title: Antigenic and immunogenic characterization of recombinant baculovirus-expressed severe acute respiratory syndrome coronavirus spike protein: implication for vaccine design.

text: The spike (S) glycoprotein of severe acute respiratory syndrome coronavirus (SARS-CoV) mediates the receptor interaction and immune recognition and is considered a major target for vaccine design. However, its antigenic and immunogenic properties remain to be elucidated. In this study, we immunized mice with full-length S protein (FL-S) or its extracellular domain (EC-S) expressed by recombinant baculoviruses in insect cells. We found that the immunized mice developed high titers of anti-S antibodies with potent neutralizing activities against SARS pseudoviruses constructed with the S proteins of Tor2, GD03T13, and SZ3, the representative strains of 2002 to 2003 and 2003 to 2004 human SARS-CoV and palm civet SARS-CoV, respectively. These data suggest that the recombinant baculovirus-expressed S protein vaccines possess excellent immunogenicity, thereby inducing highly potent neutralizing responses against human and animal SARS-CoV variants. The antigenic structure of the S protein was characterized by a panel of 38 monoclonal antibodies (MAbs) isolated from the immunized mice. The epitopes of most anti-S MAbs (32 of 38) were localized within the S1 domain, and those of the remaining 6 MAbs were mapped to the S2 domain. Among the anti-S1 MAbs, 17 MAbs targeted the N-terminal region (amino acids [aa] 12 to 327), 9 MAbs recognized the receptor-binding domain (RBD; aa 318 to 510), and 6 MAbs reacted with the C-terminal region of S1 domain that contains the major immunodominant site (aa 528 to 635). Strikingly, all of the RBD-specific MAbs had potent neutralizing activity, 6 of which efficiently blocked the receptor binding, confirming that the RBD contains the main neutralizing epitopes and that blockage of the receptor association is the major mechanism of SARS-CoV neutralization. Five MAbs specific for the S1 N-terminal region exhibited moderate neutralizing activity, but none of the MAbs reacting with the S2 domain and the major immunodominant site in S1 showed neutralizing activity. All of the neutralizing MAbs recognize conformational epitopes. These data provide important information for understanding the antigenicity and immunogenicity of S protein and for designing SARS vaccines. This panel of anti-S MAbs can be used as tools for studying the structure and function of the SARS-CoV S protein.


title: Putative cis-acting stem-loops in the 5' untranslated region of the severe acute respiratory syndrome coronavirus can substitute for their mouse hepatitis virus counterparts.

text: Consensus covariation-based secondary structural models for the 5' 140 nucleotides of the 5' untranslated regions (5'UTRs) from mouse hepatitis virus (MHV) and severe acute respiratory syndrome coronavirus (SCoV) were developed and predicted three major helical stem-loop structures, designated stem-loop 1 (SL1), SL2, and SL4. The SCoV 5'UTR was predicted to contain a fourth stem-loop, named SL3, in which the leader transcriptional regulatory sequence (TRS) is folded into a hairpin loop. cDNAs corresponding to MHV/SCoV chimeric genomes were constructed by replacing the complete MHV 5'UTR with the corresponding SCoV sequence and by separately replacing MHV 5'UTR putative SL1, putative SL2, TRS, and putative SL4 with the corresponding SCoV sequences. Chimeric genomes were transcribed in vitro, and viruses were recovered after electroporation into permissive cells. Genomes in which the MHV 5'UTR SL1, SL2, and SL4 were individually replaced by their SCoV counterparts were viable. Chimeras containing the complete SCoV 5'UTR or the predicted SCoV SL3 were not viable. A chimera containing the SCoV 5'UTR in which the SCoV TRS was replaced with the MHV TRS was also not viable. The chimera containing the entire SCoV 5'UTR failed to direct the synthesis of any virus-specific RNA. Replacing the SCoV TRS with the MHV TRS in the MHV/5'UTR SCoV chimera permitted the synthesis of minus-sense genome-sized RNA but did not support the production of positive- or minus-sense subgenomic RNA7. A similar phenotype was obtained with the MHV/SCoV SL3 chimera. These results suggest a role for the TRS in the replication of minus-sense genomic RNA in addition to its known function in subgenomic RNA synthesis.


title: Direct Fmoc/tert-Bu solid phase synthesis of octamannosyl polylysine dendrimer-peptide conjugates.

text: The mannose binding proteins on the surface of the dendritic cells are responsible for capture of pathogens in the early stages of immune response. Conjugation to mannose dendrimers is a rarely explored but potentially powerful strategy for enhancing immunogenicity of synthetic peptides relying on direct delivery to dendritic cells. We describe a general protocol for preparation of pure, monodisperse third-generation mannosylated poly-L-lysine dendrimer-peptide conjugates using direct, machine-assisted Fmoc/t-Bu solid phase peptide synthesis. The glycodendrons were elaborated onto the N- or C-terminus of sequences derived from HIV-1 gp41, SARS-CoV S2 protein, and Influenza Hemagglutinin (consisting of 15-44 residues). The products were obtained in a homogeneous state after cleavage from the resin, deprotection, and a single purification on semipreparative RP-HPLC.


title: [Hypothesis on generating and tracer gas study regarding transmission of severe acute respiratory syndrome through ventilation system in a general hospital].

text: OBJECTIVE By measuring airflow and ventilation distribution of ward building, to explore and verify the hypothesis of airborne transmission and risk factor of severe acute respiratory syndrome (SARS) nosocomial infection. METHODS Tracer gas (perfume of plant oil) was emitted to the bathroom of wards when SARS index patient lived. Six different experimental situations were designed to control the status of exhaust fan in bathrooms, exhaust fan in the top of building and fresh air exchange system. The concentration of perfume was separately measured by 4 groups of lab workers and recorded blindly by the scores of "tenth degree". RESULTS Tracer gas was detected from the wards of 8th to 13th floor. CONCLUSION Architecture and ventilation system of the inpatient building in the hospital contributed to the aerodynamic condition of SARS nosocomial infection through airborne transmission. The distribution of tracer gas in the wards was associated with SARS patients in this building. It was possible that SARS could have been transmitted to for distance by aerosol or other carriers.


title: [Role of siRNAS and miRNAS in the processes of RNA-mediated gene silencing during viral infections].

text: Phenomenon of RNA-induced gene silencing is a highly conservative mechanism among eukaryotic organisms. Several classes of small RNAs (siRNAs and miRNAs) 21-25 nt in length, which play a significant role in the processes of development of an organism, occurred important components of antiviral defence in animals and plants. This review shortly describes the main stages of gene silencing mechanism, features of antiviral RNA silencing in plants, invertebrates, mammals, ways of suppression of RNA-interference by viruses, as well as possible approaches of utilization of abovementioned phenomenon for struggling against viral infections.


title: Emerging coronaviruses: first SARS, second MERS and third SARS-CoV-2: epidemiological updates of COVID-19.

text: Since December 2019, the emergence of the Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) infection has been reported unexpectedly in Wuhan, China, with staggering infection speed across China and around the world. To date, seven known strains of HCoVs belonging to four genera (i.e., α?, β?, γ, and δ-CoV) have been recognized; the latest one has been identified as the SARS-CoV-2. Although the common transmission routes of SARS-CoV-2 is the respiratory tract, it seems that other routes such as the gastrointestinal tract may be effective for the entry of the virus in the body. Although there are no biological markers to predict the susceptibility of humans to COVID-19, several risk factors have been identified to predict the susceptibility of patients to COVID-19. Initial data revealed that males, pregnant women, elderly, and underlying conditions predispose patients to higher morbidity or mortality and also might be at risk for a severe infection of COVID-19. There is a greater need to better understand the mechanisms and risk factors of transmission routes. To date, despite the whole world effort to review various aspects of SARS-CoV-2, including epidemiology, clinical manifestations, diagnosis, and treatment options, there are still gaps in the knowledge of this disease and many issues remain unclear. Therefore, there is an urgent need for update data on SARS-CoV-2. Here, this study provide the current epidemiological status (transmission routes and risk of transmission, possible origins and source, mortality and morbidity risk, and geographical distribution) of the SARS-CoV-2 in the world in 2020.


title: Global Health Governance at a Crossroads.

text: This review takes stock of the global health governance (GHG) literature. We address the transition from international health governance (IHG) to global health governance, identify major actors, and explain some challenges and successes in GHG. We analyze the framing of health as national security, human security, human rights, and global public good, and the implications of these various frames. We also establish and examine from the literature GHG's major themes and issues, which include: 1) persistent GHG problems; 2) different approaches to tackling health challenges (vertical, horizontal, and diagonal); 3) health's multisectoral connections; 4) neoliberalism and the global economy; 5) the framing of health (e.g. as a security issue, as a foreign policy issue, as a human rights issue, and as a global public good); 6) global health inequalities; 7) local and country ownership and capacity; 8) international law in GHG; and 9) research gaps in GHG. We find that decades-old challenges in GHG persist and GHG needs a new way forward. A framework called shared health governance offers promise.


DNA vaccines: a safe and efficient platform technology for responding to emerging infectious diseases.


title: Bovine cryptosporidiosis in Denmark. 2. Cryptosporidia associated with neonatal calf diarrhea.

text: In recent years cryptosporidia have often been identified in diarrheic calves of under one month old, either as the sole pathogenic agent or in combination with other diarrhea-causing infectious agents. In 203 cases of bovine cryptosporidiosis, recorded over a period of two years, cryptosporidia were present in such great numbers that they would seem to be of causal significance. The calves were from three days to five weeks old. In 111 cases (55%) a monoinfection with cryptosporidia occurred, while in the rest of the cases mixed infections with other agents associated with calf diarrhea were demonstrated, especially rotavirus (61 cases) and coronavirus (32 cases); in addition, mixed infections with ETEC (four cases), septicaemic E. coli (two cases), salmonellosis (11 cases), and BVD and coccidiosis (one case each) were seen. In 87 (10%) of 849 4-21-day-old dead calves a massive occurrence of cryptosporidia was demonstrated. Extreme emaciation was noted in 36 of 122 autopsied calves. Most of these calves were more than 14 days old, and mono-infection with cryptosporidia was demonstrated in two thirds of them. In younger calves, more acute cases of cryptosporidiosis were commoner, often in connection with concomitant infections with rota- or coronavirus.


