title: Chemically modified antigen preferentially elicits induction of Th1-like cytokine synthesis patterns in vivo.

text: Differential activation of CD4+ T cell subsets in vivo leads to the development of qualitatively different effector responses. We identify an approach that allows selective activation of strongly Th1-dominated immune responses to protein antigens. Whereas in vivo administration of ovalbumin (OVA) induces cytokine synthesis that is neither Th1 nor Th2 dominated, administration of glutaraldehyde polymerized, high relative molecular weight OVA (OA-POL) leads to 20-fold increase in the ratio of interferon gamma (IFN-gamma)/IL-4 and IFN-gamma/IL-10 synthesis observed after short-term, antigen-mediated restimulation directly ex vivo. In contrast, concurrent in vivo administration of anti-IFN-gamma mAb and OVA or OA-POL results in marked increases in IL-4 and IL-10, and decreased IFN-gamma production, reflecting a polarization of the response towards a Th2-like pattern of cytokine synthesis. These observations may be useful in clinical settings including hypersensitivity, autoimmune diseases, and vaccine development where the ability to actively select specific patterns of cytokine gene expression would be advantageous.


A systematic review of the effect of isolation on psychological wellbeing among adults with an organism of significance* in an acute tertiary referral centre.


title: Induction of alpha interferon by transmissible gastroenteritis coronavirus: role of transmembrane glycoprotein E1.

text: Epithelial cells infected with the coronavirus transmissible gastroenteritis virus (TGEV) and fixed by glutaraldehyde induced a high alpha interferon (IFN-alpha) production in nonimmune porcine as well as human or bovine peripheral blood mononuclear cells (PBMC). IFN-alpha was detected as early as 3 h after exposure of PBMC to infected cells and at producer/inducer cell ratios as low as 1/1. Two of four monoclonal antibodies directed against the viral transmembrane glycoprotein E1 could block the IFN-inducing capacity of both TGEV-infected cells and viral particles. On the other hand, IFN-alpha induction was not markedly affected by monoclonal antibodies directed against other E1 epitopes, against peplomer glycoprotein E2, or against nucleocapsid protein. Thus, these findings strongly imply that IFN induction by TGEV results from interactions between an outer membrane domain of E1 and the PBMC membrane.


Proteolytic cleavage of peplomeric glycoprotein E2 of MHV yields two 90K subunits and activates cell fusion.


title: Prevalence and genetic characterization of human coronaviruses in southern Thailand from July 2009 to January 2011.

text: Emergence of viruses belonging to the coronavirus family has been widespread in the past, causing respiratory infections in humans, such as severe acute respiratory syndrome (SARS). This study investigated the prevalence of human coronavirus (HCoV) and characterized the molecular viral genetics. We collected 1,254 samples from patients diagnosed with respiratory infection in southern Thailand from July 2009 to January 2011 and screened for HCoV by RT-PCR and genotyped by BLAST analysis of nsp12 gene. Phylogenetic analysis was performed based on S gene sequences. Thirty-five of 1,254 samples were positive for HCoV. Viral genotyping revealed 4 genotypes with HCoV-OC43 being the predominant genotype. Viral prevalence and genotype distribution were not in accordance with seasonal distribution. Phylogenetic analysis and deduced amino acid sequences of the S gene showed amino acid variations in each genotype. The S gene sequence of HCoV-OC43 genotype indicated that it resulted from recombination between subgenotypes B and C. Viral genetics analysis disclosed genetic variations of HCoV and additionally, it can provide information suitable for monitoring and prevention of the emergence and re-emergence of various types of coronavirus.


[Severe acute respiratory syndrome].


Genetic association studies in Thorax.


title: Mechanical ventilation in mass casualty scenarios. Augmenting staff: project XTREME.

text: Disaster preparedness typically includes plans that address the need for surge capacity to manage mass-casualty events. A major concern of disaster preparedness in respiratory therapy focuses on responding to a sudden increase in the volume of patients who require mechanical ventilation. Plans for such disasters must include contingencies to address surge capacity in ventilator inventories and the respiratory therapy staff who will manage the ventilators. Tactics to address these situations include efforts to lower demand by transferring patients to other institutions as well as efforts to augment staffing levels. Staff can be augmented by mobilization of deployable teams of volunteers from outside the region and through exploitation of local resources. The latter includes strategies to recruit local respiratory therapists who are currently in either non-clinical or non-hospital-based positions and policies that optimize existing respiratory therapy resources within an institution by canceling elective surgeries, altering shift structure, and postponing vacations. An alternative approach would employ non-respiratory-therapy staff to assist in the management of patients with respiratory failure. Project XTREME (Cross-Training Respiratory Extenders for Medical Emergencies) is a cross-training program developed to facilitate training of non-respiratory-therapy health professionals to assist in the management of patients who require mechanical ventilation. It includes an interactive digital video disc as well as a competency validation laboratory and is designed to be performed at the time of an emergency. Pilot testing of the program suggests it is effective.


title: Human bocavirus.

text: Human bocavirus (HBoV) was first described in 2005 in nasopharyngeal aspirates of children with respiratory tract infection. Multiple studies have confirmed the presence of HBoV in respiratory tract samples of children world-wide. HBoV has recently also been detected in blood and fecal samples. Most studies so far have studied virus prevalence, and only a few reports provide data regarding the linkage of HBoV to disease. These reports indicate that HBoV infection is indeed associated with acute respiratory tract symptoms, but also that HBoV may persist in the respiratory tract for a longer time than other respiratory agents, resulting in frequent detection of low load HBoV carriage. This phenomenon has complicated the use of PCR diagnostics, which has been the only available diagnostic method. Development of alternative diagnostic strategies such as serology will be important for future studies of HBoV and its association with disease.


title: The 3' untranslated region of coronavirus RNA is required for subgenomic mRNA transcription from a defective interfering RNA.

text: The 3'-end of mouse hepatitis virus (MHV) genomic RNA contains a recognition sequence (55 nucleotides [nt]) required for minus-strand RNA synthesis. To determine whether the 3'-end sequence is also involved in subgenomic mRNA transcription, we have constructed MHV defective interfering (DI) RNAs which contain a chloramphenicol acetyltransferase (CAT) gene placed behind an intergenic sequence and a 3'-end sequence with various degrees of internal deletions. The DI RNAs were transfected into MHV-infected cells, and CAT activities, which represent subgenomic mRNA transcription from the intergenic site, were determined. The results demonstrated that the deletions of sequence upstream of the 350 nt at the 3'-end, which include the 3'-untranslated region (3'-UTR), of MHV genomic RNA did not affect subgenomic mRNA transcription. However, deletions that reduced the 3'-end sequences to 270 nt or less completely abolished the mRNA transcription despite the fact that all of these clones synthesized minus-strand RNAs. These results indicated that mRNA transcription from an intergenic site in the MHV DI RNA requires most of the 3'-UTR as a cis-acting signal, which likely exerts its effects during plus-strand RNA synthesis. A substitution of the corresponding bovine coronavirus sequence for the MHV sequence within nt 270 to 305 from the 3'-end abrogated the CAT gene expression, suggesting a very rigid sequence requirement in this region. The deletion of a putative pseudoknot structure within the 3'-UTR also abolished the CAT gene expression. These findings suggest that the 3'-UTR may interact with the other RNA regulatory elements to regulate mRNA transcription.


Dyspnea doesn't always signify bronchial asthma.


title: Coronavirus-induced encephalomyelitis: balance between protection and immune pathology depends on the immunization schedule with spike protein S.

text: The neurotropic mouse hepatitis virus MHV-JHM induces central nervous system (CNS) demyelination in Lewis rats that pathologically resembles CNS lesions in multiple sclerosis. The mechanisms of MHV-JHM-induced demyelination remain unclear and several studies have implicated the role of the immune response in this process. We have shown previously that protective immunity against MHV-JHM-induced encephalomyelitis was induced by immunization with a vaccinia virus (VV) recombinant expressing MHV-JHM S-protein (VV-S). Here, we present evidence that the time of MHV-JHM challenge after immunization with VV-S plays a critical role in protective immunity. The induction of virus-neutralizing S-protein-specific antibodies prior to the MHV-JHM challenge modulates the disease process and a subacute encephalomyelitis based on a persistent virus infection developed. Typical pathological alterations were lesions of inflammatory demyelination. In addition, the results indicate that after seroconversion, CD8+ T cells were no longer essential for virus elimination in contrast to their role in protection during acute encephalomyelitis.


Ethical conduct of research in resource-limited settings.


title: Difference in receptor usage between severe acute respiratory syndrome (SARS) coronavirus and SARS-like coronavirus of bat origin.

text: Severe acute respiratory syndrome (SARS) is caused by the SARS-associated coronavirus (SARS-CoV), which uses angiotensin-converting enzyme 2 (ACE2) as its receptor for cell entry. A group of SARS-like CoVs (SL-CoVs) has been identified in horseshoe bats. SL-CoVs and SARS-CoVs share identical genome organizations and high sequence identities, with the main exception of the N terminus of the spike protein (S), known to be responsible for receptor binding in CoVs. In this study, we investigated the receptor usage of the SL-CoV S by combining a human immunodeficiency virus-based pseudovirus system with cell lines expressing the ACE2 molecules of human, civet, or horseshoe bat. In addition to full-length S of SL-CoV and SARS-CoV, a series of S chimeras was constructed by inserting different sequences of the SARS-CoV S into the SL-CoV S backbone. Several important observations were made from this study. First, the SL-CoV S was unable to use any of the three ACE2 molecules as its receptor. Second, the SARS-CoV S failed to enter cells expressing the bat ACE2. Third, the chimeric S covering the previously defined receptor-binding domain gained its ability to enter cells via human ACE2, albeit with different efficiencies for different constructs. Fourth, a minimal insert region (amino acids 310 to 518) was found to be sufficient to convert the SL-CoV S from non-ACE2 binding to human ACE2 binding, indicating that the SL-CoV S is largely compatible with SARS-CoV S protein both in structure and in function. The significance of these findings in relation to virus origin, virus recombination, and host switching is discussed.


title: Characterization of a Borna disease virus glycoprotein, gp18.

