus positivity. Odds ratio for age ranged from 1.43 (30 to 39 years-old) to 3.09 (50 to 65 years-old). Women had a greater chance of being positive (OR-1.64), as previously described. CONCLUSIONS Possible explanations for HTLV-1/2 prevalence decline are the exclusion of positive donors from the donor pool, an increase in repeat donors and ELISA test improvement, with reduction in the number of false positive results.


title: Structures of two coronavirus main proteases: implications for substrate binding and antiviral drug design.

text: Coronaviruses (CoVs) can infect humans and multiple species of animals, causing a wide spectrum of diseases. The coronavirus main protease (M(pro)), which plays a pivotal role in viral gene expression and replication through the proteolytic processing of replicase polyproteins, is an attractive target for anti-CoV drug design. In this study, the crystal structures of infectious bronchitis virus (IBV) M(pro) and a severe acute respiratory syndrome CoV (SARS-CoV) M(pro) mutant (H41A), in complex with an N-terminal autocleavage substrate, were individually determined to elucidate the structural flexibility and substrate binding of M(pro). A monomeric form of IBV M(pro) was identified for the first time in CoV M(pro) structures. A comparison of these two structures to other available M(pro) structures provides new insights for the design of substrate-based inhibitors targeting CoV M(pro)s. Furthermore, a Michael acceptor inhibitor (named N3) was cocrystallized with IBV M(pro) and was found to demonstrate in vitro inactivation of IBV M(pro) and potent antiviral activity against IBV in chicken embryos. This provides a feasible animal model for designing wide-spectrum inhibitors against CoV-associated diseases. The structure-based optimization of N3 has yielded two more efficacious lead compounds, N27 and H16, with potent inhibition against SARS-CoV M(pro).


title: Bioterrorism and catastrophe response: a quick-reference guide to resources.

text: BACKGROUND Dentists' responses to catastrophe have been redefined by bioterrorism. Informed response requires accurate information about agents and diseases that have the potential to be used as weapons. METHODS The authors reviewed information about the most probable bioterrorist weapons (those from the Center for Disease Control and Prevention's Category A) from the World Wide Web and print journals and distilled it into a resource list that is current, relevant to dentistry and noncommercial. The Web sites cited include those sponsored by federal agencies, academic institutions and professional organizations. The articles cited include those published in English within the last six years in refereed journals that are available in most higher education institutions. RESULTS The authors present the information in a table that provides a quick-reference guide to resources describing agents and diseases with the greatest potential for use as weapons: anthrax, botulism, plague, smallpox, tularemia and viral hemorrhagic fevers. This article presents Web site and journal citations for background and patient-oriented information (fact sheets), signs and symptoms, and prophylactic measures and treatment for each of the agents and diseases. The table facilitates quick access to this information, especially in an emergency. This article also points out guidelines for response should a suspected attack occur. CONCLUSIONS Armed with information about biological weapons, dentists can provide faster diagnosis, inform their patients about risks, prophylaxis or treatment and rethink their own role in terrorism response. CLINICAL IMPLICATIONS Fast, accurate diagnosis limits the spread of exceptionally contagious diseases. Providing accurate information to patients minimizes misinformation and the associated public fear and panic that, unchecked, could overwhelm health care systems.


[Myositis].


Myositis.


title: Extremely low exposure of a community to severe acute respiratory syndrome coronavirus: false seropositivity due to use of bacterially derived antigens.

text: Estimates of seropositivity to a new infectious agent in a community are useful to public health. For severe acute respiratory syndrome (SARS), the figures are conflicting. Herein, we screened 12,000 people in a community stricken by SARS 10 months previously and found 53 individuals (0.44%) who had immunoglobulin G antibodies to the SARS coronavirus (SARS-CoV) nucleocapsid (N) produced in bacteria. However, only seven of these (group 1) had sera which also reacted with the native N antigen expressed in SARS-CoV-infected Vero cells, N-transfected 293T cells, and tissues of infected SARS patients. Of these, six individuals had had SARS previously. The remaining person, as well as the 46 other individuals (group 2), were healthy and had no history of SARS. Group 1 antibodies recognized epitopes located slightly differently in N from those of group 2 antibodies, and a mouse hybridoma antibody resembling the former type was generated. Unusually, group 2 antibodies appeared to recognize cross-reactive bacterial epitopes that presumably were posttranslationally modified in eukaryotes and hence were probably not induced by SARS-CoV or related coronaviruses but rather by bacteria. The N antigen is thus highly unique. The extremely low rate (0.008%) of asymptomatic SARS infection found attests to the high virulence of the SARS-CoV virus.