Coronaviruses and multiple sclerosis.


title: A is for aphorisms - feed a fever, starve a cold? Or could it be starve a fever, feed a cold?

text: The recommendations for management of fever have changed throughout the course of history, as has the very concept of fever. It has changed from something viewed as a diagnosis in its own right to a sign of something to be measured and as a trigger for a diagnostic trail to be made.


title: Pandemic influenza planning in nursing homes: are we prepared?

text: Avian influenza or Influenza A (H5N1) is caused by a viral strain that occurs naturally in wild birds, but to which humans are immunologically naïve. If an influenza pandemic occurs, it is expected to have dire consequences, including millions of deaths, social disruption, and enormous economic consequences. The Department of Health and Human Resources plan, released in November 2005, clearly affirms the threat of a pandemic. Anticipating a disruption in many factions of society, every segment of the healthcare industry, including nursing homes, will be affected and will need to be self-sufficient. Disruption of vaccine distribution during the seasonal influenza vaccine shortage during the 2004/05 influenza season is but one example of erratic emergency planning. Nursing homes will have to make vital decisions and provide care to older adults who will not be on the initial priority list for vaccine. At the same time, nursing homes will face an anticipated shortage of antiviral medications and be expected to provide surge capacity for overwhelmed hospitals. This article provides an overview of current recommendations for pandemic preparedness and the potential effect of a pandemic on the nursing home industry. It highlights the need for collaborative planning and dialogue between nursing homes and various stakeholders already heavily invested in pandemic preparedness.


title: Severe acute respiratory syndrome: public health response and clinical practice update for an emerging disease.

text: PURPOSE OF REVIEW Severe acute respiratory syndrome is an emerging infectious disease with high morbidity and mortality and potential for global spread. This review highlights the salient points of the 2002-2003 outbreak to help clinicians with early recognition, treatment, disease control, and prevention. RECENT FINDINGS Severe acute respiratory syndrome is a respiratory illness caused by a novel coronavirus, severe acute respiratory syndrome-coronavirus. The disease was first recognized in Asia in February 2003 and, over the next several months, spread to more than two dozen countries in North and South America, Europe, and Asia. The disease is characterized by fever with symptomatic and radiographic evidence of pneumonia. SUMMARY Because diagnosis cannot be established by clinical findings alone, epidemiologic findings (including history of travel to affected areas, close contact with other case patients, or linkage to clusters of unexplained pneumonia) play a critical role in the diagnostic evaluation. Infection control procedures (including droplet and airborne precautions) are critical for preventing transmission.


title: When family-centered care is challenged by infectious disease: pediatric health care delivery during the SARS outbreaks.

text: In this ethnographic study, the authors examined the experiences and perspectives of children hospitalized because of SARS (severe acute respiratory syndrome), their parents, and pediatric health care providers. The sample included 5 children, 10 parents, and 8 health care providers who were directly affected by SARS during the time of the outbreaks and extreme infection control procedures. The data analyses illuminated a range of perceived experiences for this triadic sample. Issues related to social isolation due to infection control precautions were predominant. Themes included emotional upheaval, communication challenges, and changes in parental and professional roles. These findings reveal the cogent effects of SARS on family-centered care. The notion of providing family-centered care within an environment plagued by an infectious outbreak suggests an omniously difficult task. Efforts must be made to optimize family-centered care despite obstacles. The authors suggest effective clinical approaches in the event of future outbreaks.


title: Analyses of Merging Clinical and Viral Genetic Data for Influenza Surveillance.

text: The annual influenza vaccine is one of the most common public health interventions and is universally recommended for all individuals older than six months. Vaccine composition depends on viruses circulating over the past flu season and are estimated to be the most prevalent and representative strains in the current season. Here, we use clinical data outfitted with viral genetics to characterize confirmed influenza cases from the past two flu seasons and genetically compare them to the strains that they were vaccinated against that year. We show that case similarities to vaccine strains differ by geographic region and that the vaccines appear to have different levels of effectiveness by region. This study demonstrates the value of merging viral genetics with clinical data. Further research is needed to formally evaluate whether this improves biosurveillance efforts and enhances efficacy of influenza vaccines.


title: Analysis of the aphthovirus 2A/2B polyprotein 'cleavage' mechanism indicates not a proteolytic reaction, but a novel translational effect: a putative ribosomal 'skip'.

text: The 2A region of the aphthovirus foot-and-mouth disease virus (FMDV) polyprotein is only 18 aa long. A 'primary' intramolecular polyprotein processing event mediated by 2A occurs at its own C terminus. FMDV 2A activity was studied in artificial polyproteins in which sequences encoding reporter proteins flanked the 2A sequence such that a single, long, open reading frame was created. The self-processing properties of these artificial polyproteins were investigated and the co-translational 'cleavage' products quantified. The processing products from our artificial polyprotein systems showed a molar excess of 'cleavage' product N-terminal of 2A over the product C-terminal of 2A. A series of experiments was performed to characterize our in vitro translation systems. These experiments eliminated the translational or transcriptional properties of the in vitro systems as an explanation for this imbalance. In addition, the processing products derived from a control construct encoding the P1P2 region of the human rhinovirus polyprotein, known to be proteolytically processed, were quantified and found to be equimolar. Translation of a construct encoding green fluorescent protein (GFP), FMDV 2A and beta-glucuronidase, also in a single open reading frame, in the presence of puromycin, showed this antibiotic to be preferentially incorporated into the [GFP2A] translation product. We conclude that the discrete translation products from our artificial polyproteins are not produced by proteolysis. We propose that the FMDV 2A sequence, rather than representing a proteolytic element, modifies the activity of the ribosome to promote hydrolysis of the peptidyl(2A)-tRNA(Gly) ester linkage, thereby releasing the polypeptide from the translational complex, in a manner that allows the synthesis of a discrete downstream translation product to proceed. This process produces a ribosomal 'skip' from one codon to the next without the formation of a peptide bond.


title: Molecular diagnosis of animal diseases: some experiences over the past decade.

text: The experiences of a veterinary research and diagnostic laboratory are summarized on the development and application of the PCR to diagnose a wide range of viral diseases in animals. The group started the routine diagnostic application of the PCR as early as 1988 and today a total of 35 nested PCR assays are in routine use for the detection of 15 DNA and 20 RNA viruses. Special tools and laboratory practice were applied to avoid false-positive results, while false-negatives are avoided by internal controls (mimics). At present, the classical nested PCR methods are being replaced by real-time TaqMan and molecular beacon assays and the multiplex real-time PCR detection of viruses is also under development. By direct sequencing of the PCR products, phylogeny studies are performed and molecular epizootiology results are provided for rapid and exact identification of virus variants. Molecular epizootiology also contributes to trace the routes of virus spreading on large geographic areas. Recently, large efforts have been made to follow the recommendations of Office International des Epizooties for the standardization and international harmonization of the molecular diagnostic assays.


title: [Etiological structure of acute respiratory viral infections morbidity in Novosibirsk and Novosibirsk region in epidemic season 2011-2012].

text: INTRODUCTION ARI occupying the first place in the structure of total human morbidity. The aim of the study was to investigate the species diversity of the viruses causing AR among residents of the Novosibirsk region during epidemic season (October to April). MATERIALS AND METHODS 164 nasopharyngeal swabs were collected and analyzed. Viral RNA/DNA, cDNA synthesis and PCR were carried out employing "RIBO-prep" "eReverta-L", "AmpliSens Influenza virus A/B-FL" and "AmpliSens ARI-screen-FL" kits (CRI of Epidemiology). RESULTS Etiological agent of the disease was found in 69(43%) samples. Monoinfection was found in 58 (35%). In 14 (9%) samples were detected serogroup I coronaviruses, in 13 (8%) rhinoviruses, in 7 (4%) respiratory syncytial virus, in 6 (4%) parainfluenza virus type 1, in 5 (3%) parainfluenza virus type 3. Adenoviruses and bocavirus were identified in 3 (2%) samples. Parainfluenza virus type 2 and 4, metapneumovirus, serogroup Il coronaviruses (HKU1 and OC43) were presented in 2 (1%) samples. In 11 (7%) samples was found mixed infection. CONCLUSION The majority of common colds were caused by serogroup I coronaviruses (NL63 and 229E), rhinoviruses and mixed infections. The peak of species variability of viruses caused acute respiratory infections was determined in age group of children 2-4 years old. In older age groups the species variability of analyzed viruses was decreased, rhinovirus infection becomes prevalent.


title: Purification and crystallization reveal two types of interactions of the fusion protein homotrimer of Semliki Forest virus.

text: The fusion proteins of the alphaviruses and flaviviruses have a similar native structure and convert to a highly stable homotrimer conformation during the fusion of the viral and target membranes. The properties of the alpha- and flavivirus fusion proteins distinguish them from the class I viral fusion proteins, such as influenza virus hemagglutinin, and establish them as the first members of the class II fusion proteins. Understanding how this new class carries out membrane fusion will require analysis of the structural basis for both the interaction of the protein subunits within the homotrimer and their interaction with the viral and target membranes. To this end we report a purification method for the E1 ectodomain homotrimer from the alphavirus Semliki Forest virus. The purified protein is trimeric, detergent soluble, retains the characteristic stability of the starting homotrimer, and is free of lipid and other contaminants. In contrast to the postfusion structures that have been determined for the class I proteins, the E1 homotrimer contains the fusion peptide region responsible for interaction with target membranes. This E1 trimer preparation is an excellent candidate for structural studies of the class II viral fusion proteins, and we report conditions that generate three-dimensional crystals suitable for analysis by X-ray diffraction. Determination of the structure will provide our first high-resolution views of both the low-pH-induced trimeric conformation and the target membrane-interacting region of the alphavirus fusion protein.