text: Borna disease virus is a nonsegmented negative-strand RNA virus that causes neurologic disease in a wide variety of animal hosts. Here we describe identification and characterization of the first glycoprotein in this viral system. The 18-kDa glycoprotein, gp18, has been purified from infected rat brain. Isolation and microsequencing of this protein allowed identification of a 16.2-kDa open reading frame in the viral antigenome. Lectin binding and endoglycosidase sensitivity assays indicate that gp18 is an unusual N-linked glycoprotein.


title: Comparison of genomic and predicted amino acid sequences of respiratory and enteric bovine coronaviruses isolated from the same animal with fatal shipping pneumonia.

text: The complete genome sequences are reported here of two field isolates of bovine coronavirus (BCoV), which were isolated from respiratory and intestinal samples of the same animal experiencing fatal pneumonia during a bovine shipping fever epizootic. Both genomes contained 31028 nucleotides and included 13 open reading frames (ORFs) flanked by 5'- and 3'-untranslated regions (UTRs). ORF1a and ORF1b encode replicative polyproteins pp1a and pp1ab, respectively, that contain all of the putative functional domains documented previously for the closest relative, mouse hepatitis virus. The genomes of the BCoV isolates differed in 107 positions, scattered throughout the genome except the 5'-UTR. Differences in 25 positions were non-synonymous and were located in all proteins except pp1b. Six replicase mutations were identified within or immediately downstream of the predicted largest pp1a-derived protein, p195/p210. Single amino acid changes within p195/p210 as well as within the S glycoprotein might contribute to the different phenotypes of the BCoV isolates.


title: Etiology of childhood community acquired pneumonia and its implications for vaccination.

text: Pneumonia is an important cause of morbidity and mortality among children throughout the world. Vaccines are available for some organisms, but they are underutilized and/or still in development. To evaluate the potential impact of vaccines, we review studies in which the etiology of childhood community-acquired pneumonia was recorded. In North America and Europe (9 studies), the etiology of pneumonia was established in 62% of studied children (range 43%-88%) by use of noninvasive specific methods for microbiologic diagnosis. The most often identified agents were S. pneumoniae (22%), respiratory syncytial virus (RSV) (20%), Haemophilus influenzae (7%), and Mycoplasma pneumoniae (15%). In Africa and South America (8 studies), bacteria were recovered from 56% (range 32%-68%) of severely ill children studied by lung aspirate. The most often isolated bacteria were Streptococcus pneumoniae (33%) and Haemophilus influenzae (21%). A high percentage of H. influenzae strains were not serotype b. Throughout the world, children requiring hospitalization were most likely to have infection caused by pneumococcus, H. influenzae or RSV. Out patients also had Mycoplasma pneumoniae. Countries in Africa and Asia recorded 2 to 10 times more children with pneumonia (7 to 40/100 annually) than in the USA. Widespread use of pneumococcal and H. influenzae type b conjugate vaccines could reduce the frequency of childhood pneumonia by one-third. Further reduction will require development of non-type b H. influenzae, RSV and M. pneumoniae vaccines. This could result in a > 50% reduction of pneumonia in children. This goal should be sought and achieved as soon as possible.


title: Recent trends on hydrogel based drug delivery systems for infectious diseases.

text: Since centuries, the rapid spread and cure of infectious diseases have been a major concern to the progress and survival of humans. These diseases are a global burden and the prominent cause for worldwide deaths and disabilities. Nanomedicine has emerged as the most excellent tool to eradicate and halt their spread. Various nanoformulations (NFs) using advanced nanotechnology are in demand. Recently, hydrogel and nanogel based drug delivery devices have posed new prospects to simulate the natural intelligence of various biological systems. Owing to their unique porous interpenetrating network design, hydrophobic drug incorporation and stimulus sensitivity hydrogels owe excellent potential as targeted drug delivery systems. The present review is an attempt to highlight the recent trends of hydrogel based drug delivery systems for the delivery of therapeutic agents and diagnostics for major infectious diseases including acquired immune deficiency syndrome (AIDS), malaria, tuberculosis, influenza and ebola. Future prospects and challenges are also described.


title: Canine respiratory viruses.

text: Acute contagious respiratory disease (kennel cough) is commonly described in dogs worldwide. The disease appears to be multifactorial and a number of viral and bacterial pathogens have been reported as potential aetiological agents, including canine parainfluenza virus, canine adenovirus and Bordetella bronchiseptica, as well as mycoplasmas, Streptococcus equi subsp. zooepidemicus, canine herpesvirus and reovirus-1,-2 and -3. Enhancement of pathogenicity by multiple infections can result in more severe clinical forms. In addition, acute respiratory diseases associated with infection by influenza A virus, and group I and II coronaviruses, have been described recently in dogs. Host species shifts and tropism changes are likely responsible for the onset of these new pathogens. The importance of the viral agents in the kennel cough complex is discussed.


title: Quantitative analysis of severe acute respiratory syndrome (SARS)-associated coronavirus-infected cells using proteomic approaches: implications for cellular responses to virus infection.

text: We present the first proteomic analysis on the cellular response to severe acute respiratory syndrome-associated coronavirus (SARS-CoV) infection. The differential proteomes of Vero E6 cells with and without infection of the SARS-CoV were resolved and quantitated with two-dimensional differential gel electrophoresis followed by ESI-MS/MS identification. Moreover isotope-coded affinity tag technology coupled with two-dimensional LC-MS/MS were also applied to the differential proteins of infected cells. By combining these two complementary strategies, 355 unique proteins were identified and quantitated with 186 of them differentially expressed (at least 1.5-fold quantitative alteration) between infected and uninfected Vero E6 cells. The implication for cellular responses to virus infection was analyzed in depth according to the proteomic results. Thus, the present work provides large scale protein-related information to investigate the mechanism of SARS-CoV infection and pathogenesis.


title: New method of laparoscopic incisional hernia repair with double circumferential transfascial sutures.

text: BACKGROUND Although, laparoscopic incisional hernia repair (LIHR) provides an alternative method for managing incisional hernias, the ideal procedure for reducing the incidence of postoperative complications remains unclear. PATIENTS AND METHODS We have developed a new method of LIHR that involves a double transfascial suture and does not require the use of spiral tackers. We performed this procedure consecutively in five patients (four males and one female with a mean age of 65.6 years). We describe our new method of LIHR, and present preliminary clinical results. RESULTS The mean defect size was 26.2 ± 15.8 cm(2), and the mesh size that was used was 121.7 cm(2) in all cases. An occult hernia was found in one patient during laparoscopic observation. The mean operative time was 198.4 ± 49.3 minutes with a blood loss of 12.2 ± 24.6 mL. Postoperative courses were uneventful with a median postoperative hospitalization period of 8 days. No patient required mesh removal and none developed a recurrent hernia during the median follow-up period of 13 months. CONCLUSION Although, larger number of patients and longer follow-up will be required to prove the operative adequacy of our new procedure, it appears to represent a feasible option for LIHR.


title: Experimental rhinovirus infection in volunteers.

text: Experimental viral disease studies in volunteers have clarified many aspects of the pathogenesis of human viral disease. Recently, interest has focused on rhinovirus-associated asthma exacerbations, and new volunteer studies have suggested that airway responsiveness (AR) is enhanced during a cold. For scientific, ethical and safety reasons, it is important to use validated methods for the preparation of a virus inoculum and that the particular virological characteristics and host responses should not be altered. We have prepared a new human rhinovirus (HRV) inoculum using recent guidelines and assessed whether disease characteristics (for example, severity of colds or changes in AR) were retained. Studies were conducted in 25 clinically healthy volunteers using a validated HRV inoculum in the first 17 and a new inoculum in the subsequent eight subjects. Severity of cold symptoms, nasal wash albumin levels and airway responsiveness were measured, and the new inoculum was prepared from nasal washes obtained during the cold. The new inoculum was tested using standard virological and serological techniques, as well as a polymerase chain reaction for Mycoplasma pneumoniae. No contaminating viruses or organisms were detected and the methods suggested were workable. Good clinical colds developed in 20 of the 25 subjects and median symptom scores were similar in the validated and new inoculum groups (18 and 17.5, respectively; p=0.19). All subjects shed virus, and there were no differences noted in viral culture scores, nasal wash albumin and rates of seroconversion in the two groups. Although airway responsiveness increased in both groups (p=0.02 and p=0.05), the degree of change was similar. We have performed experimental rhinovirus infection studies and demonstrated similar clinical disease in two inoculum groups. Amplified airway responsiveness was induced; continuing studies will define the mechanisms and suggest modes of treatment.


title: Cardiovascular safety of dipeptidyl peptidase-4 inhibitors: recent evidence on heart failure.

text: The cardiovascular safety of DPP4 inhibitors as a class, especially in regards to heart failure, has been questioned after the publication of first trials (SAVOR-TIMI 53 and EXAMINE) assessing the cardiovascular risks of DPP4 inhibitors alogliptin and sitagliptin in 2013. Although there were no increased risks in composite cardiovascular outcomes, the SAVOR-TIMI 53 trial reported a 27% increase in hospitalization for heart failure in diabetic patients who received the DPP4 inhibitor saxagliptin. There has been substantial increase in knowledge on the heart failure effect of DPP4 inhibition since 2013. This review will summarize the role of the DPP4/incretin axis in heart failure and discuss the findings from recent large scale clinical trials assessing the effects of DPP4 inhibitors on heart failure.