title: Prevalence, clinical presentations and complications among hospitalized children with influenza pneumonia.

text: Viral respiratory tract infections are a major cause of hospitalization in children. Influenza is common but often not laboratory proven. We report a prospective study of children admitted with a clinical diagnosis of pneumonia. Infants and children (ages 1 month-15 years) who were hospitalized with community-acquired pneumonia were enrolled in the study. Their nasopharyngeal aspirated samples were analyzed for common respiratory viruses, including influenza virus, by reverse transcription-polymerase chain reaction (RT-PCR) or PCR. Out of 257 patients, we identified 127 (49.4%) cases with respiratory viruses, and influenza was found in 32 of these cases (12.5%). Other common respiratory viruses included respiratory syncytial virus in 42 (16.3%), human metapneumovirus in 24 (9.3%), adenovirus in 17 (6.6%) and parainfluenza virus in 12 (4.7%). The median age of the influenza group was 2 years and 3 months, and 27 (84%) of children in this group were under the age of 5. Asthma was the most common co-morbidity (4/32, 12.5%). Common clinical presentations were fever and cough (100%) with crepitations (90%). The median length of hospitalization was 6 days. Three patients developed respiratory failure, with one mortality (3.1%). One child developed infection-associated hemophagocytic syndrome. Our study demonstrated that young children had a high risk of hospitalization due to influenza pneumonia, which contributed to a significant morbidity.


Viral infections of the liver in childhood.


title: Preparation and characterization of monoclonal antibodies against S1 domain at N-terminal residues 249 to 667 of SARS-associated coronavirus S1 protein.

text: OBJECTIVE To prepare and characterize monoclonal antibodies (mAbs) against S1 protein of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV). METHODS 6-His-tagged recombinant fragment at N-terminal residues 249 to 667 of SARS-CoV S1 protein including S-protein receptor-binding domain was expressed in E.coli. The immunogenicity of this S1 domain was identified and used to immunize BALB/c mice for the production of hybridomas. The identification of the mAbs against this S1 domain was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA) and Western blotting, respectively. RESULTS Three hybridomas producing mAbs specific to the S1 domain was obtained, with a relative molecular mass of 48,500. None of the 3 mAbs were reactive with human coronaviruses 229E and OC43. Two of the mAbs were IgG2a isotype, and the other was IgG1. CONCLUSIONS This is the first report of mAbs produced against S-protein receptor-binding domain of SARS-CoV. The 3 S1-specific mAbs may be useful for further study of the function of the S protein and for diagnosis of SARS-CoV infection.


title: Induction of autoreactive CD8+ cytotoxic T cells during Theiler's murine encephalomyelitis virus infection: implications for autoimmunity.

text: Theiler's murine encephalomyelitis virus (TMEV) belongs to the family Picornaviridae and causes demyelinating disease in the spinal cords of infected mice. Although immune responses have been shown to play an important role in demyelination, the precise effector mechanism(s) is unknown. Potentially autoreactive cytotoxic cells could contribute to the destruction. We tested whether an autoreactive cell induced by TMEV infection mediated cytotoxicity by using a 5-h (51)Cr release assay in SJL/J mice. Spleen cells from TMEV-infected mice were stimulated with irradiated TMEV antigen-presenting cells and used as effector cells. The effector cells differed from conventional cytotoxic T cells since these cells could kill both TMEV-infected and uninfected syngeneic or semisyngenic cell lines (PSJLSV and BxSF11gSV) but could not kill an allogeneic cell line (C57SV). The TMEV-induced autoreactive cells were also different from conventional natural killer (NK) cells or lymphokine-activated killer (LAK) cells, because they could kill neither NK cell-sensitive YAC-1 nor NK cell-resistant P815 and EL4 cells. Induction of autoreactive cells was not detected in vaccinia virus infection. The autoreactive killing required direct cell-to-cell contact and was mediated by a Fas-FasL pathway but not by a perforin pathway. The phenotype of the killer cells was CD3(+) CD4(-) CD8(+). Intracerebral inoculation of the effector cells into naive mice caused meningitis and perivascular cuffing not only in the brain parenchyma but also in the spinal cord, with no evidence of viral antigen-positive cells. This is the first report demonstrating that TMEV can induce autoreactive cytotoxic cells that induce central nervous system pathology.


title: Prevalence of Cryptosporidium sp in equids in Louisiana.