Characterization and location of the structural polypeptides of turkey enteric coronavirus using monoclonal antibodies and enzymatic treatments.


title: Antigenic relationships among proteins of bovine coronavirus, human respiratory coronavirus OC43, and mouse hepatitis coronavirus A59.

text: Antisera prepared against each of three single and one pair of major structural proteins of the bovine coronavirus (Mebus strain) were used in immunoblotting studies to measure cross-reactivity with the structural proteins of the human coronavirus OC43 and the mouse hepatitis coronavirus A59. We conclude that the bovine coronavirus is comprised of four major structural proteins, gp190 (normally present as 120- and 100-kilodalton subunits), gp140, pp52, and gp26. The human coronavirus OC43 has an antigenically homologous counterpart of similar molecular mass to each of these proteins. The mouse hepatitis coronavirus A59 has an antigenically homologous counterpart to only three of these proteins: gp190, pp52 and gp26. There is no counterpart in the mouse virus to the 140-kilodalton glycoprotein, the apparent hemagglutinin of the bovine coronavirus.


Social justice is a global issue: ethical pandemic planning.


title: An ancient role for nuclear beta-catenin in the evolution of axial polarity and germ layer segregation.

text: The human oncogene beta-catenin is a bifunctional protein with critical roles in both cell adhesion and transcriptional regulation in the Wnt pathway. Wnt/beta-catenin signalling has been implicated in developmental processes as diverse as elaboration of embryonic polarity, formation of germ layers, neural patterning, spindle orientation and gap junction communication, but the ancestral function of beta-catenin remains unclear. In many animal embryos, activation of beta-catenin signalling occurs in blastomeres that mark the site of gastrulation and endomesoderm formation, raising the possibility that asymmetric activation of beta-catenin signalling specified embryonic polarity and segregated germ layers in the common ancestor of bilaterally symmetrical animals. To test whether nuclear translocation of beta-catenin is involved in axial identity and/or germ layer formation in 'pre-bilaterians', we examined the in vivo distribution, stability and function of beta-catenin protein in embryos of the sea anemone Nematostella vectensis (Cnidaria, Anthozoa). Here we show that N. vectensis beta-catenin is differentially stabilized along the oral-aboral axis, translocated into nuclei in cells at the site of gastrulation and used to specify entoderm, indicating an evolutionarily ancient role for this protein in early pattern formation.


title: Infections and asthma.

text: Infections have a variety of influences in the asthmatic patient. Respiratory syncytial virus (RSV) is the most common cause of wheezing illnesses in children under the age of 2 years and may be a risk factor for the inception of asthma in the first decade of life. Once asthma has become recognized clinically, rhinovirus is the leading cause of acute asthma exacerbations in adults and older children. Certain respiratory infections may induce chronic lower airway inflammation that may contribute to disease progression or severity in asthma. Paradoxically, some infections may even protect against the development of asthma. This article reviews the evidence for these associations and the mechanisms proposed to explain these various outcomes.


title: Inter-laboratory comparison of the CD-1 neonatal mouse logistic dose-response model for Cryptosporidium parvum oocysts.

text: cryptosporidium parvum oocyst viability can be determined by vital dyes, in vitro excystation, and cell culture; however, neonatal mouse infectivity assays are the reference method. Unfortunately, there have been few efforts to standardize methods for infectivity assays thus casting a veil of uncertainty over the significance and comparability of results. In order to address this issue, two laboratories proficient in measuring oocyst infectivity conducted independent dose titration studies with neonatal CD-1 mice using standardized protocols and a well-characterized isolate of Cryptosporidium parvum. The resulting independent logistic dose-response models derived by regression analysis were compared with each other and with a published model. The comparisons showed these dose-response functions to be reproducible under standardized conditions. It is important to standardize mouse strain, age of mice at inoculation and necropsy, oocyst isolate, and age of oocysts. However, other factors, including methods used to detect infectivity and to count oocyst doses, appear less critical. Adopting a standardized assay for oocyst infectivity will provide both a basis for comparing data from various oocyst disinfection studies and a suitable platform for evaluating new or existing in vitro viability surrogates such as excystation, vital dyes or cell culture.


title: Central Aspects of Nausea and Vomiting in GI Disorders.

text: OPINION STATEMENT Nausea and vomiting result from continuous interactions among gastrointestinal, central nervous system, and autonomic nervous system. Despite being closely associated, central pathways of nausea and vomiting appear to be at least partly different and nausea is no longer considered only a penultimate step of vomiting. Although our understanding of central pathways of nausea has improved over the last one decade, it is still very basic. Afferent pathways from gastrointestinal tract via vagus, vestibular system, and chemoreceptor trigger zone project to nucleus tractus solitarius which, in turn, relays the signal to central pattern generator initiating multiple downstream pathways. This central nausea pathway appears to be under constant modulation by autonomic nervous system and cerebral cortex. There is also some evidence that central pathway of chronic nausea is different from that of acute nausea and closely resembles that of neuropathic pain. This improved understanding has modified the way we can approach the treatment of acute and chronic nausea. While conventional therapies such as antiemetics (antiserotoninergic, antihistaminic, antidopaminergic) and prokinetics are commonly used to manage acute nausea, they are not as effective in improving chronic nausea. Recently, neuromodulators such as tricyclic antidepressants, gabapentin, olanzapine, benzodiazepines, and cannabinoids have been shown to have antinausea effect. There is a need to study the utility of these drugs in managing chronic functional nausea. Improving our understanding of central and peripheral circuitry of nausea will allow us to better utilize the currently available drugs and develop new therapeutic options.


title: Plague: from natural disease to bioterrorism.

text: Yersinia pestis is the causative agent of plague, an enzootic vectorborne disease usually infecting rodents (rats) and fleas. Humans can become infected after being bitten by fleas that have fed on infected rodents. In humans, the disease usually occurs in the form of bubonic plague. In rare cases, the infection spreads to the lungs via the bloodstream and causes secondary pneumonic plague. Person-to-person transmission has been described for pneumonic plague but is rare in primary bubonic plague. Bubonic plague can usually be treated successfully with antibiotics; however, pneumonic plague develops rapidly and carries a high fatality rate despite immediate treatment with antibiotics. Plague is also recognized as a potential agent of bioterrorism. It has been used, or considered for use, as a biologic weapon on several occasions. It is important for the medical community to be familiar with the epidemiology, diagnosis, and symptoms of plague so it can deliver an appropriate and calm response should the unthinkable happen.


title: Case report of the first Severe Acute Respiratory Syndrome patient in China: successful application of extracorporeal liver support MARS therapy in multiorgan failure possibly induced by Severe Acute Respiratory Syndrome.

text: A previously healthy patient was transferred to our infectious department with a 9-day-history of continued fever. The patient was placed on assisted respiration support in addition to anti-viral medication. The diagnosis of SARS (Severe Acute Respiratory Syndrome) was made in view of the severe hypoxemia and the characteristic symptoms exhibited by the patient. Despite the best intensive therapy, he clinically deteriorated into multiorgan dysfunction syndrome (MODS) including additional dysfunction of kidney, liver, and heart. We initiated MARS therapy (extracorporeal liver support utilizing albumin dialysis) with intention to positively influence the organ functions in his MODS on the basis of recently published studies which suggested a positive impact of MARS in multiorgan failure secondary to respiratory illnesses and the possible influence on inflammatory mediators and cytokines. The application of 4 intermittent MARS treatments (8 h each, mean blood flow rate 180 ml/min) on 4 consecutive days resulted in an immediate improvement of clinical conditions within the treatment days. The further improvement of organ functions allowed withdrawing the patient from ventilatory support 13 days after start of MARS, and 44 days after admission he was discharged home with completely resolved organ functions and laboratory abnormalities. SARS is a severe form of the epidemic outbreak of atypical pneumonia which remains poorly defined regarding etiology and special therapy recommendations. However, the development and aggravation of this ARDS-like severe acute respiratory syndrome is pathologically associated with the systemic inflammatory response syndrome (SIRS) which may then mediate or cause MODS. To our knowledge, this is the first report of an application of MARS therapy in MODS which was probably induced by SARS in a patient in China which improved the clinical condition of the patient in multi-organ failure secondary to respiratory failure indicating that MARS might be an additional therapeutic option in multiorgan failure induced by SARS.


Growing popularity of gainsharing brings pushback from manufacturers.


title: Purification and molecular weight determination of measles virus genomic RNA.

text: A purification procedure for genomic measles virus RNA, free of contaminating smaller RNA and of DNA, is described. Viral nucleocapsids were prepared from MA160 cells infected in spinner cultures with measles virus (Edmonston strain). Nucleic acid was extracted, treated with DNase and RNA sedimenting at about 50S in sucrose gradients was isolated. This method yielded 0.5 to 1.5 micrograms of genomic RNA per litre of culture. A molecular weight of 4.5 X 10(6) was determined by gel electrophoresis under fully denaturing conditions.


title: Prevalence and mutation analysis of the spike protein in feline enteric coronavirus and feline infectious peritonitis detected in household and shelter cats in western Canada.

text: Feline infectious peritonitis (FIP) is a fatal disease for which no simple antemortem diagnostic assay is available. A new polymerase chain reaction (PCR) test has recently been developed that targets the spike protein region of the FIP virus (FIPV) and can identify specific mutations (M1030L or S1032A), the presence of which indicates a shift from feline enteric coronavirus (FeCV) to FIPV. This test will only be useful in the geographical region of interest, however, if the FIP viruses contain these mutations. The primary objective of this study was to determine the presence of the M1030L or S1032A mutations in FeCV derived from stool samples from a selected group of healthy cats from households and shelters and determine how many of these cats excrete FeCV. The secondary objective was to evaluate how often these specific FIPV mutations were present in tissue samples derived from cats diagnosed with FIP at postmortem examination. Feline enteric coronavirus (FeCV) was detected in 46% of fecal samples (86/185), all were FeCV type 1, with no difference between household or shelter cats. Only 45% of the FIPV analyzed contained the previously reported M1030L or S1032A mutations. It should be noted that, as the pathological tissue samples were opportunistically obtained and not specifically obtained for PCR testing, caution is warranted in interpreting these data.


title: ANZICS guiding principles for complex decision making during the COVID-19 pandemic.

text: The global 2019 coronavirus disease (COVID-19) pandemic has led to major challenges in clinical decision making when the demand for intensive care exceeds local capacity. In order to promote consistent, transparent, objective and ethical decision making, the Australian and New Zealand Intensive Care Society (ANZICS) formed a committee to urgently develop guidelines outlining key principles that should be utilised during the pandemic. This guidance is intended to support the practice of intensive care specialists during the COVID-19 pandemic and to promote the development of local admission policies that should be endorsed by health care organisations and relevant local authorities.


