title: Fgl2 deficiency causes neonatal death and cardiac dysfunction during embryonic and postnatal development in mice.

text: We hypothesized that cardiac dysfunction was responsible for the high perinatal lethality that we previously reported in fibrinogen-like protein 2 (Fgl2) knockout (KO) mice. We therefore used ultrasound biomicroscopy to assess left ventricular (LV) cardiac structure and function during development in Fgl2 KO and wild-type (WT) mice. The only deaths observed between embryonic day (E)8.5 (onset of heart beating) and postnatal day (P)28 (weaning) were within 3 days after birth, when 33% of Fgl2 KO pups died. Histopathology and Doppler assessments suggested that death was due to acute congestive cardiac failure without evidence of valvular or other obvious cardiac structural abnormalities. Heart rates in Fgl2 KO embryos were significantly reduced at E8.5 and E17.5, and irregular heart rhythms were significantly more common in Fgl2 KO (21/26) than WT (2/21) embryos at E13.5. Indexes of systolic and/or diastolic cardiac function were also abnormal in KO mice at E13.5 and E17.5, in postnatal mice studied at P1, and in KO mice surviving to P28. M-mode analysis showed no difference in LV diastolic chamber dimension, although posterior wall thickness was thinner at P7 and P28 in Fgl2 KO mice. We conclude that Fgl2 deficiency is not associated with obvious structural cardiac defects but is associated with a high incidence of neonatal death as well as contractile dysfunction and rhythm abnormalities during embryonic and postnatal development in mice.


title: Structure and intracellular assembly of the transmissible gastroenteritis coronavirus.

text: Coronaviruses have been described as pleomorphic, round particles with a helical nucleocapsid as the unique internal structure under the virion envelope. Our studies on the organization of the transmissible gastroenteritis coronavirus (TGEV) have shown that the structure of these viruses is more complex. Different electron microscopy techniques, including cryomicroscopy of vitrified viruses, revealed the existence of an internal core, most probably icosahedral, in TGEV virions. Disruption of these cores induced the release of elongated ribonucleoprotein complexes. Ultrastructural analysis of freeze-substituted TGEV-infected swine testis (ST) cells showed characteristic intracellular budding profiles as well as two types of virions. While large virions with an electron-dense internal periphery are seen at perinuclear regions, smaller viral particles exhibiting compact internal cores of poligonal contours are more abundant in areas closer to the plasma membrane of the cell. These data strongly suggest that maturation events following the budding process are responsible for the formation of the internal core shell, the new structural element that we have recently described in extracellular infectious TGEV virions.


title: Severe acute respiratory syndrome: overview with an emphasis on the Toronto experience.

text: OBJECTIVE To provide an overview of the severe acute respiratory syndrome (SARS) outbreak in Toronto, Ontario, which experienced the largest outbreak outside Asia, and to review what has been learned during the past year. DATA SOURCES MEDLINE search of all studies related to SARS, including review of the Centers for Disease Control and Prevention, World Health Organization (WHO), and Health Canada Web sites. DATA SYNTHESIS During the SARS outbreak in Toronto, 438 people had been diagnosed as having suspected or probable SARS and 44 people died. Elderly people and those with comorbid illnesses were at greatest risk of complications or death. Transmission was via direct contact with respiratory secretions. The use of gloves, gowns, N95 masks, and eye protection was effective in preventing transmission. No transmission occurred before symptom onset or after recovery. Serologic tests suggest that antibodies may not appear until 28 days after illness onset. Molecular tests give their greatest yield during the second week of illness. The value of ribavirin treatment remains questionable. The combination of interferon plus corticosteroids appears to be better than corticosteroids alone. Postmortem examination revealed pulmonary edema and evidence of diffuse alveolar damage. Very few morphological changes were noted in other organs despite the presence of viral RNA as detected by polymerase chain reaction. CONCLUSION On July 5, 2003, the WHO declared that the SARS outbreak was over. Since then, new cases of SARS have been reported in Asia. With global travel, the disease can rapidly spread throughout the world. Therefore, we must remain vigilant to prevent another pandemic.


docid: kbwnhqmw

title: COVID-19: Striking a Balance between Health Care Provider Safety and Patient Care in the Indian Hospital Settings.

text: Background The COVID-19 pandemic has taken its toll on the health care systems all over the world. The global shortage of resources has adversely affected the safety of healthcare personnel as well patients. This has forced the health care facilities to adopt various measures and improvisation in infection control practices, in addition to those in personnel protective equipments (PPEs) adapting to their need and their resources. Objectives To review the literature on the infection control practices in health care facilities against COVID-19 and put forward practical solutions, in addition to personnel protective equipments that may be adopted for health care personnel and patient safety in the Indian setting. Conclusions We are now amidst of COVID-19 pandemic and we need to focus on 'Risk reduction as much as possible'. The facilities and resources should transform to meet the challenges within the shortest time frame. The concept of patient care needs to balance with the safety of the healthcare, patient and society. This is achievable only by a combination of engineering controls, administrative controls and PPEs. It involves a lot of committed team work while managing an outbreak like COVID - 19 attached with a lot of social stigma.