text: In 1985, 22 pony foals reared in a helminth-free environment were tested daily for oocysts of Cryptosporidium sp by use of fecal flotation. Oocysts were found in all foals. Oocysts were first observed in feces collected from foals 9 to 28 days after birth. The mean period of oocyst shedding was 10 days and ranged from 2 to 18 days in individual foals. Diarrhea was observed in 14 of 22 (64%) foals and began before the period of oocyst shedding. Fecal samples also were examined for other infective agents. Salmonella poona was isolated from 1 foal that did not have diarrhea, and coronavirus particles were observed in the feces of 2 foals with diarrhea. Cryptosporidium sp oocysts also were observed in feces of 2 of 17 Thoroughbred foals, 3 of 14 Quarter Horse foals, and 3 of 26 pony foals reared on pastures with their dams. Samples from pasture-reared foals were collected at irregular intervals. Of the 11 Cryptosporidium-positive fecal samples collected from pastured foals, 2 were from foals with diarrhea. A similar survey was conducted during the 1986 foaling season, using the same procedures. Examination of 300 samples from 58 Quarter Horse, Arabian, and pony foals did not detect oocysts. Daily examination of feces from 10 pony foals reared under helminth-free conditions for 30 days also failed to detect Cryptosporidium oocysts.


title: Current and future therapeutic approaches to the common cold.

text: For decades, investigators have strived to elucidate the pathogenesis, and hence a treatment, for the common cold. Therapy has been tried with a variety of agents, ranging from anecdotal folk remedies, to well-designed medications. Measures primarily directed to relieve the symptoms of the common cold, rather than specific antimicrobial agents, are the current main stay of therapy. Millions of patients would benefit from an easy-to-perform diagnostic test and specific therapy that works quickly, shortening the duration of illness and preventing further spread of infection.


title: The potential impact of climate change on infectious diseases of Arctic fauna.

text: Climate change is already affecting Arctic species including infectious disease agents and greater changes are expected. Some infectious diseases are already increasing but future changes are difficult to predict because of the complexity of host-agent-environment relationships. However mechanisms related to climate change that will influence disease patterns are understood. Warmer temperatures will benefit free living bacteria and parasites whose survival and development is limited by temperature. Warmer temperatures could promote survivability, shorter development rates and transmission. Insects such as mosquitoes and ticks that transmit disease agents may also benefit from climate change as well as the diseases they spread. Climate change will have significant impacts on biodiversity. Disease agents of species that benefit from warming will likely become more prevalent. Host species stressed by changing environmental conditions may be more vulnerable to disease agents. Warming could lead to increased agriculture and other economic opportunities in the Arctic bringing people, domestic food animals, pets and invasive species and their disease agents into Northern regions. Climate warming may also favor the release of persistent environmental pollutants some of which can affect the immune system and may favor increased rates of some diseases.


Public Health Laboratories and the Affordable Care Act: What the New Health-Care System Means for Public Health Preparedness.


title: Bioinformatic approaches for modeling the substrate specificity of HIV-1 protease: an overview.

text: HIV-1 protease has a broad and complex substrate specificity, which hitherto has escaped a simple comprehensive definition. This, and the relatively high mutation rate of the retroviral protease, makes it challenging to design effective protease inhibitors. Several attempts have been made during the last two decades to elucidate the enigmatic cleavage specificity of HIV-1 protease and to predict cleavage of novel substrates using bioinformatic analysis methods. This review describes the methods that have been utilized to date to address this important problem and the results achieved. The data sets used are also reviewed and important aspects of these are highlighted.


title: [SARS-CoV infection and pregnancy].

text: SARS is a highly contagious infection, caused by new coronavirus SARS-CoV. Immunopathological mechanisms responsible for the reaction to SARS-CoV infection have not yet been fully elucidated. Cytokine profile of SARS patients showed marked elevation of Th1 cytokine, interferon gamma, inflammatory cytokines for at least 2 weeks after the onset of the disease. The clinical manifestation of SARS in patients has been of varied nature. Fever of more then 38 degrees C, lasting more then 24 hours, is the most frequently encountered symptom. Other symptoms are non specific and they may include: sore throat, myalgia and nausea. The results of the radiological investigation may appear normal. Infants born to pregnant women with SARS did not appear to have acquired the infection through vertical transmission. However, direct contact with the maternal body fluid which contained SARS-CoV, has put the infants in great danger of perinatal infection. Ribavirin and corticosteroids are usually suggested for the treatment of SARS. However, the ribavirin therapy increases the risk of teratogenic effects in newborns of pregnant women with SARS. Therefore, the usage of this drug is not recommended during pregnancy and lactation.


title: Nucleotide sequence and expression of the spike (S) gene of canine coronavirus and comparison with the S proteins of feline and porcine coronaviruses.