title: Reviewing lessons learnt of SARS in Singapore during planning for influenza pandemic.

text: There were many lessons learnt in Singapore's fight against SARS, and they have proven to be all the more important in our preparations for an influenza pandemic. The following lessons are discussed in this paper including: the widespread ramifications among the various sectors in Singapore (individuals, organizations, community and economy), the first principles of outbreak response, need for enhanced infectious disease control measures, high demands on the healthcare system, the role of management policies implementation and dissemination, multidisciplinary involvement, importance of communication, and business continuity planning.


title: Interference of coronavirus infection by expression of immunoglobulin G (IgG) or IgA virus-neutralizing antibodies.

text: Immunoglobulin gene fragments encoding the variable modules of the heavy and light chains of a transmissible gastroenteritis coronavirus (TGEV)-neutralizing monoclonal antibody (MAb) have been cloned and sequenced. The selected MAb recognizes a highly conserved viral epitope and does not lead to the selection of neutralization escape mutants. The sequences of MAb 6A.C3 kappa and gamma 1 modules were identified as subgroup V and subgroup IIIC, respectively. The chimeric immunoglobulin genes encoding the variable modules from the murine MAb and constant modules of human gamma 1 and kappa chains were constructed by reverse transcriptase PCR. Chimeric immunoglobulins were stably or transiently expressed in murine myelomas or COS cells, respectively. The secreted recombinant antibodies had radioimmunoassay titers (i.e., the highest dilution giving a threefold increase over the background) higher than 10(3) and reduced the infectious virus more than 10(4)-fold. Recombinant dimeric immunoglobulin A (IgA) showed a 50-fold enhanced neutralization of TGEV relative to a recombinant monomeric IgG1 which contained the identical antigen binding site. Stably transformed epithelial cell lines which expressed either recombinant IgG or IgA TGEV-neutralizing antibodies reduced virus production by > 10(5)-fold after infection with homologous virus, although a residual level of virus production (< 10(2) PFU/ml) remained in less than 0.1% of the cells. This low-level persistent infection was shown not to be due to the selection of neutralization escape mutants. The implications of these findings for somatic gene therapy with recombinant antibodies are discussed.


title: Interaction of a peptide from the pre-transmembrane domain of the severe acute respiratory syndrome coronavirus spike protein with phospholipid membranes.

text: The severe acute respiratory syndrome coronavirus (SARS-CoV) envelope spike (S) glycoprotein, a Class I viral fusion protein, is responsible for the fusion between the membranes of the virus and the target cell. In order to gain new insight into the protein membrane alteration leading to the viral fusion mechanism, a peptide pertaining to the putative pre-transmembrane domain (PTM) of the S glycoprotein has been studied by infrared and fluorescence spectroscopies regarding its structure, its ability to induce membrane leakage, aggregation, and fusion, as well as its affinity toward specific phospholipids. We demonstrate that the SARS-CoV PTM peptide binds to and interacts with phospholipid model membranes, and, at the same time, it adopts different conformations when bound to membranes of different compositions. As it has been already suggested for other viral fusion proteins such as HIV gp41, the region of the SARS-CoV protein where the PTM peptide resides could be involved in the merging of the viral and target cell membranes working synergistically with other membrane-active regions of the SARS-CoV S glycoprotein to heighten the fusion process and therefore might be essential for the assistance and enhancement of the viral and cell fusion process.


title: [Theory and practice of aging upon COVID-19 pandemic.]

text: Never before in history, aging was such a significant factor for epidemics as it is now for the current COVID-19 pandemic, which features a drastic shift of mortality towards older ages. Our analysis of data on COVID-19-related mortality in Spain, Italy, and Sweden has shown that, in the range of 30 to 90 years of age, each dependency of the logarithm of mortality upon age is linear, and all regression lines are strictly parallel to those related to the total mortality in accordance with the Gompertz law. In all cases, irrespective of the stage and place of epidemic, mortality doubling time in this age range is close to 7,5 years. The rates of being infected with the SARS-CoV-2 coronavirus and of being diagnosed due to the symptomatic manifestations of the infection are dependent on age to a far lesser degree. With account for these observations, three messages are put forth: 1) Older persons are the principal victims of both SARS-CoV-2 and measures undertaken to control its spread; 2) Older persons are not the principal driving force of SARS-CoV-2 spread; 3) Older persons can and should be engaged in combating the pandemic and its consequences; however, not via selective social distancing and other discriminative measures. People aged over 65 years constitute a significant part of the current population. They have specific interests and needs, which deserve no less respect than those of any other age group. This includes the right for the quality of life that remains sustained under the emergency conditions. Since the prospects for controlling the SARS-CoV-2 are dubious, those in charge of decisions concerning «people aged above 65» should mind that currently, unlike in the medieval ages, 65+ is the individual future of almost everyone.


title: Severe acute respiratory syndrome: responses of the healthcare system to a global epidemic.

text: PURPOSE OF REVIEW To evaluate recent developments in the response of the healthcare system to the threat of severe acute respiratory syndrome in the context of the Toronto experience. RECENT FINDINGS The severe acute respiratory syndrome outbreak affected over 8000 people killing 774 by July 2003. It particularly affected those providing health care. The initial response of the healthcare system was to use infection control techniques to prevent transmission to staff and patients. This had a dramatic impact on healthcare provision for non-severe acute respiratory syndrome patients during this period. SUMMARY The ongoing response to severe acute respiratory syndrome is continued effective infection control education, development of local and global policies for reacting to a severe acute respiratory syndrome-like outbreak in the future, and continued work on a vaccine for severe acute respiratory syndrome.


Evolving Techniques for Bariatric Surgery and Endoscopy.


title: COVID-19 Special Column: Principles Behind the Technology for Detecting SARS-CoV-2, the Cause of COVID-19.

text: Nationwide shortages of tests that detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and diagnose coronavirus disease 2019 (COVID-19) have led the US Food and Drug Administration (FDA) to significantly relax regulations regarding COVID-19 diagnostic testing. To date the FDA has given emergency use authorization (EUA) to 48 COVID-19 in vitro diagnostic tests and 21 high complexity molecular-based laboratory developed tests, as well as implemented policies that give broad authority to clinical laboratories and commercial manufacturers in the development, distribution, and use of COVID-19 diagnostic tests. Currently, there are 2 types of diagnostic tests available for the detection of SARS-CoV-2: (1) molecular and (2) serological tests. Molecular detection of nucleic acid (RNA or DNA) sequences relating to the suspected pathogen is indicative of an active infection with the suspected pathogen. Serological tests detect antibodies against the suspected pathogen, which are produced by an individual's immune system. A positive serological test result indicates recent exposure to the suspected pathogen but cannot be used to determine if the individual is actively infected with the pathogen or immune to reinfection. In this article, the SARS-CoV-2 diagnostic tests currently approved by the FDA under EUA are reviewed, and other diagnostic tests that researchers are developing to detect SARS-CoV-2 infection are discussed.


title: [Analysis and causation discussion of 185 severe acute respiratory syndrome dead cases].

text: OBJECTIVE To proceed the retrospective analysis of the dead reasons of patients with severe acute respiratory syndrome (SARS), in order to provide experience for decreasing mortality of SARS patients in clinic. METHODS Retrospective analyze of 185 dead cases of SARS patients, including age, basic diseases, laboratory tests, radiological test and therapy. RESULTS There were 102 men and 83 women in 185 dead cases of SARS, in which average age was 56.4 years, 85 cases were older than 60 years. 147 cases (79.5%) had basic diseases, cardiovascular disease (57 cases) was at the first Place; diabetes mellitus (41 cases) was at the second Place; cerebrovascular disease (19 cases) was at the third place; 15 patients had alimentary disease. 52 patients (28.1%) suffered from complications, which were acute respiratory distress syndrome (ARDS), secondary infection and multiple organ dysfunctional syndrome (MODS) by order. Some of the patients having complete information showed that life symptoms were stabilization. Hemogram and blood gas analysis showed that blood sugar and white blood cell (WBC) increased (P<0.001); lymphocyte decreased significantly (P<0.005) and PaO(2), PaO(2)/FiO(2) decreased Progressively (P<0.005 P<0.001) in late stage of SARS versus just entering the hospital. Half patients showed the abnormality of biochemical indexes. CONCLUSION Age and basic diseases are sensitive indexes for predicting dead tendency of SARS. Lung injury, obstinate hypoxemia, abnormality metabolize of hyperglycemia and misusing glucocorticoids and antibiotic leading to secondary infection are the main reasons for SARS patients dying.


title: Reverse quality management: developing evidence-based best practices in health emergency management.

text: The British Columbia Ministry of Health's Framework for Core Functions in Public Health was the catalyst that inspired this review of best practices in health emergency management. The fieldwork was conducted in the fall of 2005 between hurricane Katrina and the South Asia earthquake. These tragedies, shown on 24/7 television news channels, provided an eyewitness account of disaster management, or lack of it, in our global village world. It is not enough to just have best practices in place. There has to be a governance structure that can be held accountable. This review of best practices lists actions in support of an emergency preparedness culture at the management, executive, and corporate/governance levels of the organization. The methodology adopted a future quality management approach of the emergency management process to identify the corresponding performance indictors that correlated with practices or sets of practices. Identifying best practice performance indictors needed to conduct a future quality management audit is described as reverse quality management. Best practices cannot be assessed as stand-alone criteria; they are influenced by organizational culture. The defining of best practices was influenced by doubt about defining a practice it is hoped will never be performed, medical staff involvement, leadership, and an appreciation of the resources required and how they need to be managed. Best practice benchmarks are seen as being related more to "measures" of performance defined locally and agreed on by 2 or more parties rather than to achieving industrial standards. Relating practices to performance indicators and then to benchmarks resulted in the development of a Health Emergency Management Best Practices Matrix that lists specific practice in the different phases of emergency management.


title: Severe acute respiratory syndrome (SARS): a review.