title: Structures of proteases for ubiqutin and ubiquitin-like modifiers.

text: Post-translational modifiers can alter the function of proteins in many different ways. The conjugation of ubiquitin (Ub) and ubiqutin-like modifiers (Ubls) to proteins has been shown to be especially crucial in regulating a variety of cellular processes including the cell cycle, growth control, quality control, localization and many more. It is a highly dynamic process and involves a number of enzymes called E1, E2 and E3. Ub and Ubls are removed from the target proteins by deubiquitinating enzymes (DUBs) or Ubl-specific proteases (ULPs), thereby deconjugation can act as an additional level of control over the ubiquitin-conjugation system. In addition, DUBs and ULPs are responsible for activating Ub and Ubls from their inactive corresponding precursor forms. Here we review recent progress in molecular details of these deconjugating enzymes of Ubls.


title: Organizational culture influences health care workers' influenza immunization behavior.

text: BACKGROUND AND OBJECTIVES Low rates of influenza immunization among health care workers (HCWs) pose a potential health risk to patients in primary care practices. Despite previous educational efforts and programs to reduce financial barriers, HCW influenza immunization rates remain low. Variation in practice-level organizational culture may affect immunization rates. To explore this relationship, we examined organizational cultures and HCWs' influenza immunization behaviors in three family medicine practices. METHODS We used a multi-method comparative case study. A field researcher used participant observation, in-depth interviews, and key informant interviews to collect data in each practice in November-December 2003. A diverse team used grounded theory to analyze text data. RESULTS Organizational culture varied among practices and differing HCW immunization rates were observed. The most structured and business-like practice achieved immunization of all HCWs, while the other two practices exhibited greater variation in HCW immunization rates. Physicians in the practices characterized as chaotic/disorganized or divided were immunized at higher rates than other members of the practices. CONCLUSIONS In these practices, organizational culture was associated with varying rates of influenza immunization for HCWs, especially among nonphysicians. Addressing elements of organizational culture such as beliefs regarding influenza immunization and office policies may facilitate the immunization of all staff members.


title: The ethics of responding to a novel pandemic.

text: Recent epidemics and pandemics have highlighted a number of ethical concerns about the response to the increasing threat of emerging infectious diseases. Some of these ethical concerns are very fundamental. They include why a pandemic was declared, how much clinical information can be collected for public health without threatening patient confidentiality and how to ensure fairness in the distribution of resources. We discuss these issues and suggest approaches to resolve these dilemmas as we anticipate the next pandemic.


title: Comparison of targeting F and G protein genes to detect bovine and ovine respiratory syncytial viruses.

text: In this study, 2 reverse transcriptase-polymerase chain reaction (RT-PCR) assays were developed and compared for simultaneous detection of bovine and ovine respiratory syncytial viruses (RSVs). One assay was based on a set of primers, which amplified a 426-bp fragment of either bovine or ovine RSV F gene (RT-PCR F). The F products could be distinguished by EcoRI or BstYI restriction endonuclease cleavage. In the other assay, a set of primers amplified a 542-bp fragment of either ovine or bovine RSV G gene (RT-PCR G). EcoO1091 and RsaI restriction enzymes were used to differentiate between the ovine and bovine PCR-G products. Sequencing of the PCR products confirmed the fidelity of both assays. The 2 assays were evaluated using 18 bovine RSV isolates, 1 ovine RSV, 1 bighorn sheep RSV isolate, 1 caprine RSV isolate, 2 human RSV isolates, and several other viruses associated with bovine respiratory tract disease. RT-PCR G may be more sensitive in detecting viral RNA. Because the target sequence of the F gene is more conserved than that of the G gene, RT-PCR F followed by the appropriate restriction enzyme cleavage may be superior to RT-PCR G to discriminate between the 2 ruminant RSV subgroups. This assay should prove useful for determining the relative contribution of ovine and bovine RSV to the pathogenesis of bovine respiratory tract disease.


title: Coronavirus and Pasteurella infections in bovine shipping fever pneumonia and Evans' criteria for causation.

text: Respiratory tract infections with viruses and Pasteurella spp. were determined sequentially among 26 cattle that died during two severe epizootics of shipping fever pneumonia. Nasal swab and serum samples were collected prior to onset of the epizootics, during disease progression, and after death, when necropsies were performed and lung samples were collected. Eighteen normal control cattle also were sampled at the beginning of the epizootics as well as at weekly intervals for 4 weeks. Respiratory bovine coronaviruses (RBCV) were isolated from nasal secretions of 21 and 25 cattle before and after transport. Two and 17 cattle nasally shed Pasteurella spp. before and after transport, respectively. RBCV were isolated at titers of 1 x 10(3) to 1.2 x 10(7) PFU per g of lung tissue from 18 cattle that died within 7 days of the epizootics, but not from the lungs of the remaining cattle that died on days 9 to 36. Twenty-five of the 26 lung samples were positive for Pasteurella spp., and their CFU ranged between 4.0 x 10(5) and 2.3 x 10(9) per g. Acute and subacute exudative, necrotizing lobar pneumonia characterized the lung lesions of these cattle with a majority of pneumonic lung lobes exhibiting fibronecrotic and exudative changes typical of pneumonic pasteurellosis, but other lung lobules had histological changes consisting of bronchiolitis and alveolitis typical of virus-induced changes. These cattle were immunologically naive to both infectious agents at the onset of the epizootics, but those that died after day 7 had rising antibody titers against RBCV and Pasteurella haemolytica. In contrast, the 18 clinically normal and RBCV isolation-negative cattle had high hemagglutinin inhibition antibody titers to RBCV from the beginning, while their antibody responses to P. haemolytica antigens were delayed. Evans' criteria for causation were applied to our findings because of the multifactorial nature of shipping fever pneumonia. This analysis identified RBCV as the primary inciting cause in these two epizootics. These viruses were previously not recognized as a causative agent in this complex respiratory tract disease of cattle.