Annotations.


title: Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of Mycoplasma pneumoniae.

text: A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Mycoplasma pneumoniae was developed and evaluated. The assay specifically amplified only M. pneumoniae sequences, and no cross-reactivity was observed for other Mycoplasma species or respiratory bacterial species. The detection limit for this assay was found to be 2 x 10(2) copies, corresponding to 2-20 colour changing units of M. pneumoniae in 1 h, as observed in a real-time turbidimeter and electrophoretic analysis. The accuracy of the LAMP reaction was confirmed by restriction endonuclease analysis as well as direct sequencing of the amplified product. The assay was applied to 95 nasopharyngeal swab samples collected from patients or from healthy individuals, and compared to a real-time PCR assay in-house. A concordance of 100% was observed between the two assays. The LAMP assay is easy to perform, shows a rapid reaction and is inexpensive. It may therefore be applied in the routine diagnosis of M. pneumoniae infection in the clinical laboratory.


title: [Tuberculosis as an occupational disease].

text: Health workers are exposed to numerous infectious diseases from which the patients they attend are suffering and that have been recognised as occupational infectious diseases. Those most frequently reported are acquired by the haematogenous path, especially hepatitis due to virus B. Numerous infectious diseases are transmitted through airways and until the coming of acute severe respiratory syndrome caused by a coronavirus, tuberculosis was one of the most dreaded. In this paper we analyse the reasons why tuberculosis is considered to be an occupational disease in health personnel; which activities entail most risk and the measures that should be adopted to reduce nosocomial transmission.


title: Severe acute respiratory syndrome among children.

text: OBJECTIVE To study the epidemiologic, clinical, laboratory, and radiologic features, prognostic indicators, and short-term to medium-term outcomes for children with severe acute respiratory syndrome (SARS) and to validate the performance characteristics of a clinical case definition, calculated with respect to SARS-associated coronavirus (SARS-CoV) seroconversion. METHODS Children <18 years of age, from a single-site outbreak, who satisfied a clinical case definition for SARS, with subsequent serologic confirmation, were treated according to a standard protocol and prospectively monitored. RESULTS Forty-four children were included. The median age was 12 years. Forty-two children (95.5%) demonstrated an epidemiologic link. Fever, cough, malaise, coryza, sputum production, headache, myalgia, lymphopenia, and elevated lactate dehydrogenase levels were common presenting features. Radiographic findings were nonspecific, but high-resolution computed tomography of the thorax was an early diagnostic aid. A specific reverse transcription-polymerase chain reaction assay for SARS-CoV yielded positive results for <50% of children. Of 9 children who developed hypoxemia, 8 were treated with methylprednisolone. Of 5 children who received intensive care, 3 required assisted ventilation. All children recovered, and serious adverse events in response to treatment were not observed. The outcomes at 3 to 6 months after disease onset, including exercise tolerance, pulmonary functions, and psychologic status, were favorable. An age of >12 years was associated with methylprednisolone therapy for severe illness. After exclusion of the only infant, an age of >12 years was associated with oxygen requirements. Sore throat, high neutrophil count at presentation, and peak neutrophilia were independent factors predicting severe illness. The clinical case definition demonstrated good sensitivity, specificity, and positive and negative predictive values (97.8%, 92.7%, 88%, and 98.7%, respectively) for diagnostic accuracy. CONCLUSIONS Children are susceptible to SARS-CoV infection. Teenagers resemble adults with respect to disease progression and may develop severe illness. The short-term to medium-term outcomes are good. Sore throat and initial and peak neutrophilia seem to be predictors of severe illness. Our clinical case definition performed well in the epidemic.


title: A suspected canine distemper epidemic as the cause of a catastrophic decline in Santa Catalina Island foxes (Urocyon littoralis catalinae).

text: The island fox (Urocyon littoralis catalinae) population on Santa Catalina Island, California, USA declined precipitously in 1999 with an approximate 95% reduction on their eastern range, an area representing 87% of the island. During this investigation, between October 1999 and April 2000, evidence of live foxes dramatically decreased. The only carcass recovered during the decline succumbed to a co-infection of canine distemper virus (CDV) and toxoplasmosis. Sequence analysis of the viral P gene, derived by polymerase chain reaction, indicated that the virus was closely related to CDV from a mainland USA raccoon (Procyon lotor). Nine of 10 foxes trapped in 1999-2000, on the eastern portion of the island after the decline, had serologic evidence of exposure to CDV, whereas only four of 19 foxes trapped in this region in 1998 had antibodies reactive against CDV. The confirmation of CDV in one deceased fox, evidence of exposure to CDV in east-end foxes in 1999-2000 compared to 1998, and documentation of raccoon introductions to the island, implicates canine distemper as the cause of the population decline.


News from the Internet.


title: Tweaking Innate Immunity: the Promise of Innate Immunologicals As Anti-infectives.

text: New and exciting insights into the importance of the innate immune system are revolutionizing our understanding of immune defense against infections, pathogenesis, and the treatment and prevention of infectious diseases. The innate immune system uses multiple families of germline-encoded pattern recognition receptors (PRRs) to detect infection and trigger a variety of antimicrobial defense mechanisms. PRRs are evolutionarily highly conserved and serve to detect infection by recognizing pathogen-associated molecular patterns that are unique to microorganisms and essential for their survival. Toll-like receptors (TLRs) are transmembrane signalling receptors that activate gene expression programs that result in the production of proinflammatory cytokines and chemokines, type I interferons and antimicrobial factors. Furthermore, TLR activation facilitates and guides activation of adaptive immune responses through the activation of dendritic cells. TLRs are localized on the cell surface and in endosomal/lysosomal compartments, where they detect bacterial and viral infections. In contrast, nucleotide-binding oligomerization domain proteins and RNA helicases are located in the cell cytoplasm, where they serve as intracellular PRRs to detect cytoplasmic infections, particularly viruses. Due to their ability to enhance innate immune responses, novel strategies to use ligands, synthetic agonists or antagonists of PRRs (also known as 'innate immunologicals') can be used as stand-alone agents to provide immediate protection or treatment against bacterial, viral or parasitic infections. Furthermore, the newly appreciated importance of innate immunity in initiating and shaping adaptive immune responses is contributing to our understanding of vaccine adjuvants and promises to lead to improved next-generation vaccines.


title: Structure-based discovery of a novel angiotensin-converting enzyme 2 inhibitor.

text: Angiotensin-converting enzyme 2 (ACE2) is considered an important therapeutic target for controlling cardiovascular diseases and severe acute respiratory syndrome (SARS) outbreaks. Recently solved high-resolution crystal structures of the apo-bound and inhibitor-bound forms of ACE2 have provided the basis for a novel molecular docking approach in an attempt to identify ACE2 inhibitors and compounds that block SARS coronavirus spike protein-mediated cell fusion. In this study, approximately 140 000 small molecules were screened by in silico molecular docking. In this structure-activity relation study, the molecules with the highest predicted binding scores were identified and assayed for ACE2 enzymatic inhibitory activity and for their ability to inhibit SARS coronavirus spike protein-mediated cell fusion. This approach identified N-(2-aminoethyl)-1 aziridine-ethanamine as a novel ACE2 inhibitor that also is effective in blocking the SARS coronavirus spike protein-mediated cell fusion. Thus, the molecular docking approach resulting in the inhibitory capacity of N-(2-aminoethyl)-1 aziridine-ethanamine provides an attractive small molecule lead compound on which the development of more effective therapeutic agents could be developed to modulate hypertension and for controlling SARS infections.


Working with Medecins-Sans-Frontieres in China: a personal account.


title: Structural and functional analyses of the severe acute respiratory syndrome coronavirus endoribonuclease Nsp15.

text: The severe acute respiratory syndrome (SARS) coronavirus encodes several RNA-processing enzymes that are unusual for RNA viruses, including Nsp15 (nonstructural protein 15), a hexameric endoribonuclease that preferentially cleaves 3' of uridines. We solved the structure of a catalytically inactive mutant version of Nsp15, which was crystallized as a hexamer. The structure contains unreported flexibility in the active site of each subunit. Substitutions in the active site residues serine 293 and proline 343 allowed Nsp15 to cleave at cytidylate, whereas mutation of leucine 345 rendered Nsp15 able to cleave at purines as well as pyrimidines. Mutations that targeted the residues involved in subunit interactions generally resulted in the formation of catalytically inactive monomers. The RNA-binding residues were mapped by a method linking reversible cross-linking, RNA affinity purification, and peptide fingerprinting. Alanine substitution of several residues in the RNA-contacting portion of Nsp15 did not affect hexamer formation but decreased the affinity of RNA binding and reduced endonuclease activity. This suggests a model for Nsp15 hexamer interaction with RNA.


title: [Effect of integrative Chinese and western medicine on T-lymphocyte subsets in treating patients with severe acute respiratory syndrome].

text: OBJECTIVE To investigate the therapeutic effect of integrative Chinese and western medicine (ICWM) in treating severe acute respiratory syndrome (SARS) and its influence on T-lymphocyte subsets. METHODS Comparative study was conducted in 133 SARS inpatients in Beijing Ditan Hospital, who were divided into 3 groups according to the treatment applied, the basic treated group, the low dose steroid group and the high dose steroid group, and all the 3 groups were subdivided into two groups, Chinese herbs and non-Chinese herbs added, respectively. Chinese drugs for clearing-up heat, dispelling dampness, detoxication, removing stasis, supplementing Qi and nourishing Yin were selected according to patients' syndrome and given additionally to all the three ICWM groups. Retrospective analysis for significance test on changes of T-lymphocyte subsets before and after treatment were carried out. RESULTS T-lymphocyte counts, including CD3+, CD4+, CD8+, lowered in all patients before treatment, but increased significantly after treated for 3 weeks, the increment in all the low dose steroid treated groups was higher than that in the basic treated groups, and that in ICWM groups was higher than that in non-ICWM groups, respectively. CONCLUSION Glycocorticosteroid and Chinese herbal medicine treatments could promote the recovery of T-lymphocyte profile, rationally use of them is the effective therapeutic method.


title: Epidemiological surveillance and control of rubella in Singapore, 1991-2007.

text: INTRODUCTION We reviewed the epidemiological features of rubella in Singapore and the impact of the national immunisation programme in raising the population herd immunity against rubella, with special reference to females in the reproductive age group, and in the elimination of congenital rubella syndrome (CRS). MATERIALS AND METHODS Epidemiological data on all reported cases of rubella and CRS were obtained from the Communicable Diseases Division and Central Claims Processing System, respectively, at the Ministry of Health. Coverage of the childhood immunisation programme against rubella was based on the immunisation data maintained by the National Immunisation Registry, Health Promotion Board. To assess the herd immunity of the population against rubella, 4 serological surveys were conducted from 1989 to 1990, in 1993, 1998 and 2004. RESULTS The incidence of rubella has decreased significantly from the peak of 13.3 per 100,000 population in 1996 to 1.8 per 100,000 in 2007. CRS has virtually disappeared. With more than 92% to 93% of primary school leavers and preschool children annually vaccinated against rubella since 1976 and 1990, respectively, the level of susceptibility to rubella among women in the reproductive age group has gradually decreased from 44% in 1975 to 28% in 1985, and maintained at between 10% and 20% from 1987 to 1998. A considerable proportion (15.8%) of women 18 to 44 years of age remained susceptible to rubella infection in 2004. CONCLUSION Rubella prevention and control has been successfully implemented. However, the relatively high level of susceptibility to rubella among women in the reproductive age group continues to be of concern. More public awareness and health educational efforts are needed and every opportunity should be taken to ensure that all susceptible women are identified and protected against the infection.