text: Severe acute respiratory syndrome (SARS) is a newly emerged disease that rapidly spread around the world. The disease originated in southern China and a novel coronavirus (SARS CoV) has been implicated as the causative organism. The path this virus took to set up human infection remains a mystery, though preliminary data point to origins in an animal reservoir. Nosocomial transmission of SARS CoV has been a striking feature in this epidemic. The clinical illness is similar to many acute respiratory infections, although a large proportion of patients show a rapid deterioration with respiratory distress towards the end of the second week of illness. The management principles are broadly similar to treating any community acquired pneumonia but the infection control measures take a pivotal role. There is no proven antiviral agent against SARS CoV. The most remarkable feature about the SARS epidemic was the speed with which the global community acted in a coordinated way to control it.


title: Important roles for gamma interferon and NKG2D in gammadelta T-cell-induced demyelination in T-cell receptor beta-deficient mice infected with a coronavirus.

text: gammadelta T cells mediate demyelination in athymic (nude) mice infected with the neurotropic coronavirus mouse hepatitis virus strain JHM. Now, we show that these cells also mediate the same process in mice lacking alphabeta T cells (T-cell receptor beta-deficient [TCRbeta(-/-)] mice) and demyelination is gamma interferon (IFN-gamma) dependent. Most strikingly, our results also show a major role for NKG2D, expressed on gammadelta T cells, in the demyelinating process with in vivo blockade of NKG2D interactions resulting in a 60% reduction in demyelination. NKG2D may serve as a primary recognition receptor or as a costimulatory molecule. We show that NKG2D(+) gammadelta T cells in the JHM-infected central nervous system express the adaptor molecule DAP12 and an NKG2D isoform (NKG2D short), both required for NKG2D to serve as a primary receptor. These results are consistent with models in which gammadelta T cells mediate demyelination using the same effector cytokine, IFN-gamma, as CD8 T cells and do so without a requirement for signaling through the TCR.


Health is a global issue.


title: Implementing the general use of dissection devices in thyroid surgery from prospective randomized trial to daily use.

text: Routine use of vascular sealing and dissecting devices was implemented in our Tertiary Center of Endocrine Surgery with a high volume of thyroid operations. Beginning with a prospective randomized trial on 82 patients that compared thyroid surgery with assistance of a new, high-frequency-powered electrothermal device (Precise™, Covidien, Boulder, Colorado) to conventional thyroid surgery, approximately 30% of operation time was saved in the group using the device. Surgeons were then free to decide whether to use LigaSure (Precise™), an ultrasonic device (Focus™, Ethicon Endo Surgery, Cincinnati, Ohio), or the classic tie-and-knotting procedure. This change of procedure resulted in an increase of surgery with devices, when compared to classic procedures from 20.2% to 98.4%, during a period of 5 years with 2,591 patients. The equal results of device-supported thyroid surgery were demonstrated when 100 consecutive patients with Precise™ and Focus™ were compared, by measuring time of surgery (-36% or 43 min) and postoperative morbidity. The authors advocate the use of vascular sealing and dissecting devices for thyroid operations and, especially, for surgical units with high operative frequency.


title: Local recurrence and complications after percutaneous radiofrequency ablation of hepatocellular carcinoma: a retrospective cohort study focused on tumor location.

text: We conducted a retrospective cohort study to investigate the predisposing factors for local recurrence and complications after percutaneous radiofrequency ablation (RFA) of hepatocellular carcinoma (HCC). HCC patients (n＝397) consecutively treated with RFA (256 males, 141 females, median age 69 years) were enrolled. In these patients, 1,455 nodules (median size 17mm) were ablated. Predisposing factors for overall recurrence and local recurrence in the context of tumor location and complications were examined. Local recurrence was observed for 113 of the 1,455 nodules. The 1-, 3- and 5-year local recurrence rates were 2.2%, 7.4% and 9.5%, respectively. A multivariate Cox proportional hazard analysis revealed that large tumor size (＞2cm), tumor location (adjacent to the major portal branch or hepatic vein), and small ablated margin (＜3mm) were independent predisposing factors for local recurrence after RFA (HR＝1.70-2.81). Tumor location (adjacent to the major portal branch, hepatic vein, or diaphragm) was also revealed as a risk factor for liver damage due to RFA. HCC adjacent to the major portal vein or hepatic vein was associated with a higher risk for local recurrence and for complications;therefore, special precautions are necessary when applying RFA to HCC near vessels even when the tumors are located at an easy-to-puncture site.


title: Does social capital matter when medical professionals encounter the SARS crisis in a hospital setting.

text: This study examined whether social capital can enhance an individual's ability in reducing emotional exhaustion and job tension when medical professionals encounter a crisis such as Severe Acute Respiratory Syndrome (SARS).


title: The role of wildlife in emerging and re-emerging zoonoses.

text: There are huge numbers of wild animals distributed throughout the world and the diversity of wildlife species is immense. Each landscape and habitat has a kaleidoscope of niches supporting an enormous variety of vertebrate and invertebrate species, and each species or taxon supports an even more impressive array of macro- and micro-parasites. Infectious pathogens that originate in wild animals have become increasingly important throughout the world in recent decades, as they have had substantial impacts on human health, agricultural production, wildlife-based economies and wildlife conservation. The emergence of these pathogens as significant health issues is associated with a range of causal factors, most of them linked to the sharp and exponential rise of global human activity. Among these causal factors are the burgeoning human population, the increased frequency and speed of local and international travel, the increase in human-assisted movement of animals and animal products, changing agricultural practices that favour the transfer of pathogens between wild and domestic animals, and a range of environmental changes that alter the distribution of wild hosts and vectors and thus facilitate the transmission of infectious agents. Two different patterns of transmission of pathogens from wild animals to humans are evident among these emerging zoonotic diseases. In one pattern, actual transmission of the pathogen to humans is a rare event but, once it has occurred, human-to-human transmission maintains the infection for some period of time or permanently. Some examples of pathogens with this pattern of transmission are human immunodeficiency virus/acquired immune deficiency syndrome, influenza A, Ebola virus and severe acute respiratory syndrome. In the second pattern, direct or vector-mediated animal-to-human transmission is the usual source of human infection. Wild animal populations are the principal reservoirs of the pathogen and human-to-human disease transmission is rare. Examples of pathogens with this pattern of transmission include rabies and other lyssaviruses, Nipah virus, West Nile virus, Hantavirus, and the agents of Lyme borreliosis, plague, tularemia, leptospirosis and ehrlichiosis. These zoonotic diseases from wild animal sources all have trends that are rising sharply upwards. In this paper, the authors discuss the causal factors associated with the emergence or re-emergence of these zoonoses, and highlight a selection to provide a composite view of their range, variety and origins. However, most of these diseases are covered in more detail in dedicated papers elsewhere in this Review.


title: Estimating in real time the efficacy of measures to control emerging communicable diseases.

text: Controlling an emerging communicable disease requires prompt adoption of measures such as quarantine. Assessment of the efficacy of these measures must be rapid as well. In this paper, the authors present a framework to monitor the efficacy of control measures in real time. Bayesian estimation of the reproduction number R (mean number of cases generated by a single infectious person) during an outbreak allows them to judge rapidly whether the epidemic is under control (R < 1). Only counts and time of onset of symptoms, plus tracing information from a subset of cases, are required. Markov chain Monte Carlo and Monte Carlo sampling are used to infer the temporal pattern of R up to the last observation. The operating characteristics of the method are investigated in a simulation study of severe acute respiratory syndrome-like outbreaks. In this particular setting, control measures lacking efficacy (R > or = 1.1) could be detected after 2 weeks in at least 70% of the epidemics, with less than a 5% probability of a wrong conclusion. When control measures are efficacious (R = 0.5), this situation may be evidenced in 68% of the epidemics after 2 weeks and 92% of the epidemics after 3 weeks, with less than a 5% probability of a wrong conclusion.


title: A case of severe monkeypox virus disease in an American child: emerging infections and changing professional values.

text: Monkeypox virus disease is a rare zoonosis that until recently was limited to Central Africa. We describe the clinical features of the third child in the United States reported with this newly emerging infection. This child was part of a large cluster of individuals in the Midwest infected by prairie dogs that had contracted the virus when housed with infected small mammals imported from Africa. The differential and laboratory diagnoses and the difficulty finding physicians and nurses to care for this patient are discussed.


title: Is phenomenology the best approach to health research?

text: Much research at the National Institutes of Health--for example, the NIH Roadmap (http://nihroadmap.nih.gov/)--has focused on aspects of systems biology and the application of new technologies, which might best be considered descriptive or phenomenological. Unfortunately, etiologic research and social, behavioral, and environmental population studies are at risk of becoming second-class research. In particular, the Roadmap does not address the need for studies of unique populations, determinants of the large variations in disease among populations and over time, and the long incubation period for many diseases. Success in reducing disease in the population will depend on linking the enormous potential of phenomenological methods to excellent etiologic and social/behavioral studies. The phenomenological approach alone will improve our descriptions of disease but may not result in reducing disease burden in human populations.