text: We have cloned, sequenced and expressed the spike (S) gene of canine coronavirus (CCV; strain K378). Its deduced amino acid sequence has revealed features in common with other coronavirus S proteins: a stretch of hydrophobic amino acids at the amino terminus (the putative signal sequence), another hydrophobic region at the carboxy terminus (the membrane anchor), heptad repeats preceding the anchor, and a cysteine-rich region located just downstream from it. Like other representatives of the same antigenic cluster (CCV-Insavc-1 strain, feline infectious peritonitis and enteric coronaviruses, porcine transmissible gastroenteritis and respiratory coronaviruses, and the human coronavirus HCV 229E), the CCV S polypeptide lacks a proteolytic cleavage site present in many other coronavirus S proteins. Pairwise comparisons of the S amino acid sequences within the antigenic cluster demonstrated that the two CCV strains (K378 and Insavc-1) are 93.3% identical, about as similar to each other as they are to the two feline coronaviruses. The porcine sequences are clearly more divergent mainly due to the large differences in the amino-terminal (residues 1 to 300) domains of the proteins; when only the carboxy-terminal parts (residues 301 and on) are considered the homologies between the canine, feline and porcine S polypeptides are generally quite high, with identities ranging from 90.8% to 96.8% . The human coronavirus is less related to the other members of the antigenic group. A phylogenetic tree constructed on the basis of the S sequences showed that the two CCVs are evolutionarily more related to the feline than to the porcine viruses. Expression of the CCV S gene using the vaccinia virus T7 RNA polymerase system yielded a protein of the expected M(r) (approximately 200K) which could be immunoprecipitated with an anti-feline infectious peritonitis virus polyclonal serum and which was indistinguishable from the S protein synthesized in CCV-infected cells.


title: SARS-CoV nucleocapsid protein induced apoptosis of COS-1 mediated by the mitochondrial pathway.

text: To investigate the apoptosis effect of SARS coronavirus nucleocapsid protein on cultured cell lines and to explore the possible pathway of apoptosis. pCDNA3.1(-)/his-myc vector containing the SARS coronavirus nucleocapsid gene (N), matric gene (M), spike gene (S) were transfected into COS-1, Huh-7 and HepG2 cells. Apoptosis induced by SARS coronavirus N protein under starvation of serum of COS-1 cells was monitored by Annexin V and electron microscopy assays. Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (DeltaPsim) were determined by flow cytometric assay. Cytochrome C, cleaved caspase (cysteine aspartic acid protease)-3, 9, and poly (ADP-ribose) polymerase (PARP) were detected by Western blot. After removal of serum in COS-1 cells, we observed the loss of DeltaPsim, the increase of ROS and cytochrome C release into cytosol and subsequent activation of caspase-3 and PARP cleavage. The pan-caspase inhibitor z-VAD-fmk can block the activation of caspase 3, 9 and PARP cleavage. In conclusion, SARS coronavirus N protein can induce apoptosis of COS-1 cells by activating mitochondrial pathway. SARS coronavirus M, S protein can not induce apoptosis in COS-1, HepG2 and Huh-7 and SARS coronavirus N protein can not induce apoptosis in HepG2 and Huh-7 by methods used in this study.


title: [Protective effect of specific antibody in serum of convalescent patient with SARS].

text: OBJECTIVE To investigate inhibitory effect of serum severe acute respiratory syndrome (SARS) -specific antibodies from convalescent patients after half an year of onset on SARS-CoV-mediated cytopathic response. METHODS SARS-CoV immunoglobulin G (IgG) antibody was determined by enzyme linked immunoadsorbent assay (ELISA). Twelve serum samples from convalescent patients, diluted by 1:8 with maintenance medium, were mixed with the three dilution supernatants of SARS-CoV. SARS-CoV were isolated, cultured and identified by the Guangzhou Institute of Respiratory Disease, and cultured with Vero E6 cell suspension. The extent of cytopathic response was observed. RESULTS The absorbance (A) value of SARS-CoV IgG antibody ranged from 0.81 to 2.06 in patients after half an year of SARS onset, and form 0.79 to 2.01 in patients before half an year of SARS onset. The extent of cytopathic response was decreased by more than 25% in all 12 convalescent patients, as compared with control serum. CONCLUSION The A value of SARS-CoV IgG antibody in serum of convalescent patients tended to elevate in half an year after SARS onset. SARS-CoV IgG antibody could inhibit SARS-CoV-mediated cytopathic response, indicating it might be one of protective antibodies.