title: Senescent BALB/c mice are able to develop resistance to Leishmania major infection.

text: Aging has been associated with a decline in immunocompetence and resistance to infections, partially due to dysregulated NO production by macrophages and deficits in mounting Th2 cell responses. We wondered if these alterations would reverse the immune response in experimental leishmaniasis. Bone-marrow-derived macrophages from 2- and 18-month-old (senescent) C57BL/6 or BALB/c mice showed no marked difference in leishmanicidal functions. In vivo infections of resistant C57BL/6 mice with Leishmania major revealed no difference between senescent and young mice. However, among susceptible BALB/c mice, senescent animals showed less foot-pad swelling than young mice, and 40 to 60% of them even showed healing of ulcers, reduced parasite dissemination, and a Th1 cell response. These changes were associated with a spontaneous release of interleukin-12 (IL-12) by macrophages from aged but not from young mice. Since exogenous microbial stimulation can influence immune responses during aging, we also infected senescent mice who were raised under specific-pathogen-free (SPF) conditions. They showed neither resistance nor a Th1 response, but their macrophages still spontaneously released IL-12. A microbiological analysis showed that conventionally kept mice, but not SPF mice, had experienced infection with murine hepatitis virus (MHV), an infection associated with a Th1-like response. We conclude that for the reversal of the immune response, senescence is the premier requirement but needs to be completed by another mandatory event such as microbial stimulation. One of the age-related, but not environment-related, factors is the spontaneous release of IL-12 by macrophages, while confrontation with MHV presents an environment-related difference, with both having the potential to support a Th1 response.


COVID-19: Main therapeutic options.


title: [Serum antibodies detection for serological diagnosis of severe acute respiratory syndrome].

text: OBJECTIVE To evaluate the significance of serum specific antibodies against severe acute respiratory syndrome (SARS) virus in the diagnosis of SARS and the concordance of IFA and ELISA in the antibodies measurements. METHODS Specific antibodies of IgM and IgG against SARS virus in 267 serum specimens from patients with SARS and 132 from non-SARS patients or healthy persons were assayed with IFA or ELISA. The sensitivity, specificity, positive predictive value (ppv), negative predictive value (NPV), and accuracy were used to evaluate the significance of the antibodies in diagnosis of SARS. The concordance of different measurements was tested with Kappa. RESULTS The positive rate of specific IgM and IgG detected by IFA increased sharply on the eleventh day of the disease. The sensitivity, specificity, PPV, NPV, and accuracy of IgM in diagnosis of SARS on eleventh day or later were 65.6%, 100.0%, 100.0%, 71.0% and 81.3%, respectively, and IgG were 91.1%, 97.0% 97.3%, 90.1%, and 93.8%, respectively. Similar results could be seen when IgM and IgG were detected by ELISA. The Kappa of IFA and ELISA for IgM and IgG were 0.640 and 0.779 respectively. CONCLUSIONS The antibodies against SARS virus can be detected in the mid-and late stage of SARS. The antibodies can be used in the serological diagnosis of SARS. There is high concordance of IFA and ELISA results in measurement of SARS antibodies.


title: Rapidly fatal bacteremic pneumonia caused by Klebsiella pneumoniae with K1 hypermucoviscosity phenotype in a previously healthy young man receiving levofloxacin treatment.

text: Fatal bacteremic Klebsiella pneumoniae pneumonia is commonly encountered in alcoholic and diabetic patients. This report describes a previously healthy young man with rapidly fatal bacteremic pneumonia caused by K. pneumoniae serotype K1, complicated by septic shock and multiple organ dysfunction.


title: Bridges to sustainable tropical health.

text: Ensuring sustainable health in the tropics will require bridge building between communities that currently have a limited track record of interaction. It will also require new organizational innovation if much of the negative health consequences of large-scale economic development projects are to be equitably mitigated, if not prevented. We focus attention on three specific contexts: (i) forging linkages between the engineering and health communities to implement clean water and sanitation on a broad scale to prevent reworming, after the current deworming-only programs, of people by diverse intestinal parasites; (ii) building integrated human and animal disease surveillance infrastructure and technical capacity in tropical countries on the reporting and scientific evidence requirements of the sanitary and phytosanitary agreement under the World Trade Organization; and (iii) developing an independent and equitable organizational structure for health impact assessments as well as monitoring and mitigation of health consequences of economic development projects. Effective global disease surveillance and timely early warning of new outbreaks will require a far closer integration of veterinary and human medicine than heretofore. Many of the necessary surveillance components exist within separate animal- and human-oriented organizations. The challenge is to build the necessary bridges between them.


title: Electrophysiological Biomarkers in Spinal Muscular Atrophy: Preclinical Proof of Concept.

text: OBJECTIVE Preclinical therapies that restore survival motor neuron (SMN) protein levels can dramatically extend survival in spinal muscular atrophy (SMA) mouse models. Biomarkers are needed to effectively translate these promising therapies to clinical trials. Our objective was to investigate electrophysiological biomarkers of compound muscle action potential (CMAP), motor unit number estimation (MUNE) and electromyography (EMG) using an SMA mouse model. METHODS Sciatic CMAP, MUNE, and EMG were obtained in SMNΔ7 mice at ages 3-13 days and at 21 days in mice with SMN selectively reduced in motor neurons (ChATCre ). To investigate these measures as biomarkers of treatment response, measurements were obtained in SMNΔ7 mice treated with antisense oligonucleotide (ASO) or gene therapy. RESULTS CMAP was significantly reduced in SMNΔ7 mice at days 6-13 (p<0.01), and MUNE was reduced at days 7-13 (p<0.01). Fibrillations were present on EMG in SMNΔ7 mice but not controls (p=0.02). Similar findings were seen at 21 days in ChATCre mice. MUNE in ASO-treated SMNΔ7 mice were similar to controls at day 12 and 30. CMAP reduction persisted in ASO-treated SMNΔ7 mice at day 12 but was corrected at day 30. Similarly, CMAP and MUNE responses were corrected with gene therapy to restore SMN. INTERPRETATION These studies confirm features of preserved neuromuscular function in the early postnatal period and subsequent motor unit loss in SMNΔ7 mice. SMN restoring therapies result in preserved MUNE and gradual repair of CMAP responses. This provides preclinical evidence for the utilization of CMAP and MUNE as biomarkers in future SMA clinical trials.


title: The relationship between the occurrence of undifferentiated bovine respiratory disease and titer changes to Haemophilus somnus and Mannheimia haemolytica at 3 Ontario feedlots.

text: The association between exposure to Haemophilus somnus and Mannheimia haemolytica (formerly Pasteurella haemolytica) and the risk of undifferentiated bovine respiratory disease (UBRD) was investigated using serological evidence of exposure coupled with a factorial design vaccine field trial. Measures of previous exposure (titer at arrival) and current exposure (titer increase in the study period) to these agents were used. The vaccine field trial involved systematic allocation of animals into groups that received either a M. haemolytica vaccine, an H. somnus vaccine, a combined M. haemolytica and H. somnus vaccine, and an unvaccinated control group. Serum was collected from the 852 animals enrolled to determine titers to H. somnus, M. haemolytica, bovine coronavirus and bovine viral diarrhea virus. Vaccination with H. somnus in combination with M. haemolytica and with M. haemolytica alone reduced the risk of UBRD. The odds ratio for vaccination with H. somnus alone and UBRD risk suggested some sparing effect, but the 95% confidence limits included unity. There was no association between serological evidence of concurrent exposure to M. haemolytica and UBRD occurrence. There was an association between titer change to H. somnus and UBRD risk. However, the association changed with time of BRD treatment; animals diagnosed and treated for UBRD on or after day 10 showed little evidence of exposure to H. somnus, despite evidence of natural H. somnus exposure in the unvaccinated group. The association between titer change to H. somnus and UBRD occurrence seen in this study may be a consequence of prolonged exposure to antibiotics, rather than a causal association.


title: Enteropathogens associated with acute diarrhoeal diseases.

text: Five types of Escherichia coli are responsible for as much as 25% of all diarrheal diseases in developing countries. They tend to be transmitted via contaminated foods, particularly weaning foods, and water. They include enterotoxigenic, enteropathogenic, enteroadherent, enteroinvasive, and enterohemorrhagic E. coli. Shigella species are responsible for 10-15% of acute diarrheas in children less than 5 years old and the most common etiologic agents of childhood dysentery. Shigellosis is common in the warm season. An outbreak of shigella dysentery in West Bengal, India, had a high attack rate in children less than 5 years old and was resistant to many drugs. Nontyphoid Salmonella species cause watery diarrhea with nausea, cramps, and fever. Worldwide, various Salmonella strains exhibit resistance to ampicillin, chloramphenicol, and co-trimoxazole. Campylobacter jejuni produces watery diarrhea which, in 33% of cases and 1-2 days after onset, contains blood and mucus. Many normal healthy children in developing countries are carriers of C. jejuni. Vibrio cholerae O1 is endemic in parts of Africa and Asia (e.g., 5-10% of hospitalized diarrhea patients). The ElTor cholera biotype is responsible for the 7th pandemic. Other bacterial enteropathogens are Aeromonas species, Bacteroides fragilis, and Providencia alcalifaciens. Rotavirus is a major cause of sporadic and epidemic diarrhea among 6-23 month olds. Its incidence peaks in cold or dry seasons. Other viral enteropathogens are Norwalk virus, adenoviruses, astroviruses, and coronaviruses. In India, the prevalence of Entamoeba histolytica varies from 3.6% to 47.4%. It occurs equally in high and low socioeconomic classes. Giardia lamblia usually infects 1-5 year old children. Its transmission routes are food, water, and the fecal-oral route. Cryptosporidia produce acute watery diarrhea, especially in children less than 2 years old. Cryptosporidia diarrhea is common among AIDS patients. Oral rehydration therapy and proper feeding during and after diarrhea reduces deaths from diarrhea.