Identification of a cellular receptor for fish rhabdoviruses.


title: Development of an immunochromatographic strip for serological diagnosis of Porcine hemagglutinating encephalomyelitis virus.

text: An immunochromatographic strip was developed for the detection of an antibody against Porcine hemagglutinating encephalomyelitis virus (PHEV). Colloidal gold-labeled rabbit anti-pig immunoglobulin G (IgG) was used as the detection reagent, and the PHEV recombinant antigens and goat anti-rabbit IgG were coated on the prototype strip and the control lines, respectively. The immunochromatographic strip was capable of specifically detecting PHEV antibodies in serum with a hemagglutination inhibition (HI) titer of 2 within 10 min. Storage of the strips at room temperature for 6 months or at 4°C for 12 months did not change their sensitivity and specificity. Using HI as a reference test, the relative specificity and sensitivity of the immunochromatographic strip were determined to be 93.41% and 98.42%, respectively. There was a strong agreement between the results obtained by HI and the immunochromatographic strips (κ = 0.926). Additionally, there was a strong agreement between enzyme-linked immunosorbent assay and immunochromatographic strips (κ = 0.929). When the immunochromatographic strip was used for serological diagnosis of 1,117 serum samples in Jilin Province in China, the seropositivity ranged from 6.5% in the Liaoyuan District to 81.6% in the Changchun District. Furthermore, many piglets were seropositive to PHEV, indicating the possible transfer of maternal antibodies via the colostrum. Based on the high specificity, sensitivity, and stability of the immunochromatographic strip, it would be suitable for on-site detection of PHEV antibodies in order to monitor PHEV infections in an animal population.


title: Dissection study on the severe acute respiratory syndrome 3C-like protease reveals the critical role of the extra domain in dimerization of the enzyme: defining the extra domain as a new target for design of highly specific protease inhibitors.

text: The severe acute respiratory syndrome (SARS) 3C-like protease consists of two distinct folds, namely the N-terminal chymotrypsin fold containing the domains I and II hosting the complete catalytic machinery and the C-terminal extra helical domain III unique for the coronavirus 3CL proteases. Previously the functional role of this extra domain has been completely unknown, and it was believed that the coronavirus 3CL proteases share the same enzymatic mechanism with picornavirus 3C proteases, which contain the chymotrypsin fold but have no extra domain. To understand the functional role of the extra domain and to characterize the enzyme-substrate interactions by use of the dynamic light scattering, circular dichroism, and NMR spectroscopy, we 1) dissected the full-length SARS 3CL protease into two distinct folds and subsequently investigated their structural and dimerization properties and 2) studied the structural and binding interactions of three substrate peptides with the entire enzyme and its two dissected folds. The results lead to several findings; 1) although two dissected parts folded into the native-like structures, the chymotrypsin fold only had weak activity as compared with the entire enzyme, and 2) although the chymotrypsin fold remained a monomer within a wide range of protein concentrations, the extra domain existed as a stable dimer even at a very low concentration. This observation strongly indicates that the extra domain contributes to the dimerization of the SARS 3CL protease, thus, switching the enzyme from the inactive form (monomer) to the active form (dimer). This discovery not only separates the coronavirus 3CL protease from the picornavirus 3C protease in terms of the enzymatic mechanism but also defines the dimerization interface on the extra helical domain as a new target for design of the specific protease inhibitors. Furthermore, the determination of the preferred solution conformation of the substrate peptide S1 together with the NMR differential line-broadening and transferred nuclear Overhauser enhancement study allows us to pinpoint the bound structure of the S1 peptide.


title: Aviation Medicine: global historical perspectives and the development of Aviation Medicine alongside the growth of Singapore's aviation landscape.

text: Aviation Medicine traces its roots to high altitude physiology more than 400 years ago. Since then, great strides have been made in this medical specialty, initially catalysed by the need to reduce pilot medical attrition during the World Wars, and more recently, fuelled by the explosive growth in globalised commercial air travel. This paper traces the historical milestones in Aviation Medicine, and maps its development in Singapore since the 1960s. Advancements in military aviation platforms and technology as well as the establishment of Singapore as an international aviation hub have propelled Aviation Medicine in Singapore to the forefront of many domains. These span Aviation Physiology training, selection medical standards, performance maximisation, as well as crew and passenger protection against communicable diseases arising from air travel. The year 2011 marks the centennial milestone of the first manned flight in Singapore, paving the way for further growth of Aviation Medicine as a mature specialty in Singapore.


title: Internal entry of ribosomes on a tricistronic mRNA encoded by infectious bronchitis virus.

text: mRNA3 specified by the coronavirus infectious bronchitis virus appears to be functionally tricistronic, having the capacity to encode three small proteins (3a, 3b, and 3c) from separate open reading frames (ORFs). The mechanism by which this can occur was investigated through in vitro translation studies using synthetic mRNAs containing the 3a, 3b, and 3c ORFs, and the results suggest that translation of the most distal of the three ORFs, that for 3c, is mediated by an unconventional, cap-independent mechanism involving internal initiation. This conclusion is based on several observations. A synthetic mRNA whose peculiar 5' end structure prevents translation of the 5'-proximal ORFs (3a and 3b) directs the synthesis of 3c normally. Translation of 3c, unlike that of 3a and 3b, was insensitive to the presence of the 5' cap analog 7-methyl-GTP, and it was unaffected by alteration of the sequence contexts for initiation on the 3a and 3b ORFs. Finally, an mRNA in which the 3a/b/c infectious bronchitis virus coding region was placed downstream of the influenza A virus nucleocapsid protein gene directed the efficient synthesis of 3c as well as nucleocapsid protein, whereas initiation at 3a and 3b could not be detected. Expression of the 3c ORF from this mRNA, however, was abolished when the 3a and 3b coding region was deleted, indicating that 3c initiation is dependent on upstream sequence elements which together may serve as a ribosomal internal entry site similar to those described for picornaviruses.


March/April 2020: Special COVID-19 Issue.


The novel human coronaviruses NL63 and HKU1.


title: Isolation and chemiluminescent properties of ferret (Mustela putorius furo) polymorphonuclear cells.

text: Ferret polymorphonuclear cells (PMNs) and peripheral blood mononuclear cells (PBMCs) were separated from whole blood by density gradient centrifugation. Using a 50% Percoll solution (density=1.066), PMNs and PBMCs were successfully isolated after centrifugation; the purities of the PMNs and PBMCs were 94.2% and 95.6%, respectively. To evaluate the function of isolated ferret PMNs, we measured the superoxide generation with a MCLA-dependent chemiluminescence assay. The isolated ferret PMNs responded to phorbol 12-myristate 13-acetate (PMA) with kinetics similar to that of human PMNs. The ferret PMNs did not respond to N-formyl-Met-Leu-Phe (fMLF), unlike human PMNs, which rapidly responded. Thus, authors established a method for the rapid separation of highly purified populations of functional PMNs from the whole blood of ferrets.


title: [Emergence of transmissible disorders, a continuous process--a new type of viral meningoencephalitis].

text: In the interval 1994-1999, in Australia, Malaysia and Singapore, epizootic and epidemiological episodes of meningoencephalitis and severe acute respiratory syndromes were reported. Highly lethal in horses, swine and humans, the episodes were proved to be caused by the "new" viruses Hendra (HeV) and Nipah (NiV). At the same time three "new" viral agents have been isolated: Lyssavirus, Menanglevirus and Tupaia paramyxovirus. The intense contemporary circulation of people, animals and food products together with changes in human ecosystem favor new relations between humans and the "natural reservoirs" of biologic agents with a pathogenic potential for domestic and peridomestic animals and humans.


title: The threat of severe acute respiratory syndrome (SARS).

text: Severe acute respiratory syndrome (SARS) is a recently recognized infectious disease associated with severe morbidity and mortality. It presents with non-specific signs and symptoms and because no definitive laboratory test is readily available, it poses a great risk to healthcare workers as well as difficulty in quarantine. The global response has been coordinated and enthusiastic in trying to understand and control this disease. Severe acute respiratory syndrome poses a threat to the Caribbean because of easy and convenient travel and the vibrant tourist industry.


title: Acute respiratory distress syndrome.

text: BACKGROUND Acute respiratory distress syndrome (ARDS) is a common diagnosis among children admitted to pediatric intensive care units. This heterogeneous disorder has numerous pulmonary and non-pulmonary causes and is associated with a significant risk of mortality. Many supportive therapies exist for ARDS. SEARCH: Literature search was performed by using the key words ARDS and related topics on the Pubmed search engine maintained by the National Heart, Lung, Blood Institute. Pediatric randomized controlled trials that have been published in the last 10 years were included. Emphasis was placed on pediatric literature, although sentinel adult studies have been included. Most of the evidence presented is of levels I and II. RESULTS Low tidal volume is the only strategy that has consistently improved outcome in ARDS. A tidal volume of ≤ 6 mL/kg predicted body weight should be used. Ventilator induced lung injury may result in systemic effects with multi-system organ failure, and all efforts should be made to minimize this. Positive end-expiratory pressure should be used to judiciously maintain lung recruitment. There is insufficient evidence to routinely use high frequency ventilation, prone positioning, or inhaled nitric oxide. Calfactant therapy is promising and may be considered in children with direct lung injury and ARDS. Current literature does not support routine use of corticosteroids for non-resolving ARDS.


title: Laboratory procedures for the diagnosis of gastrointestinal tract diseases of dogs and cats.

text: An increasing number of laboratory tests are available for diagnosis of gastrointestinal tract diseases in dogs and cats. Use of these tests can lead to more accurate and rapid diagnoses. This review discusses laboratory tests, both new and old, and the role they currently play in the evaluation of animals presented with gastrointestinal problems. A minimum database helps assess the severity of the disorder, detect extra-gastrointestinal causes of problems and assists in formulating diagnostic and therapeutic plans. Faecal examination remains one of the most important diagnostic procedures in the investigation of gastrointestinal problems. Zinc sulphate faecal flotation is an excellent routine screening technique for helminth and protozoal infections, including giardiasis. Rectal cytology can assist in the diagnosis of large bowel disorders. Interpretation of faecal immunodiagnostic tests is hampered by insufficient knowledge of test sensitivities and specificities. Routine faecal cultures are not warranted and faecal occult blood tests are rarely indicated. Serum tests for gastric inflammation are now under development. The serum trypsin-like immunoreactivity test remains the gold standard technique for the diagnosis of exocrine pancreatic insufficiency. Breath hydrogen tests can be helpful in assessing the functional relevance of mild abnormalities in small-bowel biopsy specimens. Subnormal concentrations of serum cobalamin appear to be more specific indicators of gastrointestinal disease in cats than in dogs. Tests for small intestinal bacterial overgrowth remain controversial and assessment of gastrointestinal permeability has yet to prove its value in the diagnostic assessment of companion animals with gastrointestinal problems. Faecal alpha1-protease inhibitor shows promise for the diagnosis of protein-losing enteropathy.


title: Detecting robust patterns in the spread of epidemics: a case study of influenza in the United States and France.

text: In this paper, the authors develop a method of detecting correlations between epidemic patterns in different regions that are due to human movement and introduce a null model in which the travel-induced correlations are cancelled. They apply this method to the well-documented cases of seasonal influenza outbreaks in the United States and France. In the United States (using data for 1972-2002), the authors observed strong short-range correlations between several states and their immediate neighbors, as well as robust long-range spreading patterns resulting from large domestic air-traffic flows. The stability of these results over time allowed the authors to draw conclusions about the possible impact of travel restrictions on epidemic spread. The authors also applied this method to the case of France (1984-2004) and found that on the regional scale, there was no transportation mode that clearly dominated disease spread. The simplicity and robustness of this method suggest that it could be a useful tool for detecting transmission channels in the spread of epidemics.