title: The development, optimization and validation of an assay for high throughput antiviral drug screening against Dengue virus.

text: Dengue virus (DENV) is listed as one of the NIAID Category A priority pathogens. Dengue disease is endemic in most tropical countries, with an estimated 2.5 billion people living in areas at risk of DENV infection. Due to the lack of vaccines and antiviral drugs, it is now a huge public health burden around the world. In order to screen large compound libraries for the identification of novel antivirals targeting DENV, it is essential to develop a high throughput screening (HTS) amenable assay. Here, we present the development, optimization and validation of a cytopathic effect-based assay against Dengue virus serotype-2 (DENV-2). The assay conditions, including cell culturing conditions, DMSO tolerance and the multiplicity of infection, were optimized in both 96- and 384-well plates. Assay robustness and reproducibility were determined under the optimized conditions in 96-well plate, including Z'-value of 0.71, signal-to-background ratio of 6.88, coefficient of variation of 6.3% in mock-infected cells and 12.3% in DENV-2 infected cells. This assay was further miniaturized into a 384-well plate format with similar assay robustness and reproducibility comparing with these in the 96-well plate format. This assay was then validated using the LOPAC(1280) compound library, demonstrating its repeatability with comparable assay robustness and reproducibility. This fully developed and validated HTS amenable assay could be used in future studies to screen large compound libraries for the identification of novel antivirals against dengue disease.


title: Use of a reverse-transcriptase polymerase chain reaction for monitoring the shedding of feline coronavirus by healthy cats.

text: The pattern of shedding of feline coronavirus (FCoV) was established in 155 naturally infected pet cats from 29 households over periods of up to five years. Viral RNA was detected in faeces by reverse-transcriptase PCR (RT-PCR), and plasma antiviral antibodies by immunofluorescence. The cats rarely shed FCoV in their saliva. Three patterns of FCoV shedding were observed. Eighteen of the cats shed virus continuously, so were persistent, and possibly lifelong, carriers; none of them developed feline infectious peritonitis. Fifty-six cats ceased shedding virus, although they were susceptible to reinfection, and 44 shed intermittently or were being continuously reinfected. Four of the cats were resistant to infection. Seventy-three per cent of the virus shedding episodes lasted up to three months and 95 per cent up to nine months. There was a correlation between shedding and antibody titre but the cats could remain seropositive for some time after they had ceased shedding virus. One-off testing for FCoV by RT-PCR is inappropriate. Identification of longterm carriers requires that a positive result be obtained by RT-PCR on faecal samples for at least eight consecutive months. A cat should be shown to be negative over five months, or to have become seronegative, to ensure that it has ceased shedding virus.


title: Neurological manifestations in severe acute respiratory syndrome.

text: During the worldwide outbreak of severe acute respiratory syndrome (SARS) in 2002-2003, there were 664 probable SARS patients reported in Taiwan. SARS patients usually present with symptoms related to the respiratory system while neurological manifestations have rarely been described. There were three patients who developed axonopathic polyneuropathy 3-4 weeks after onset of SARS; their clinical condition and electrophysiological studies revealed obvious improvement at follow-up. Two SARS patients have experienced myopathy and three other patients developed rhabdomyolysis. These neuromuscular disorders in SARS patients were considered as critical illness neuropathy and myopathy, but the possibility of direct attack by SARS coronavirus on the nerve and muscle could not be excluded. Large artery ischemic stroke were described in five SARS patients with poor prognosis. Multiple factors contributed to this vascular insult included hypercoagulabe status related to both SARS coronavirous and the usage of intravenous immunoglobulin, septic and cardiogenic shock, and possible vasculitis. The relationship between SARS and above neurological problems still needs further clarification. Pathological and microbiological studies are mandatory to delineate this issue.


title: Palpable Fear: COVID-19's Toll on Physician Morale and Mental Health.

text: As COVID-19 ravaged human lives, it also took a toll on physician morale and mental health.


title: Protection studies on winter dysentery caused by bovine coronavirus in cattle using antigens prepared from infected cell lysates.