Case scenario: perioperative administration of tocotrienols and green tea extract in a child with familial dysautonomia.


title: Oral immunoadjuvant activity of Lactobacillus casei subsp. casei in dextran-fed layer chickens.

text: We recently reported that synbiotic Lactobacillus casei subsp. casei together with specific substrate dextran elicited an enhancement in humoral immune response against bovine serum albumin (BSA) as a model antigen in BALB/c mice. The present study was designed to evaluate the oral immunoadjuvant effects of the synbiotic in layer chickens. Using a PCR assay, L. casei subsp. casei was detected specifically in the intestinal chyme of chickens (10 d of age, Julia strain) fed ad libitum on a diet supplemented with 75 mg dextran/kg (dextran-supplemented diet, DSD) and administered orally with 10(7) colony-forming units (CFU) L. casei subsp. casei in 0.1 ml PBS with the aid of an intubation needle at 1, 2 and 3 d of age. Furthermore, oral administration of 10(7) CFU L. casei subsp. casei at 1-3 d of age significantly enhanced the production of anti-BSA antibody in DSD-fed chickens (60 d of age) administered orally with 1 mg BSA at 32 and 33 d of age and subcutaneously with 5 microg BSA at 33 d of age. In addition, among bacterial numbers tested, 10(6) CFU L. casei subsp. casei together with dextran induced an effective increase in humoral immune response to mixed inactivated vaccines against Newcastle disease and avian infectious bronchitis, and the treatment may be advantageous in protecting against these infectious diseases in chickens in actual application. These results suggest that dietary supplementation of L. casei subsp. casei with dextran leads to immunomodulation of humoral immune responses.


title: Almanac 2013: novel non-coronary cardiac interventions.

text: Recent innovations in interventional cardiology have dramatically expanded the therapeutic options for patients with cardiac conditions. Interventional cardiology is no longer limited to the treatment of coronary artery disease but allows also treatment of valvular disease, stroke prevention, hypertension, etc. One of the most important new treatment options is the percutaneous treatment for aortic valve stenosis (transcatheter aortic valve implantation), since aortic valve disease is a rather common problem in elderly patients, with many of them at high risk for surgery. Similarly, mitral regurgitation is often associated with comorbidities which make surgery high risk. The MitraClip is a promising percutaneous alternative to surgical valve repair or replacement. Other procedures discussed in this review are the percutaneous left atrial appendage closure as a non-pharmacologic therapy to prevent strokes, and renal denervation for resistant hypertension. This review explains the basic principles of these procedures, the most important clinical evidence, and also provides additional recent clinical data on each of them.


title: Long-term management of vaccine-induced refractory ischemic dermatopathy in a Miniature Pinscher puppy.

text: A 2-month-old intact female Miniature Pinscher puppy presented with footpad swelling and crusted pustules of ear pinnae. The dog had been vaccinated with a polyvalent canine vaccine 5 days prior to the onset of clinical signs. With the history of recent vaccination, the clinical presentation and the histopathological observations were suggestive of ischemic dermatopathy. Treatment involved oral prednisolone, azathioprine, and other immune modulating drugs, which did not work. Chlorambucil plus cyclosporine therapy was initiated for vigorous immune suppression after rush therapy using intravenous immunoglobulin. Clinical signs again gradually improved with no relapse or side effects, even at a 4-month follow-up. The case report is suggested ischemic dermatopathy refractory to conventional therapy and suggests effective approaches to long-term management of the disease.


title: Pedagogy for teaching and learning cooperatively on the Web: a Web-based pharmacology course.

text: The Internet is becoming a preferred place to find information. Millions of people go online in the search of health and medical information. Likewise, the demand for Web-based courses grows. This article presents the development, utilization and evaluation of a web-based pharmacology course for nursing students. The course was developed based on 150 commonly used drugs. There were 110 year 1 nursing students took part in the course. After attending six hours face to face lecture of pharmacology over three weeks, students were invited to complete a questionnaire (pre-test) about learning pharmacology. The course materials were then uploaded to a WebCT for student's self-directed learning and attempts to pass two scheduled online quizzes. At the end of the semester, students were given the same questionnaire (post-test). There were a significant increase in the understanding compared with memorizing the subject content, the development of problem solving ability in learning pharmacology and becoming an independent learner (p ,0.05). Online quizzes yielded satisfactory results. In the focused group interview, students appreciated the time flexibility and convenience associated with web-based learning, also, they had made good suggestions in enhancing web-based learning. Web-based approach is promising for teaching and learning pharmacology for nurses and other health-care professionals.


title: eIF3-f function in skeletal muscles: to stand at the crossroads of atrophy and hypertrophy.

text: The control of muscle cell size is a physiological process balanced by a fine tuning between protein synthesis and protein degradation. MAFbx/Atrogin-1 is a muscle specific E3 ubiquitin ligase upregulated during disuse, immobilization and fasting or systemic diseases such as diabetes, cancer, AIDS and renal failure. This response is necessary to induce a rapid and functional atrophy. To date, the targets of MAFbx/Atrogin-1 in skeletal muscle remain to be identified. We have recently presented evidence that eIF3-f, a regulatory subunit of the eukaryotic translation factor eIF3 is a key target that accounts for MAFbx/Atrogin-1 function in muscle atrophy. More importantly, we showed that eIF3-f acts as a "translational enhancer" that increases the efficiency of the structural muscle proteins synthesis leading to both in vitro and in vivo muscle hypertrophy. We propose that eIF3-f subunit, a mTOR/S6K1 scaffolding protein in the IGF-1/Akt/mTOR dependent control of protein translation, is a positive actor essential to the translation of specific mRNAs probably implicated in muscle hypertrophy. The central role of eIF3-f in both the atrophic and hypertrophic pathways will be discussed in the light of its promising potential in muscle wasting therapy.


Inadvertent catheterization of the ascending lumbar vein.


Biological and molecular characteristics of an HEV isolate associated with recent acute outbreaks of encephalomyelitis in Quebec pig farms.


title: Class I and class II viral fusion protein structures reveal similar principles in membrane fusion.

text: Recent crystal structures of Flavivirus and Alphavirus fusion proteins (class II) confirm two major principles of protein machineries that mediate the merger of two opposing lipid bilayers. First, the fusion protein can bridge both membranes tethered by two membrane anchors. Second, refolding or domain rearrangement steps lead to the positioning of both anchors into close proximity at the same end of an elongated structure. Although these two steps are in principle sufficient to pull two opposing membranes together and initiate membrane fusion, accumulating evidence suggests that the process requires the concerted action of a number of fusion proteins at and outside the contact sites. This review will focus on the structures of viral class I and class II fusion proteins and their similarities in facilitating membrane fusion.


Lessons from SARS: challenges for the international nursing research community.


title: [Sero-epidemiological characteristics of antibody to SARS-associated coronavirus in Gansu Province].

text: OBJECTIVE To study the levels of SARS-associated coronavirus IgG antibody of SARS patients, people who closely contacted SARS patients and normal subjects in Gansu province. METHODS The levels of SARS-associated coronavirus IgG antibody were measured by ELISA. The material included acute and (or) recovery period sera of 9 SARS patients, sera from 1,109 doctors and nurses closely contacted with SARS patients, laboratory workers, personnel for disease control and prevention, persons who contacted SARS patients, and sera from 978 normal subjects. RESULTS SARS coronavirus IgG antibody was detected positive in 6 of the 9 patients, it was still positive in the sera twelve months after recovery; 1 of the closely contacted persons and 3 normal subjects were found positive. CONCLUSION The positive rate of SARS coronavirus IgG antibody of patients was consistent with the clinical diagnosis. The low positive rate of the persons who closely contacted SARS patients and normal subjects suggests that SARS probably had no subclinical infection.


title: Generation of a defective RNA of avian coronavirus infectious bronchitis virus (IBV). Defective RNA of coronavirus IBV.

text: The Beaudette strain of IBV was passaged 16 times in chick kidney (CK) cells. Total cellular RNA was analyzed by Northern hybridization and was probed with 32P-labeled cDNA probes corresponding to the first 2 kb of the 5' end of the genome, but excluding the leader, and to the last 1.8 kb of the 3' end of the genome. A new, defective IBV RNA species (CD-91) was detected at passage six. The defective RNA, present in total cell extract RNA and in oligo-(dT)30-selected RNA from passage 15, was amplified by the reverse transcription-polymerase chain reaction (RT-PCR) to give four fragments. The oligonucleotides used were selected such that CD-91 RNA, but not the genomic RNA, would be amplified. Cloning and sequencing of the PCR products showed that CD-91 comprises 9.1 kb and has three regions of the genome. It contains 1133 nucleotides from the 5' end of the genome, 6322 from gene 1b corresponding to position 12423 to 18744 in the IBV genome and 1626 from the 3' end of the genome. At position 749 one nucleotide, an adenine residue, was absent from CD-91 RNA. By Northern hybridization CD-91 RNA was detected in virions in higher amounts than the subgenomic mRNAs.


title: Membrane fusion.

text: Fusion of biological membranes is governed by physical principles but it is unclear whether the transition states are primarily determined by lipid physics or by protein-lipid interactions. Recent advances in the field include the physical description of bilayer fusion, particularly new models beyond continuum models and the role of the SNARE proteins. Despite substantial progress, an integrated concept for protein-mediated membrane fusion is not yet available, and many open questions remain to be answered.


title: Peptide domain involved in the interaction between membrane protein and nucleocapsid protein of SARS-associated coronavirus.

text: Severe acute respiratory syndrome (SARS) is an emerging infectious disease associated with a novel coronavirus (CoV) that was identified and molecularly characterized in 2003. Previous studies on various coronaviruses indicate that protein-protein interactions amongst various coronavirus proteins are critical for viral assembly and morphogenesis. It is necessary to elucidate the molecular mechanism of SARS-CoV replication and rationalize the anti-SARS therapeutic intervention. In this study, we employed an in vitro GST pull-down assay to investigate the interaction between the membrane (M) and the nucleocapsid (N) proteins. Our results show that the interaction between the M and N proteins does take place in vitro. Moreover, we provide an evidence that 12 amino acids domain (194-205) in the M protein is responsible for binding to N protein. Our work will help shed light on the molecular mechanism of the virus assembly and provide valuable information pertaining to rationalization of future anti-viral strategies.


title: Determinants of outbreaks of emerging infectious diseases : the case of Chikungunya in Indian ocean, 2004-2007.

text: Emerging outbreaks of arboviroses are most often tied to the convergence of a set of factors which have physical and environmental, genetic and biological, ecological, social, political and economic origins. Thirteen out of these factors are known as main determinants of emerging infectious disease outbreaks. This paper assesses the role of these factors from exploring the Chikungunya outbreak which occurred in Indian Ocean in 2004-2007, as case study. Chikungunya is an alphavirus borne and transmitted by Aedes aegypti and Aedes albopictus.