title: Taste and smell as chemosensory dysfunctions in COVID-19 infection.

text: PURPOSE To review the literature on the presence of two clinical manifestations in patients presenting COVID-19 (SARS-CoV-2) infection: loss of taste (ageusia) and loss of smell (anosmia). METHODS PubMed and EMBASE were searched and studies were selected starting from November, 2019 until April 2020; also, the references of the selected articles were evaluated for methodological quality. RESULTS Of the 19 studies analyzed, five were included to evaluate the presence of ageusia and/or anosmia as symptoms in patients who were tested and resulted positive for the SARS-CoV-2 virus. In a total of 10,818 patients, 8,823 presented ageusia (81.6%; range 5.6%-88%) and 8,088 presented anosmia (74.8%; range 5.1-85.6%). Only one study recorded both symptoms with a percentage of 18.6%. CLINICAL SIGNIFICANCE This systematic review demonstrated significant presence of ageusia and anosmia in the patients with COVID-19 infection. These symptoms may be considered as the first manifestation of the infection.


title: Coronavirus as an agent of neonatal calf diarrhea in a Chinese dairy cattle farm.

text: Neonatal calf diarrhea is a significant health problem in dairy industry in the whole world. Although the aetiological agents of neonatal diarrhea are diverse, it is obvious that viruses play an important role. One of them is bovine coronavirus (BCV). Since BCV was discovered (MEBUS et al., 1969), there have been many published reports describing the virus and its importance in calves. In China, we first investigated the seroepidemiological status of BCV infections in cattle (YAO et al., 1990). The results showed that the incidence of infections with BCV in cattle in different chinese regions are high. Further work on direct diagnosis of BCV as a causative agent of diarrhea was therefore necessary. In the last years we have developed diagnostic methods including a monoclonal antibody-based ELISA to detect BCV and other diarrheal viruses. In this study, the diagnosis of an outbreak of BCV in chinese dairy cattle is described.


Apolipoprotein D: an overview of its role in aging and age-related diseases.


title: Self-assembly of the recombinant capsid protein of a bovine norovirus (BoNV) into virus-like particles and evaluation of cross-reactivity of BoNV with human noroviruses.

text: None of the enteric caliciviruses except Po/Sapo/GIII/Cowden/80/US replicates in cell culture, which complicates efforts to develop control strategies or to study viral replication. To develop serological assays for bovine noroviruses (BoNVs) and to determine the cross-reactivity of BoNV with human noroviruses, we generated two recombinant baculoviruses, rCV186-OH and rJNCV, to express the capsid genes of Bo/CV186-OH/00/US (Norovirus genogroup III [GIII], genotype 2 [GIII/2]). rCV186-OH expressed the expected 57-kDa capsid protein, but rJNCV expressed a truncated capsid protein of 35 kDa. Sequence analysis of rJNCV identified a single nucleotide deletion in the P domain of the capsid gene, which introduced a stop codon at amino acid 323. The recombinant capsid protein produced by rCV186-OH but not that produced by rJNCV self-assembled into virus-like particles (VLPs) similar to native BoNV. An antibody-capture enzyme-linked immunosorbent assay (ELISA) and antigen-capture ELISA (Ag-ELISA) detected serum antibody and antigen, respectively, from calves infected with Bo/CV186-OH/00/US but not antibodies or antigens to other enteric viruses. In other tests of the GIII/2 BoNV Ag-ELISA, no cross-reactivity was observed with VLPs from one GI and four GII human noroviruses and porcine sapovirus Cowden strain. Because, like human noroviruses, BoNVs do not grow in cell culture, the BoNV VLPs will be useful in the serological assays described for the detection of BoNV antibody and antigen. Consistent with the phylogenetic analysis of the capsid genes of bovine and human noroviruses (M. G. Han, J. R. Smiley, C. Thomas, and L. J. Saif, J. Clin. Microbiol. 42:5214-5224, 2004), the results suggest that GIII/2 BoNV does not share significant antigenic relationships with the five characterized human noroviruses tested.


title: Polycation-mediated delivery of siRNAs for prophylaxis and treatment of influenza virus infection.

text: Influenza A virus causes one of the most prevalent infections in humans. In a typical year, 10-20% of the population of the US is infected by influenza virus, resulting in up to 40,000 deaths and 200,000 hospitalisations. Vaccination is the most effective preventative measure that can protect 70-90% of healthy adults aged < 65; however, the protection rate is much lower in those most susceptible to infection, namely infants, the elderly and individuals with weakened immune systems. Although four drugs have been approved by the FDA for use as prophylaxis and/or treatment of influenza, concerns about their side effects and the emergence of drug-resistant viruses persist. RNA interference (RNAi), an emerging method of post transcriptional gene silencing, appears ideal for the prevention and treatment of influenza. RNAi in mammals can be mediated by short interfering RNAs (siRNAs) of approximately 21-27 nucleotides in length. The authors have previously shown that siRNAs specific for conserved regions of the influenza virus genome are potent inhibitors of influenza virus replication in both cell lines and chicken embryos. This review discusses the recent progress in the in vivo inhibition of influenza virus by the delivery of siRNAs mediated by non-viral vectors, and the prospects of this strategy for prophylaxis and treatment of influenza infection in humans.


Zika virus infection: global update on epidemiology and potentially associated clinical manifestations.


title: Asymptomatic stage of human immunodeficiency virus infection is the optimal timing for its management with Traditional Chinese Medicine.

text: Acquired immune deficiency syndrome (AIDS) is a worldwide public health problem and an incurable disease. The roles of Traditional Chinese Medicine (TCM) on AIDS had been recognized and accepted by more and more people. Being a relatively long period, to initiate combination antiretroviral therapy (cART) for the asymptomatic human immunodeficiency virus (HIV) infection patient is not consensus. According to TCM theory, not only the imbalance of Yin and Yang but also the disharmony of internal body function and external natural environment had occurred in the body of the patient, the imbalance and the disharmony should be adjusted by the means of the treatment based on the patters identified among the symptoms in terms of TCM. We aimed to analyze the feature of patients with asymptomatic infection, and compare the advantages and disadvantages of the us- age of cART for asymptomatic infection, explored the possible mechanism underlying TCM treatment of the asymptomatic infection, by reviewing the TCM treatment progress on asymptomatic HIV infection.


title: The spike protein of murine coronavirus mouse hepatitis virus strain A59 is not cleaved in primary glial cells and primary hepatocytes.

text: Mouse hepatitis virus strain A59 (MHV-A59) produces meningoencephalitis and severe hepatitis during acute infection. Infection of primary cells derived from the central nervous system (CNS) and liver was examined to analyze the interaction of virus with individual cell types derived from the two principal sites of viral replication in vivo. In glial cell cultures derived from C57BL/6 mice, MHV-A59 produces a productive but nonlytic infection, with no evidence of cell-to-cell fusion. In contrast, in continuously cultured cells, this virus produces a lytic infection with extensive formation of syncytia. The observation of few and delayed syncytia following MHV-A59 infection of hepatocytes more closely resembles infection of glial cells than that of continuously cultured cell lines. For MHV-A59, lack of syncytium formation correlates with lack of cleavage of the fusion glycoprotein, or spike (S) protein. The absence of cell-to-cell fusion following infection of both primary cell types prompted us to examine the cleavage of the spike protein. Cleavage of S protein was below the level of detection by Western blot analysis in MHV-A59-infected hepatocytes and glial cells. Furthermore, no cleavage of this protein was detected in liver homogenates from C57BL/6 mice infected with MHV-A59. Thus, cleavage of the spike protein does not seem to be essential for entry and spread of the virus in vivo, as well as for replication in vitro.


title: Efficacy and safety of pimecrolimus cream in the long-term management of atopic dermatitis in children.