text: Cells infected with bovine coronavirus (BCV) were solubilized with Triton X-100 to yield a cell lysate (CL) antigen having high hemagglutinating (HA) titers. The antigen gave high HA titers using rat erythrocytes, suggesting that it contained large amounts of hemagglutinin esterase (HE) antigen. The CL antigen, combined with an oil adjuvant, was tested for protective and antibody-inducing activities in cattle. Four groups (2 cattle/group) of cattle were inoculated with CL antigen having HA titers of 16 000, 4000, 1000, and 250. Another group served as untreated controls. Two intramuscular inoculations were given at an interval of 3 wk. The animals were challenged with virus 1 wk after the second inoculation. The groups immunized with the CL antigen having an HA titer of 4000 or 16 000 produced hemagglutination inhibition (HI) antibody titers of > 320 and serum neutralizing (SN) antibody titers of > 1280. These groups of animals showed no clinical abnormalities after challenge. In the groups immunized with CL antigen at an HA titer of 1000 or 250, HI antibody titers were 40 to 160 and SN titers were 80 to 640. The cattle with HI antibody titers of > or = 160 and the SN titers of > or = 640 showed no clinical signs, but the cattle with the HI antibody titer < 80 and the SN antibody titer < 160 developed watery diarrhea and fever after challenge. These results indicate that CL antigen with high HA titer induces antibody production in cattle that provides effective protection against winter dysentery.


title: Retrospective serological investigation of severe acute respiratory syndrome coronavirus antibodies in recruits from mainland China.

text: Different assays were used to analyze 1,621 serum specimens collected from military recruits from the People's Republic of China in 2002 for severe acute respiratory syndrome (SARS) coronavirus antibodies. The results demonstrated that the subjects either had rarely been exposed to the virus before the 2003 SARS outbreak or had not been exposed but the nucleocapsid protein cross-reacted with other antibodies in humans.


title: Protein nanopatterns and biosensors using gold binding polypeptide as a fusion partner.

text: An efficient strategy for immobilizing proteins on a gold surface was developed by employing the gold binding polypeptide (GBP) as a fusion partner. Using the enhanced green fluorescent protein (EGFP), severe acute respiratory syndrome coronavirus (SARS-CoV) envelope protein (SCVme), and core streptavidin (cSA) of Streptomyces avidinii as model proteins, specific immobilization of the GBP-fusion proteins onto the gold nanoparticles and generation of protein nanopatterns on the bare gold surface were demonstrated. The GBP-fused SCVme bound to gold nanoparticles successfully interacted with its antibody and showed changes in absorbance and color, allowing efficient diagnosis of SARS-CoV. The fusion proteins could be successfully immobilized on the gold surface by nanopatterning and microcontact printing as examined by atomic force microscopy and surface plasmon resonance analysis. The poly(dimethylsiloxane) microfluidic channels were created on the gold surface and were used for antigen-antibody and DNA-DNA interaction studies. Specific immobilization of GBP-EGFP fusion protein and its interaction with the antibody in the microchannels could be demonstrated. By immobilizing the DNA probe through the use of GBP-fused cSA, specific hybridization of the target DNA prepared from Salmonella could also be achieved. The GBP-fusion method allows immobilization of proteins onto the gold surface without surface modification and in bioactive forms suitable for studying protein-protein, DNA-DNA, and other biomolecular interaction studies. Furthermore, these studies can be carried out in a microfluidic system, which allows high-throughput analysis of biomolecular interactions.


title: Life-threatening human metapneumovirus pneumonia requiring extracorporeal membrane oxygenation in a preterm infant.

text: We present the first report in the literature of a child with human metapneumovirus pneumonia who required extracorporeal membrane oxygenation for survival. This was a 3-month-old premature boy from British Columbia, Canada, who developed severe respiratory failure, experienced failure of high-frequency oscillatory mechanical ventilation, and required extracorporeal membrane oxygenation support for 10 days. This case illustrates the importance of including this newly discovered pathogen among the causes of childhood pneumonia.


title: [SARS-CoV-2 infection and solid organ transplantation].

text: Transplantation has become a valid therapeutic option for an increasing number of patients with end-stage organ disease. The emergence of SARS-CoV-2 coronavirus infection and associated disease (COVID-19) has alarmed the transplant community, since recommendations for adequate follow-up of organ transplant recipients during the acute phase of a pandemic are limited. Furthermore, treatment options against COVID-19 disease and adequate adjustment of immunosuppression in at risk patients remain a concern. This review summarizes current knowledge on the incidence and clinical course of SARS-CoV-2 infection in patients with solid organ transplantation. It also discusses therapeutic strategies and provides general recommendations on how to proceed with transplantation programs in a time when health care resources may become scarce.


title: Incidence and surgical treatment of synthetic mesh-related infectious complications after laparoscopic ventral rectopexy.