A candidate vaccine for severe acute respiratory syndrome.


title: [Identification and molecular cloning and sequence analysis of a novel coronavirus from patients with SARS by RT-PCR].

text: OBJECTIVE To investigate the etiologic agents of the SARS and develop diagnostic method for this disease. METHODS Thirty-six nasopharyngeal aspirate specimens from 27 patients with SARS in Shenzhen were collected. The samples were aliquotted to three parts and subjected to molecular assays for human metapneumovirus, chlamydia and a novel coronavirus, which was reported recently to be the etiologic agent of SARS. Nested RT-PCR was used to amplify the RNA polymerase gene of the novel coronavirus and the PCR products were sequenced directly or after cloned to pMD18-T vector. RESULTS Human metapneumovirus and chlamydia genes were detected in none of the specimens using the RT-PCR and nested-PCR, respectively. The novel coronavirus gene were amplified in 6 of 36 specimens, the sequence analysis indicated that this novel coronavirus is unrelated to any other coronavirus reported previously. The nucleotide and deduced amino acid alignment between this coronavirus and others was not more than 40% and 70% to 82%, respectively, while the nucleotide sequence cloned from the 6 patients were identical. CONCLUSIONS The SARS patients in Shenzhen were infected with coronavirus and this novel coronavirus is associated with SARS. The sequence analysis indicated that the coronavirus from SARS patients in Shenzhen is the same as that identified from other areas such as Canada and Hong Kong. A specific diagnostic nested RT-PCR was developed to identify this novel coronavirus infection.


Hemoptysis as an Initial Presentation of COVID-19 - An Observation.


title: [Microbiological study results (bacteriology and electron microscopy) of diarrhea in dog whelps].

text: Bacteriological examinations of 159 faeces and intestinal contents of dogs with diarrhoea revealed E. coli in 157 specimens. 73 of these samples contained non haemolytic strains, 18 haemolytic isolates, and 66 haemolytic, as well as non haemolytic strains. Klebsiella sp. and Staphylococcus aureus were found in 9 cases, and Salmonella sp. (group B) was isolated once only. By electron microscopy parvovirus could be detected in 19 samples. Ten were positive for coronavirus and one for rotavirus. Morphologically not finally identified coronavirus like and picornavirus like particles were found in 3 cases, respectively. A significant relationship between the occurrence of virus infections and the isolation of certain bacteria species was not found. In 45 E. coli strains virulence factors, such as the heat labile enterotoxin (LT) and the verotoxin (VT), could not be detected, but 31 of these isolates showed different haemagglutination patterns which were still present by 14 of them in the presence of mannose (mannose resistant haemagglutination, MR-HA). These MR-HA inducing E. coli isolates were present more often in parvovirus positive samples (in 6 of 10) than they could be detected in parvovirus negative ones (in 8 of 35), implicating a potential pathogenic role of these E. coli strains for the parvovirus enteritis of dogs.


title: Does 11.5 mm guided single port surgery has clinical advantage than multi-port thoracoscopic surgery in spontaneous pneumothorax?

text: BACKGROUND Video-assisted thoracoscopic surgery (VATS) has been widely used for spontaneous pneumothorax (SP). In recent years, thoracic surgeons have attempted single incision or single port surgery with the development of surgical technology and skills. Theoretically, single port surgery is expected to provide benefits such as less pain and early recovery. The purpose of this study was to determine the benefits of single port surgery in SP. METHODS The 107 patients with SP who underwent surgery, between July 2013 and May 2015, were reviewed retrospectively. The patients with secondary pneumothorax, who underwent open procedures and lacking of medical records were excluded. Visual analog scale (VAS), paresthesia and clinical outcomes were reviewed in 86 patients (46 patients: three-port, 40 patients: 11.5 mm guided single-port). RESULTS The mean age was 23.4 years in three-port and 22.4 in single-port (P=0.247). The height and body weight were not significantly difference between two groups. The mean operation time was 39 minutes (mins) in the three-port and 37.3 mins in the single port without statistical difference (P=0.204). The pain score in the single port surgery was significantly lower after postoperative day (POD) 1 (P=0.028). However chest tube duration time was significantly shorter in the single port group (P<0.001). After exclusion of the patients with chest tube removal within postoperative 1 day, the pain score was not significantly different at the POD 1 between two groups (P=0.176). The pain score between two groups were not different at 1 week after discharge. CONCLUSIONS The pain score reduction was found 1 day after operation in the single port group. However, the chest tube duration time was significantly shorter in the single port group and the pain score was not different at 1 week after discharge. Considering young age in primary SP, the benefit of single port surgery in SP was minimal.


title: Morbillivirus infection in a wild siberian tiger in the Russian Far East.

text: We report the first documented case of morbillivirus infection in a wild, free-ranging Siberian tiger (Panthera tigris altaica). The tigress entered a small village in the Russian Far East in an ambulatory but stuporous state with no apparent recognition or fear of humans. Her condition progressed rapidly with neurological signs, anorexia, and ultimately death. Histologic lesions included vacuolated to malacic white matter in the brain stem, cerebellum, and thalamus, with associated lymphocytic meningoencephalitis. Large, intranuclear, eosinophilic inclusions were within regional astrocytes, and the brain lesions were immunohistochemically positive when stained for canine distemper viral antigen. Hematologic and blood chemistry results were consistent with overwhelming systemic infection and starvation. The animal also was antibody-positive for canine distemper virus, feline panleukopenia, and feline coronavirus.


title: Transmissible proventriculitis in broilers.

text: This article reviews transmissible proventriculitis in poultry from 1971 to 2006. The disease is important in commercial broilers worldwide, resulting in reduced profits. The aetiology of this disease is unknown and different clinical presentations often result in a confused or complicated diagnosis. The lesion of enlarged proventriculus is often referred to as proventriculitis. However, the term proventriculitis can only be used correctly when there is microscopic evidence of inflammation of the proventriculus glands. Infectious and non-infectious causes of proventriculitis, with major emphasis on the infectious or transmissible causes, are reviewed.


title: Challenges and limitations of patient-specific vascular phantom fabrication using 3D Polyjet printing.

text: Additive manufacturing (3D printing) technology offers a great opportunity towards development of patient-specific vascular anatomic models, for medical device testing and physiological condition evaluation. However, the development process is not yet well established and there are various limitations depending on the printing materials, the technology and the printer resolution. Patient-specific neuro-vascular anatomy was acquired from computed tomography angiography and rotational digital subtraction angiography (DSA). The volumes were imported into a Vitrea 3D workstation (Vital Images Inc.) and the vascular lumen of various vessels and pathologies were segmented using a "marching cubes" algorithm. The results were exported as Stereo Lithographic (STL) files and were further processed by smoothing, trimming, and wall extrusion (to add a custom wall to the model). The models were printed using a Polyjet printer, Eden 260V (Objet-Stratasys). To verify the phantom geometry accuracy, the phantom was reimaged using rotational DSA, and the new data was compared with the initial patient data. The most challenging part of the phantom manufacturing was removal of support material. This aspect could be a serious hurdle in building very tortuous phantoms or small vessels. The accuracy of the printed models was very good: distance analysis showed average differences of 120 μm between the patient and the phantom reconstructed volume dimensions. Most errors were due to residual support material left in the lumen of the phantom. Despite the post-printing challenges experienced during the support cleaning, this technology could be a tremendous benefit to medical research such as in device development and testing.


title: Representations of swine flu: perspectives from a Malaysian pig farm.

text: Novel influenza viruses are seen, internationally, as posing considerable health challenges, but public responses to such viruses are often rooted in cultural representations of disease and risk. However, little research has been conducted in locations associated with the origin of a pandemic. We examined representations and risk perceptions associated with swine flu amongst 120 Malaysian pig farmers. Thirty-seven per cent of respondents felt at particular risk of infection, two-thirds were somewhat or very concerned about being infected. Those respondents who were the most anxious believed particular societal "out-groups" (homosexuals, the homeless and prostitutes) to be at higher infection risk. Although few (4%) reported direct discrimination, 46% claimed friends had avoided them since the swine flu outbreak. Findings are discussed in the context of evolutionary, social representations and terror management theories of response to pandemic threat.


title: Fasciocutaneous Lotus Petal Flap for Perineal Wound Reconstruction after Extralevator Abdominoperineal Excision: Application for Reconstruction of the Pelvic Floor and Creation of a Neovagina.

text: BACKGROUND The extralevator abdominoperineal excision (ELAPE) procedure creates an extensive soft tissue defect of the pelvic floor. It has been suggested that primary reconstruction reduces the risk of wound infection and delayed wound healing in this high-risk area. Use of myocutaneous flaps or omentoplasty are associated with functional limitations and complications. We performed the perineal variant of the lotus petal flap, which was originally described for vulvar reconstruction. We aimed to verify if application of the lotus petal flap in pelvic floor reconstruction after ELAPE meets the goals of an ideal reconstruction. METHODS We performed a retrospective study of 28 patients who underwent the lotus petal flap procedure for pelvic floor reconstruction after ELAPE between January 2011 and March 2014. RESULTS Median age was 62.1 years and 78.6 % of patients were female. In most patients the tumor was preoperatively irradiated (89.3 %) and in 28.6 % of the reconstructions a biological mesh was applied. No total flap loss occurred. Six (21.4 %) patients had no complications, while 13 (46.4 %) patients had minor complications (Clavien-Dindo grade I-II). Reoperation (Clavien-Dindo grade IIIb) was performed in nine patients (32.1 %), three of whom required a second lotus petal flap reconstruction. Median time until wound healing was 14 weeks. No additional surgery was performed for aesthetic problems. CONCLUSIONS Reconstruction of the pelvic floor after ELAPE using the fasciocutaneous lotus petal flap has limited major complications, but still with a high incidence of minor wound complications. This retrospective cohort study shows limited consequences on form and function.


title: A case of ruptured intrameatal aneurysm successfully treated with coil embolization.

text: Aneurysms within the internal acoustic canal are rare. We report the case of a 71-year-old female with subarachnoid hemorrhage resulting from a ruptured distal anterior inferior cerebellar artery which was not detected on initial radiological examination. A second rupture was detected by contrast-enhanced computed tomography and successfully treated by endovascular coil embolization. The patient recovered without neurological deficits. To the best of our knowledge, this is the first report of an intrameatal aneurysm treated by endovascular coil embolization. We suggest endovascular coil embolization as an alternative to open surgery, even in cases of deep intrameatal aneurysm.


title: Sensitivity and uncertainty analyses for a sars model with time-varying inputs and outputs.

text: This paper presents a statistical study of a deterministic model for the transmission dynamics and control of severe acute respiratory syndrome (SARS). The effect of the model parameters on the dynamics of the disease is analyzed using sensitivity and uncertainty analyses. The response (or output) of interest is the control reproduction number, which is an epidemiological threshold governing the persistence or elimination of SARS in a given population. The compartmental model includes parameters associated with control measures such as quarantine and isolation of asymptomatic and symptomatic individuals. One feature of our analysis is the incorporation of time-dependent functions into the model to reflect the progressive refinement of these SARS control measures over time. Consequently, the model contains continuous time-varying inputs and outputs. In this setting, sensitivity and uncertainty analytical techniques are used in order to analyze the impact of the uncertainty in the parameter estimates on the results obtained and to determine which parameters have the largest impact on driving the disease dynamics.