text: OBJECTIVE Pimecrolimus cream (SDZ ASM 981), a nonsteroid inhibitor of inflammatory cytokines, is effective in atopic dermatitis (AD). We assessed whether early treatment of AD signs/symptoms with pimecrolimus could influence long-term outcome by preventing disease flares. METHODS Early intervention with pimecrolimus was compared with a conventional AD treatment strategy (ie, emollients and topical corticosteroids). In this 1-year, controlled, double-blind study, 713 AD patients (2-17 years) were randomized 2:1 to a pimecrolimus-based or conventional regimen. Both groups used emollients for dry skin. Early AD signs/symptoms were treated with pimecrolimus cream or, in the conventional treatment group, vehicle to prevent progression to flares. If flares occurred, moderately potent topical corticosteroids were mandated. The primary efficacy endpoint was ranked flares at 6 months. Safety was monitored clinically, and a skin recall-antigen test was performed at study completion. RESULTS BASELINE CHARACTERISTICS OF THE PATIENTS: The mean age for both groups was approximately 8 years, and the majority of patients had moderate disease at baseline. PATIENT FOLLOW-UP AND EXPOSURE TO STUDY MEDICATION: The mean duration of follow-up (+/-standard error) was 303.7 (+/-5.30) days in the pimecrolimus group and 235.2 (+/-9.40) days in the control group. The discontinuation rate was significantly higher in the control group than in the pimecrolimus group (51.5% vs 31.6% at 12 months), and proportionately more patients with severe or very severe disease discontinued in the control group. The main reason for the higher discontinuation rate in the control group was unsatisfactory therapeutic effect (30.4% vs 12.4%). This resulted in a substantially higher mean number of study medication treatment days in the pimecrolimus group compared with the control group: 211.9 (69.8% of study days) versus 156.0 (66.3% of study days). Of those patients who completed 12 months on study, 14.2% and 7.0% of patients in the pimecrolimus and vehicle groups, respectively, used study medication continuously. EFFICACY Patients in the pimecrolimus group experienced significantly fewer AD flares than those in the control group, according to the primary efficacy analysis on ranked flares of AD (Van Elteren test). The proportion of patients who completed 6 or 12 months with no flares was approximately twice as high in the pimecrolimus group compared with control (61.0% vs 34.2% at 6 months; 50.8% vs 28.3% at 12 months). Fewer flares were observed in the pimecrolimus group regardless of baseline disease severity, so even severe patients derived benefit from the treatment. The analysis of time to first flare showed that treatment with pimecrolimus was associated with a significantly longer flare-free period (log- rank test). Covariate analysis indicated a statistically significant effect on time to first flare of baseline Eczema Area and Severity Index score, and whether patients had "severe" or "very severe" disease at baseline according to the Investigators' Global Assessment, although patients in all baseline disease severity subgroups benefited from treatment. Age had no significant effect. Fewer patients in the pimecrolimus group required topical corticosteroid therapy compared with control (35.0% vs 62.9% at 6 months; 42.6% vs 68.4% at 12 months), and patients in the pimecrolimus group spent fewer days on topical corticosteroid therapy (57.4% vs 31.6% [pimecrolimus vs control, respectively] spent 0 days on topical corticosteroid therapy, 17.1% vs 27.5% 1-14 days, and 25.5% vs 41.0% >14 days over the 12 months of the study). This steroid-sparing effect of pimecrolimus was evident despite pimecrolimus-treated patients being on study longer than patients in the control group. The average proportion of study days spent on second-line corticosteroids was 4.08% in the pimecrolimus group and 9.10% in the control group. Analysis of Eczema Area and Severity Index over time showed significantly lower median scores, thus indicating better disease control in the pimecrolimus group compared with the control group. Similar results were obtained from analysis of the Investigators' Global Assessment (not shown). The treatment groups were well balanced with respect to the number of patients using antihistamines during the study (57.2% vs 62.9%, pimecrolimus vs control, respectively). SAFETY There were no appreciable differences between treatment groups in the overall incidence of adverse events. The most frequent adverse events were common childhood infections and ailments, including nasopharyngitis, headache, and cough. The incidence of suspected drug-related adverse events was not significantly different in the pimecrolimus group (24.7% vs 18.7%--pimecrolimus vs control), and the incidence of serious adverse events was low (8.3% vs 5.2%--pimecrolimus vs control). Life-table analysis of incidence of adverse events revealed no significant differences between the treatment groups, except for cough. Local tolerability was good in both treatment groups. The most common application site reaction reported was sensation of burning (10.5% vs 9.3%--pimecrolimus vs control). There were no major differences between treatment groups in the duration or severity of application site reactions, most of which were mild-to-moderate and transient, occurring within the first week of treatment. Skin infections were reported in both groups. There were no between-group differences in the life-table analysis of time to first occurrence of bacterial skin infections nor in the adjusted incidence of bacterial skin infections. Although there were no significant differences between treatment groups in the incidence of individual viral skin infections, the incidence of grouped viral skin infections (12.4% vs 6.3%--pimecrolimus vs control) showed a slightly higher incidence in the pimecrolimus group. Laboratory values and vital signs showed no significant between-group differences. There were no significant differences between treatment groups in response to recall antigens in those patients who remained on study for 12 months. CONCLUSIONS Treatment of early AD signs/symptoms with pimecrolimus was effective in preventing progression to flares in more than half the patients, reducing or eliminating the need for topical corticosteroids. The benefits were consistently seen at 6 months across important disease severity subgroups and with respect to the various predefined efficacy endpoints. Furthermore, these benefits were sustained for 12 months, providing evidence that long-term treatment with pimecrolimus leads to better control of AD. Treatment with pimecrolimus was well tolerated and was not associated with clinically relevant adverse events compared with the conventional treatment group. The results reported here offer the prospect of effective long-term management of AD with reduced need for topical corticosteroids.


title: Amino acids 1055 to 1192 in the S2 region of severe acute respiratory syndrome coronavirus S protein induce neutralizing antibodies: implications for the development of vaccines and antiviral agents.

text: The spike (S) protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) interacts with cellular receptors to mediate membrane fusion, allowing viral entry into host cells; hence it is recognized as the primary target of neutralizing antibodies, and therefore knowledge of antigenic determinants that can elicit neutralizing antibodies could be beneficial for the development of a protective vaccine. Here, we expressed five different fragments of S, covering the entire ectodomain (amino acids 48 to 1192), as glutathione S-transferase fusion proteins in Escherichia coli and used the purified proteins to raise antibodies in rabbits. By Western blot analysis and immunoprecipitation experiments, we showed that all the antibodies are specific and highly sensitive to both the native and denatured forms of the full-length S protein expressed in virus-infected cells and transfected cells, respectively. Indirect immunofluorescence performed on fixed but unpermeabilized cells showed that these antibodies can recognize the mature form of S on the cell surface. All the antibodies were also able to detect the maturation of the 200-kDa form of S to the 210-kDa form by pulse-chase experiments. When the antibodies were tested for their ability to inhibit SARS-CoV propagation in Vero E6 culture, it was found that the anti-SDelta10 antibody, which was targeted to amino acid residues 1029 to 1192 of S, which include heptad repeat 2, has strong neutralizing activities, suggesting that this region of S carries neutralizing epitopes and is very important for virus entry into cells.


title: Bgp2, a new member of the carcinoembryonic antigen-related gene family, encodes an alternative receptor for mouse hepatitis viruses.

text: Murine coronaviruses such as mouse hepatitis virus (MHV) infect mouse cells via cellular receptors that are isoforms of biliary glycoprotein (Bgp) of the carcinoembryonic antigen gene family (G. S. Dveksler, C. W. Dieffenbach, C. B. Cardellichio, K. McCuaig, M. N. Pensiero, G.-S. Jiang, N. Beauchemin, and K. V. Holmes, J. Virol. 67:1-8, 1993). The Bgp isoforms are generated through alternative splicing of the mouse Bgp1 gene that has two allelic forms called MHVR (or mmCGM1), expressed in MHV-susceptible mouse strains, and mmCGM2, expressed in SJL/J mice, which are resistant to MHV. We here report the cloning and characterization of a new Bgp-related gene designated Bgp2. The Bgp2 cDNA allowed the prediction of a 271-amino-acid glycoprotein with two immunoglobulin domains, a transmembrane, and a putative cytoplasmic tail. There is considerable divergence in the amino acid sequences of the N-terminal domains of the proteins coded by the Bgp1 gene from that of the Bgp2-encoded protein. RNase protection assays and RNA PCR showed that Bgp2 was expressed in BALB/c kidney, colon, and brain tissue, in SJL/J colon and liver tissue, in BALB/c and CD1 spleen tissue, in C3H macrophages, and in mouse rectal carcinoma CMT-93 cells. When Bgp2-transfected hamster cells were challenged with MHV-A59, MHV-JHM, or MHV-3, the Bgp2-encoded protein served as a functional MHV receptor, although with a lower efficiency than that of the MHVR glycoprotein. The Bgp2-mediated virus infection could not be inhibited by monoclonal antibody CC1 that is specific for the N-terminal domain of MHVR. Although CMT-93 cells express both MHVR and Bgp2, infection with the three strains of MHV was blocked by pretreatment with monoclonal antibody CC1, suggesting that MHVR was the only functional receptor in these cells. Thus, a novel murine Bgp gene has been identified that can be coexpressed in inbred mice with the Bgp1 glycoproteins and that can serve as a receptor for MHV strains when expressed in transfected hamster cells.


title: Genotypes of canine distemper virus determined by analysis of the hemagglutinin genes of recent isolates from dogs in Japan.

text: Canine distemper of domestic dogs is caused by canine distemper virus (CDV), a member of the morbilliviruses. It has been a highly contagious disease of great veterinary importance for centuries, but for the last several decades it has been controlled satisfactorily by modified live vaccines. In the 1990s, however, it was described that CDV strains genetically different from vaccine strains may have caused the disease in vaccinated dogs. The highest antigenic variation is found in the H protein. Therefore, in the present study, hemagglutinin (H) genes obtained from current vaccines and field isolates and amplified directly from clinical specimens were genetically analyzed by restriction fragment length polymorphism assay and sequencing. Phylogenetic analysis of the H-gene amino acid sequences revealed that at least two CDV genotypes are circulating among dogs in Japan; one is a genotype to which almost all Japanese CDV isolates belong and the other has not been previously described. Both are separate and independent from the other lineages or genotypes of vaccine strains, as well as European and U.S. CDV isolates. The results suggest that CDV has also evolved in Japan, and further studies will be needed for an evaluation and possible improvement of the efficacies of current CDV vaccines.


title: Twenty years of clinical human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) in Hong Kong.

text: OBJECTIVE To elucidate the development of human immunodeficiency virus (HIV) clinical care and research in Hong Kong. DATA SOURCES Articles on clinical HIV and acquired immunodeficiency syndrome (AIDS) published from 1985 to 2004 were identified through four sources: Red Ribbon Centre, Special Preventive Programme, Secretariat of the Scientific Committee on AIDS, and PubMed search. The first three are operated by the Centre for Health Protection, Department of Health, Hong Kong. STUDY SELECTION Key words for the literature search were 'AIDS', 'HIV', and 'Hong Kong'. DATA EXTRACTION Only papers with original local data were included. DATA SYNTHESIS Sixty papers were identified. The contents were catalogued under seven areas: clinical epidemiology, HIV disease course and presentation, specific complications or organ-based manifestations, immunological evaluation and other monitoring, antiretroviral therapy, HIV/AIDS mortality, and HIV in specific groups. Prevalence of HIV has remained low in Hong Kong but new infections continue to occur together with a significant number of late presenters. Three published AIDS patients' series, up to the first 200 reported cases, identified Pneumocystis carinii pneumonia as the most common AIDS-defining illness in Hong Kong. Penicillium marneffei and Mycobacterium tuberculosis were two important specific infections studied most; uniqueness of the former in patients of South-East Asia was evident. Local studies of Kaposi's sarcoma and HIV-associated lymphoma have also been reported. Research on CD4 counts has revealed that it is lower in healthy and HIV-infected Chinese than their western counterparts. Children, pregnant women, and haemophiliac patients infected with HIV are among the specific groups of patients studied. Survival of patients with advanced disease has greatly improved over the years, particularly after the advent of highly active antiretroviral therapy. CONCLUSION The clinical presentation and outcome of HIV/AIDS patients in Hong Kong are a mixture of those of western and developing countries. Research on clinical HIV/AIDS in Hong Kong is not only beneficial to the planning of patient care, but also enables the formulation of treatment guidelines and provides a reference for other countries.