text: BACKGROUND Prosthetic-related infection and erosion occurring after a laparoscopic ventral rectopexy (LVR) are rare complications, and their importance is often underestimated. The aim of this study was to compare the incidence rate and surgical management of these complications in LVR patients with polyester (PE) or polypropylene (PP) prostheses. METHODS From January 2004 to June 2012, 149 patients underwent LVR with PE and 176 underwent LVR with PP. Surgical management and rate of infectious and erosive prosthesis-related complications, depending on the type of prosthesis, were described and compared. Functional results after complications were assessed. RESULTS Five patients from the PE prosthesis group (3.3 %), compared with two patients from the PP prosthesis group (1.1 %), experienced prosthesis-related infection or erosion (p = 0.16). The rate of erosion alone was 3.3 % in patients with a PE prosthesis, and 0.55 % in patients with a PP prosthesis (p = 0.06). The average time until clinical diagnosis of a prosthesis-related complication was identical for both groups: 31 months (range 3-62 months). All patients underwent surgical removal of the prosthesis: For the five patients from the PE group, complete removal was performed by laparoscopy associated with a transanal procedure. For the two patients in the PP mesh group, laparoscopy was ineffective in removing the mesh which was partially removed through a subsequent transanal procedure. None of the patients had a protective stoma, and in all patients the complication had resolved 12 months after removal. Only one patient had worsening functional symptoms (fecal incontinence) after prosthesis removal. CONCLUSIONS When a prosthesis-related infection or erosion occurs, treatment consists in the surgical removal of the prosthesis by laparoscopy/and/or a transanal procedure. Functional symptoms do not routinely recur after prosthesis removal.


title: Identification of the coronavirus MHV-JHM mRNA 4 product.

text: A bacterial expression vector was constructed to encode a fusion protein which had, at its carboxy terminus, a polypeptide encoded within the 5' proximal open reading frame of the coronavirus MHV-JHM mRNA 4. This polypeptide was isolated and used to produce an antiserum. The antiserum reacted specifically with a 15,000 Mr polypeptide synthesized in MHV-JHM-infected cells, or in vitro translations of infected cell poly(A) RNA enriched for mRNA 4. These results demonstrate the translational activity of mRNA 4 during infection, identify conclusively the translation product and provide a means to investigate the synthesis and function of this protein.


Global impact of severe acute respiratory syndrome: measures to prevent importation into Saudi Arabia.


Protein microarrays.


The Hong Kong SARS report: an Australian perspective.


title: Transoral stapling for Zenker diverticulum: effect of the traction suture-assisted technique on long-term outcomes.

text: BACKGROUND Long-term outcomes and predictors of success after transoral stapling for Zenker diverticulum are still unclear. METHODS Between 2001 and 2010, 91 patients with Zenker diverticulum underwent transoral stapling under general anesthesia. Since 2008, the technique was modified by applying traction sutures to ease engagement of the common septum inside the stapler jaws. Perioperative variables, distribution of symptoms, and outcome of surgery were analyzed. Long-term results were compared between patients undergoing standard versus modified technique of transoral stapling. RESULTS The transoral approach was successfully completed in 79 (86.8 %) patients with a median age of 74 years. Overall morbidity was 5 %, and there was no mortality. The median length of hospital stay was 2 days. Six patients were lost to follow-up. After a median follow-up of 53 (range, 12-114) months, an improvement of dysphagia and regurgitation scores (p < 0.001) and a reduction in the number of pneumonia episodes per year (p < 0.001) was recorded. The long-term success rate of the procedure was 80.1 %. At a median time of 12 months, 14 patients complained of recurrent symptoms, 7 of whom needed an open (n = 4) or transoral (n = 3) reoperation. Use of traction sutures resulted in a greater long-term success compared with the standard procedure (p = 0.04). CONCLUSIONS Transoral stapling is a safe and effective technique. A repeat procedure is feasible in case of recurrent diverticulum. The use of traction sutures applied at the apex of the common septum before stapling might increase the long-term success of the technique.


title: Artificial Intelligence-Powered Search Tools and Resources in the Fight Against COVID-19.

text: Emerging technologies are set to play an important role in our response to the COVID-19 pandemic. This paper explores three prominent initiatives: COVID-19 focused datasets (e.g., CORD-19); Artificial intelligence-powered search tools (e.g., WellAI, SciSight); and contact tracing based on mobile communication technology. We believe that increasing awareness of these tools will be important in future research into the disease, COVID-19, and the virus, SARS-CoV-2.


title: [Characterization of human coronavirus 229E infection among patients with respiratory symptom in Beijing, Oct-Dec, 2007].