New Books.


Neonatal enterocolitis associated with coronavirus infection in a foal: a case report.


title: Rasch Analysis of The WURSS-21 Dimensional Validation and Assessment of Invariance.

text: BACKGROUND The purpose of this study is to use Rasch analysis to explore the validity of considering self-report scores from Wisconsin Upper Respiratory Symptom Survey (WURSS-21) as a single global illness severity domain. The WURSS-21 is a widely used questionnaire instrument that assesses symptom severity and functional impact of common cold and flu-like illness. METHODS This study applies item response theory, specifically Rasch modeling, to investigate dimensional and measurement properties of the WURSS-21, and looks at invariance over time. The data assessed represents 1167 people, each scoring the WURSS-21 once daily for up to seven consecutive days of acute upper respiratory infection (URI) illness. RESULTS Rasch analysis supports a single domain WURSS-21 global symptom score. Assessment of differential item functioning across seven days of illness provides evidence for measurement invariance. While individual items rating physical symptoms were somewhat variable, items rating functional impairment and quality of life impact appeared quite consistent across a single domain over seven days of illness. CONCLUSION Rasch analysis of WURSS-21 items provides evidential support for a single invariant domain. These findings support the practice of using a simply summed daily global illness severity score to represent the overall symptomatic and functional impairments arising from URI.


Health care in Canada.


title: Kinetics of serological responses in influenza A(H7N9)-infected patients correlate with clinical outcome in China, 2013.

text: The novel avian influenza A(H7N9) infection has recently emerged to cause severe respiratory illness in China. The objectives of this study were to define the kinetics of the antibody responses in patients with influenza A(H7N9) disease and to correlate these kinetics with clinical outcome. Serial serum samples were obtained at intervals of three to four days from 18 patients with virologically confirmed A(H7N9) disease in Shanghai. We determined the kinetics of the haemagglutination inhibition (HI) and A(H7H9) pseudotype neutralisation antibody (Nab) responses and correlated these with clinical outcomes. Most patients had robust serological responses by both HI and Nab tests. Taking into account censoring due to time of testing and death, the median time from onset of illness to Nab titre ≥1:40 was 14 days (95% confidence interval (CI): 11–18 days) in the fatal cases and 10.5 days (95% CI: 7–12) in the survivors (p=0.003). The two groups did not differ in initial Nab titres, but the rate of increase in Nab titres was significantly faster for survivors by approximately 10-fold per 15 days (p=0.007). Early and rapid induction of Nab was correlated significantly with better clinical outcome.


title: Seroepidemiology of Middle East respiratory syndrome (MERS) coronavirus in Saudi Arabia (1993) and Australia (2014) and characterisation of assay specificity.

text: The pseudoparticle virus neutralisation test (ppNT) and a conventional microneutralisation (MN) assay are specific for detecting antibodies to Middle East respiratory syndrome coronavirus (MERS-CoV) when used in seroepidemiological studies in animals. Genetically diverse MERS-CoV appear antigenically similar in MN tests. We confirm that MERS-CoV was circulating in dromedaries in Saudi Arabia in 1993. Preliminary data suggest that feral Australian dromedaries may be free of MERS-CoV but larger confirmatory studies are needed.


title: Potential for antiviral treatment of severe acute respiratory syndrome.

text: PURPOSE OF REVIEW Severe acute respiratory syndrome is a new, sometimes lethal disease of humans that is caused by a novel coronavirus. To date there have been over 750 related deaths and there is clearly an urgent need to develop specific antiviral drugs to combat this disease. In this review, the authors shall focus on the molecular biology of the coronavirus and suggest how this information can be used to identify possible targets for antiviral drugs. RECENT FINDINGS Within a remarkably short period of time, the severe acute respiratory syndrome coronavirus has been isolated, its genome has been sequenced and the structure of at least one key viral enzyme has been deduced. In addition, bioinformatic analysis has predicted a number of enzymatic activities associated with proteins of the viral replicase-transcriptase complex. In some cases, these functions have been confirmed by biochemical analysis. Thus, there has been significant progress in the rational approach to anti-severe acute respiratory syndrome coronavirus drug design. This approach, combined with the random screening of licensed compounds or existing compound libraries, should result in the identification of novel lead compounds and the expeditious development of antiviral drugs. SUMMARY Although the initial severe acute respiratory syndrome epidemic has been controlled by conventional measures, the animal reservoir for the coronavirus progenitor has not been identified. It is therefore likely that the virus will be reintroduced into the human population in the future. When this happens, the most economical and effective way to contain the virus will be the therapeutic use of antiviral drugs.


title: Intestinal adenocarcinoma with pancreas and lymph node metastases in a captive cotton-top tamarin (Saguinus oedipus).

text: A case of intestinal adenocarcinoma with metastases to the pancreas and regional lymph node was found in a 9-year-old, captive female cotton-top tamarin (Saguinus oedipus) with intermittent diarrhea. At necropsy, the tumor mass was located in the ileo-cecal junction causing circumferential thickening and annular stenosis. Microscopically, the lesions at primary and metastatic sites showed typical features of mucinous adenocarcinoma as seen in humans, including intracellular and extracellular mucin production and characteristic appearance of a signet ring of the tumor cells. The diagnosis was confirmed by histological evaluation, positive cytokeratin immunostain, and mucin production demonstrated by PAS and Alcian blue stain. It is speculated that the development of intestinal carcinoma was partly attributable to the excessive absorption of a diet of refined food, unbalanced nutrition, and the nature of these animals to develop stress easily.


MSMA Recommends "Shelter-In-Place" to Missouri Governor to Curb COVID-19.


title: Incidence and sequelae of acute pelvic inflammatory disease among active component females, U.S. Armed Forces, 1996-2016.

text: Pelvic inflammatory disease (PID) is a consequence of untreated or inadequately treated sexually transmitted infections that can itself lead to infertility and ectopic pregnancy. Annual screening for asymptomatic chlamydia and gonorrhea infection helps reduce the incidence of acute PID. In the military, routine versus risk-based individual screening for chlamydia and gonorrhea began in some services in 2001, with full implementation across the services by 2005. From 1996 through 2016, rates of acute PID among active component women started to decline in the mid-2000s, consistent with national trends and coinciding with implementation of routine annual chlamydia and gonorrhea screening. Of active component women diagnosed with acute PID from 1996 through 2012, 6.1% were subsequently diagnosed with infertility or ectopic pregnancy, with higher proportions found among women in the Army and those aged 25-34 years. The overall decrease in the rates of acute PID in the military is consistent with national trends, and continued screening for asymptomatic chlamydia and gonorrhea infections should result in a continuing decline in the incidence of acute PID.


title: Etiology of diarrhea in measles.

text: Thirty-six measles cases with diarrhea were studied with two age and sex matched control groups, measles without diarrhea (75 cases) and acute diarrhea (70 cases). Bacterial pathogens were isolated from 5 out of 36 (13.9%) in measles with diarrhea but rotavirus, coronavirus and parasites were not detected in any case. The bacterial and viral etiology of measles with diarrhea were statistically significant different from the acute diarrhea group (p less than 0.005) and p less than 0.01 respectively) but not from the measles without diarrhea group (p greater than 0.05). 83.3 per cent of cases had diarrhea during 4 days before and after the appearance of rash. Watery diarrhea was frequently observed in this study (63.9%). Although measles virus was not identified in stools, data from this study suggested that measles may be the viral agent causing diarrhea. Watery stools are often observed and the presence of faecal white blood cell may be seen.


title: Effects of Sampling Conditions and Environmental Factors on Fecal Volatile Organic Compound Analysis by an Electronic Nose Device.

text: Prior to implementation of volatile organic compound (VOC) analysis in clinical practice, substantial challenges, including methodological, biological and analytical difficulties are faced. The aim of this study was to evaluate the influence of several sampling conditions and environmental factors on fecal VOC profiles, analyzed by an electronic nose (eNose). Effects of fecal sample mass, water content, duration of storage at room temperature, fecal sample temperature, number of freeze-thaw cycles and effect of sampling method (rectal swabs vs. fecal samples) on VOC profiles were assessed by analysis of totally 725 fecal samples by means of an eNose (Cyranose320®). Furthermore, fecal VOC profiles of totally 1285 fecal samples from 71 infants born at three different hospitals were compared to assess the influence of center of origin on VOC outcome. We observed that all analyzed variables significantly influenced fecal VOC composition. It was feasible to capture a VOC profile using rectal swabs, although this differed significantly from fecal VOC profiles of similar subjects. In addition, 1285 fecal VOC-profiles could significantly be discriminated based on center of birth. In conclusion, standardization of methodology is necessary before fecal VOC analysis can live up to its potential as diagnostic tool in clinical practice.


title: Journal impact factor in the era of expanding literature.

text: The Journal Citation Reports (JCR) of the Science Citation Index (SCI), published by the Institute of Scientific Information (ISI), provides a comprehensive database for analysis of journals. Recent use of JCR's journal impact factor for evaluation of journals and authors has provoked strongly mixed reactions among investigators. This paper examines the effects of the rapidly expanding literature on the impact factor over the past decade and examines the limitations of journal impact factor for evaluating individual author's contributions. The JCR analyzed 6088 journals in 2005, a 32% increase in new listings since 1995. During the same period, there was a 39% increase in new journal listings in the infectious diseases category. The phenomenon of journal proliferation has had a profound effect on the journal impact factor. During the past decade an increased impact factor was observed in 92% of the top ranking major journals, especially in young and rapidly expanding research fields. Certain highly cited new journals published primarily review articles - not original contributions. There was no increase in impact factor among some of the best known journals, such as Journal of Experimental Medicine, Proceedings of the National Academy of Sciences (USA), and Journal of Infectious Diseases. Clearly, journal impact factor is an imperfect tool for measuring the quality of articles, and its use in evaluating authors has inherent risks. In spite of its limitations, journal impact factor can be used as a rough indicator of scientific quality in specific subject categories and for serious reading and learning.


title: [Topical problems in the provision of the Russian Federation's population with sanitary-and-epidemiological well-being].

text: The most important political and organizational events considerably affecting the strategy and tactics of prevention of infectious diseases may be called at the Summit of Eight (Saint Petersburg) that has considered the problem of consolidation of the world community's efforts in counteracting the prevalence of infectious diseases, as well as the adopted specific decisions on this matter; development of a concept of financial provision of Russian initiatives in the control of infectious diseases and its realization by the Government of the Russian Federation; implementation of the national priority health project on the additional mass immunization against rubella, influenza, poliomyelitis, prevention and treatment of HIV/AIDS and viral hepatitis B and C; implementation of regional and federal programs on the population's sanitary-and-epidemiological well-being; further improvement and development of a legal and standard basis in the control of infectious diseases; reorganization of a state sanitary-and-epidemiological service on a large scale; conferment of the status of a poliomyelitis-free country and a WHO certificate to the Russian Federation; development and implementation of the national action plan to maintain the achieved results; implementation of the national program in eradicating measles in the Russian Federation; implementation of complex measures against the importation and spread of revived particularly dangerous infections, such as severe acute respiratory syndrome and avian influenza and preparation of the population for a possible influenza pandemic; establishment of an executive headquarters to coordinate activities against the prevalence of avian influenza in the Russian Federation; making decisions, with the results of a meeting of the Russian Federation's State Council being kept in mind, on the intensification and coordination of activities aimed at counteracting the epidemic of HIV/AIDS in the Russian Federation, by setting up a governmental commission; involvement in the solution of biological safety problems in accordance with the "Foundations of public chemical and biological safety policy of the Russian Federation for the period till 2000 and further prospects" approved by the President of the Russian Federation; implementation of measures for sanitary area control within the framework of the goal-oriented federal program "The State Boundary of the Russian Federation" (2003-2010).


title: Sequence comparison of the 5' end of mRNA 3 from transmissible gastroenteritis virus and porcine respiratory coronavirus.

text: Analysis of porcine transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV) mRNA species indicated a deletion in mRNA 3 of PRCV. Polymerase chain reaction (PCR) was used to clone the 5' end of mRNA 3 from PRCV for comparison with the equivalent region in TGEV. Small deletions were observed within and around the PRCV sequence equivalent to the putative open reading frame (ORF) ORF-3a identified in TGEV. The potential RNA polymerase-leader complex binding site (leader RNA binding site), ACTAAAC, found upstream of ORF-3a in TGEV, was absent from the PRCV genome but a potential site was found in the PRCV genome upstream of a gene equivalent to TGEV ORF-3b. PCR analysis, using primers corresponding to sequences within the ORF-3b gene and the leader RNA sequence, confirmed that the leader RNA binding site was upstream of a gene equivalent to TGEV ORF-3b on PRCV mRNA 3 but upstream of ORF-3a on TGEV mRNA 3. The presence of the new leader RNA binding site would be responsible for generating the smaller mRNA 3 species found in PRCV-infected cells.


title: [Standardized management guideline for pediatric wards of hematology and oncology during the epidemic of coronavirus disease 2019].

text: With the spread of coronavirus disease 2019 (COVID-19) and growing knowledge of its diagnosis and treatment, it has been clear that children are also susceptible to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The children with hematological tumors are a special population with immunosuppression and special therapeutic characteristics. Here the management guideline for pediatric wards of hematology and oncology during COVID-19 epidemic is established based on the features of children with hematological tumors.


title: The kinetics-based enzyme-linked immunosorbent assay for coronavirus antibodies in cats: calibration to the indirect immunofluorescence assay and computerized standardization of results through normalization to control values.

text: The computer-assisted, kinetics-based enzyme-linked immunosorbent assay for coronavirus antibodies in cats was calibrated to the conventional indirect immunofluorescence assay by linear regression analysis and computerized interpolation (generation of "immunofluorescence assay-equivalent" titers). Procedures were developed for normalization and standardization of kinetics-based enzyme-linked immunosorbent assay results through incorporation of five different control sera of predetermined ("expected") titer in daily runs. When used with such sera and with computer assistance, the kinetics-based enzyme-linked immunosorbent assay minimized both within-run and between-run variability while allowing also for efficient data reduction and statistical analysis and reporting of results.


title: Alternative roles for interferon-gamma in the immune response to DNA vaccines encoding related melanosomal antigens.

text: Tyrosinase-related proteins-1 and -2 (gp75/TRP-1 and TRP-2) are melanosomal membrane glycoproteins recognized by antibodies and T-cells from patients with melanoma. Xenogeneic DNA immunization against gp75/TRP-1 generates antibody-dependent tumor immunity and autoimmune depigmentation. In contrast xenogeneic TRP-2 DNA immunization induces immunity mediated by CD8+ T-cells. The role of IFN-gamma in the generation of tumor immunity and autoimmune depigmentation in these two models was investigated. No tumor protection and minimal depigmentation was observed after immunization with human TRP-2 DNA in mice deficient in IFN-gamma ligand. Repletion with recombinant murine IFN-gamma restored tumor immunity. Experiments using IL4 deficient mice demonstrated that tumor immunity was unaffected but that autoimmune depigmentation was potentially accelerated, consistent with down-modulation of autoimmunity against TRP-2 by IL4. In contrast, IFN-gamma was not required for the generation of immunity to gp75/TRP-1. In fact, exogenous IFN-gamma ablated autoantibody responses against gp75/TRP-1 after xenogeneic DNA immunization, consistent with a down-regulatory effect of IFN-gamma. These results show that immunity to TRP-2 following DNA immunization uses an IFN-gamma-dependent Th1 pathway, but immunity to gp75/TRP-1 is down-regulated by IFN-gamma.


Acceptability and technical problems of the female condom in Malawi.


title: Comparison of virus isolation, immunofluorescence and electron microscopy for the diagnosis of animal viruses.

text: Results of 539 virus-positive samples from animals during the past three years were analyzed to determine the sensitivity of three different techniques i.e. virus isolation (VI), direct immunofluorescence (DFA), and electron microscopy (EM). The results showed that most of the positive cases were diagnosed with the help of all three techniques, either alone or in different combinations. While VI and DFA were more effective for the detection of bovine viral diarrhea, pseudorabies and infectious bovine rhinotracheitis viruses, EM was more sensitive for the detection of rota- and coronaviruses. The need for selective use of these tests for rapid diagnosis and the role of different factors affecting the sensitivity of these tests are discussed.


title: Isolation of avian infectious bronchitis coronavirus from domestic peafowl (Pavo cristatus) and teal (Anas).

text: Coronavirus-like viruses, designated peafowl/China/LKQ3/2003 (pf/CH/LKQ3/03) and teal/China/LDT3/2003 (tl/CH/LDT3/03), were isolated from a peafowl and a teal during virological surveillance in Guangdong province, China. Partial genomic sequence analysis showed that these isolates had the S-3-M-5-N gene order that is typical of avian coronaviruses. The spike, membrane and nucleocapsid protein genes of pf/CH/LKQ3/03 had >99 % identity to those of the avian infectious bronchitis coronavirus H120 vaccine strain (Massachusetts serotype) and other Massachusetts serotype isolates. Furthermore, when pf/CH/LKQ3/03 was inoculated experimentally into chickens (specific-pathogen-free), no disease signs were apparent. tl/CH/LDT3/03 had a spike protein gene with 95 % identity to that of a Chinese infectious bronchitis virus (IBV) isolate, although more extensive sequencing revealed the possibility that this strain may have undergone recombination. When inoculated into chickens, tl/CH/LDT3/03 resulted in the death of birds from nephritis. Taken together, this information suggests that pf/CH/LKQ3/03 might be a revertant, attenuated vaccine IBV strain, whereas tl/CH/LDT3/03 is a nephropathogenic field IBV strain, generated through recombination. The replication and non-pathogenic nature of IBV in domestic peafowl and teal under field conditions raises questions as to the role of these hosts as carriers of IBV and the potential that they may have to transmit virus to susceptible chicken populations.


title: Mechanism of stimulation of plus-strand synthesis by an RNA replication enhancer in a tombusvirus.

text: Replication of RNA viruses is regulated by cis-acting RNA elements, including promoters, replication silencers, and replication enhancers (REN). To dissect the function of an REN element involved in plus-strand RNA synthesis, we developed an in vitro trans-replication assay for tombusviruses, which are small plus-strand RNA viruses. In this assay, two RNA strands were tethered together via short complementary regions with the REN present in the nontemplate RNA, whereas the promoter was located in the template RNA. We found that the template activity of the tombusvirus replicase preparation was stimulated in trans by the REN, suggesting that the REN is a functional enhancer when located in the vicinity of the promoter. In addition, this study revealed that the REN has dual function during RNA synthesis. (i) It binds to the viral replicase. (ii) It interacts with the core plus-strand initiation promoter via a long-distance RNA-RNA interaction, which leads to stimulation of initiation of plus-strand RNA synthesis by the replicase in vitro. We also observed that this RNA-RNA interaction increased the in vivo accumulation and competitiveness of defective interfering RNA, a model template. We propose that REN is important for asymmetrical viral RNA replication that leads to more abundant plus-strand RNA progeny than the minus-strand intermediate, a hallmark of replication of plus-strand RNA viruses.


title: Treatment of severe acute respiratory syndrome with convalescent plasma.

text: In March 2003, an outbreak of severe acute respiratory syndrome started in Hong Kong. A 57-year-old woman had a typical presentation, including fever, non-productive cough, malaise, lymphopenia, and raised liver aminotransferases. The clinical course and successful treatment with convalescent plasma, ribavirin, and corticosteroids are discussed.