text: OBJECTIVE To know the etiology, prevalence, clinical symptoms associated with the infection of the HCoV-229E in the respiratory specimens sampled from adult patients in Beijing. METHODS 158 nasopharyngeal swab specimens were collected from adult patients with fever in Beijing between October and December, 2007. We performed the screening of HCoV-229E by real-time RT-PCR and sequencing of HCoV-229E gene fragments derived from conventional PCR. At meantime, we also screened the HCoV-229E positive samples for the co-infection with HCoV-NL63, HCoV-HKU1 and HMPV by real-time RT-PCR. Finally, demographic and clinical data associated with HCoV-229E infection were examined retrospectively. RESULTS We detected 103 (62.5%) of 158 specimens were positive for HCoV-229E by real-time RT-PCR. When tested for other respiratory viruses, 26 HCoV-229E positive patients were found to be co-infected with other viruses. Of which HCoV-NL63 was observed in 3 specimens (11.5%), HCoV-HKU1 in 3 (11.5%) and HMPV in 20 (76.9%). The main clinical manifestations were noted as: headache (in 70.9%), sore throat (69%), chills (68%), cough (33%), sputum (21.3%), rhinorrhea (21.4%), nasal obstruction (16.5%), and a few of patients were visible as vomiting (6.8%), dyspnea (3.9%), diarrhea (in 1.9%). The rate of HCoV-229E infection in adult patients was found no relative with age and gender. CONCLUSION Our data showed that HCoV-229E is a common and important pathogen in adult patients with acute respiratory symptoms but usually resulted in milder influenza-like illnesses. There might have a local outbreak of HCoV-229E infection in Beijing, Oct-Dec, 2007.


Emerging diseases fuel health screening.


title: Severe acute respiratory syndrome: radiographic appearances and pattern of progression in 138 patients.

text: PURPOSE To retrospectively evaluate the radiographic appearances and pattern of progression of severe acute respiratory syndrome (SARS). MATERIALS AND METHODS Chest radiographs obtained at clinical presentation and during treatment in 138 patients with confirmed SARS (66 men, 72 women; mean age, 39 years; age range, 20-83 years) were assessed. Radiographic appearances of pulmonary parenchymal abnormality, distribution, and extent of involvement on initial chest radiographs were documented. Recognizable patterns of radiographic progression were determined by comparing the overall mean percentage of lung involvement for each patient on serial radiographs. RESULTS Initial chest radiographs were abnormal in 108 of 138 (78.3%) patients and showed air-space opacity. Lower lung zone (70 of 108, 64.8%) and right lung (82 of 108, 75.9%) were more commonly involved. In most patients, peripheral lung involvement was more common (81 of 108, 75.0%). Unifocal involvement (59 of 108, 54.6%) was more common than multifocal or bilateral involvement. No cavitation, lymphadenopathy, or pleural effusion was demonstrated. Four patterns of radiographic progression were recognized: type 1 (initial radiographic deterioration to peak level followed by radiographic improvement) in 97 of 138 patients (70.3%), type 2 (fluctuating radiographic changes) in 24 patients (17.4%), type 3 (static radiographic appearance) in 10 patients (7.3%), and type 4 (progressive radiographic deterioration) in seven patients (5.1%). Initial focal air-space opacity in 44 of 59 patients (74.6%) progressed to unilateral multifocal or bilateral involvement during treatment. CONCLUSION Predominant peripheral location; common progression pattern from unilateral focal air-space opacity to unilateral multifocal or bilateral involvement during treatment; and lack of cavitation, lymphadenopathy, and pleural effusion are the more distinctive radiographic findings of SARS.


title: [Two genotypes of Canine coronavirus simultaneously detected in the fecal samples of healthy foxes and raccoon dogs].

text: 61 fecal samples from healthy foxes and 24 from healthy raccoon dogs were examined by RT-nested PCR assays for the presence and genotypic identification of Canine coronaviruses (CCVs). 77.0% fox samples were recognized as CCV positive, 43 of which belonged to type II and 29 to type I, as well as both genotypes were simultaneously detected in 25 samples. Out of the total 24 fecal samples from raccoon dogs, 22 were CCV positive for type II and 16 for type I. M gene fragments of 8 samples were sequenced, 4 of which were confirmed as CCV type I and the other 4 as CCV type II. Sequence analysis showed that the M gene of CCV type I had a high similarity of 96.7% - 98.1% between the fox-and raccoon dog strains and the reported Italian strain from diarrhea dogs. The two genotypes, with an identity of 88.3% - 89.7%, formed two separate branches in phylogenetic tree. Interestingly, the sequence at several nucleic acid sites of CCV type II differed between foxes and raccoon dogs. The co-existence and popularity of the two CCV genotypes in healthy foxes and raccoon dogs were first confirmed in this article.