title: Coronaviruses as vectors: position dependence of foreign gene expression.

text: Coronaviruses are the enveloped, positive-stranded RNA viruses with the largest RNA genomes known. Several features make these viruses attractive as vaccine and therapeutic vectors: (i) deletion of their nonessential genes is strongly attenuating; (ii) the genetic space thus created allows insertion of foreign information; and (iii) their tropism can be modified by manipulation of the viral spike. We studied here their ability to serve as expression vectors by inserting two different foreign genes and evaluating systematically the genomic position dependence of their expression, using a murine coronavirus as a model. Renilla and firefly luciferase expression cassettes, each provided with viral transcription regulatory sequences (TRSs), were inserted at several genomic positions, both independently in different viruses and combined within one viral genome. Recombinant viruses were generated by using a convenient method based on targeted recombination and host cell switching. In all cases high expression levels of the foreign genes were observed without severe effects on viral replication in vitro. The expression of the inserted gene appeared to be dependent on its genomic position, as well as on the identity of the gene. Expression levels increased when the luciferase gene was inserted closer to the 3' end of the genome. The foreign gene insertions generally reduced the expression of upstream viral genes. The results are consistent with coronavirus transcription models in which the transcription from upstream TRSs is attenuated by downstream TRSs. Altogether, our observations clearly demonstrate the potential of coronaviruses as (multivalent) expression vectors.


title: Renin-angiotensin system, hypertension, and chronic kidney disease: pharmacogenetic implications.

text: About 80% of CKD (chronic kidney disease) patients are hypertensive, and kidney function and blood pressure are clearly related to both physiologic and pathologic conditions in a "vicious cycle". In this pathologic scenario, there is a renin-angiotensin system (RAS) hyperactivity associated to progression of renal damage. Current guidelines indicate as the first choice of antihypertensive intervention, the pharmacologic blockade of the RAS. Nonetheless, both response to treatment and renal protection have considerable inter-individual variability. The main aims of this review are to describe the genetic characteristics of RAS components and to identify the possible pharmacogenetic implications for RAS-blocker drugs in the hypertension-CKD scenario. To date, RAS polymorphisms have not been consistently associated to antihypertensive response and studies focusing on CKD are scarce. Nonetheless, pharmacogenetic studies for the RAS-blocker drugs could still be further explored, especially with new generation tools and focusing not only on the antihypertensive response, but also on renal protection as well.


Severe acute respiratory syndrome (SARS)--150 days on.


title: Laparoscopic and mini-invasive surgery.

text: BACKGROUND JGCA Gastric Cancer Treatment Guidelines (2004) include Laparoscopic Assisted Distal Gastrectomy (LADG) within the chapter of modified surgery. A metanalysis published in 2010 shows that LADG is significantly superior to Open Distal Gastrectomy (ODG) if comparing short term outcomes. Oncologic results prove to be comparable to ODG by one RCT and 2 retrospective studies. Little evidence is available on Laparoscopic Total Gastrectomy and concerns are raised about long-term oncologic outcomes. MATERIALS AND METHODS Laparoscopic Subtotal Gastrectomy is carried out with 4 to 5 ports in the periumbilical region (Hasson trocar for laparoscope) and upper quadrants. After exploration of the abdominal cavity surgical steps include coloepiploic detachment, omentectomy, dissection of the gastrocolic ligament, division of the left gastroepiploic vessels, division of right gastroepiploic vessels, division of pyloric vessels. The duodenum is transected with a linear stapler. Incision of the lesser omentum and dissection of the hepatoduodenal ligament allows completion of D2 lymphadenectomy. The 4/5ths of the stomach are transected starting from the greater curve at the junction of left and right gastroepiploic arcades by linear stapler. Roux-en-Y loop reconstruction is performed through a stapled side-to-side gastro-jejunal anastomosis and a side-to-side jejuno-jejunal anastomosis. Reconstruction after Laparoscopic Total Gastrectomy is performed preferably by a side-to-side esophago-jejunal anastomosis according to Orringer. DISCUSSION AND CONCLUSION A robotic assisted approach adds precision on lymphadenectomy and reconstructive techniques.


title: Assuring public health professionals are prepared for the future: the UAB public health integrated core curriculum.

text: In response to calls to improve public health education and our own desire to provide a more relevant educational experience to our Master of Public Health students, the University of Alabama at Birmingham (UAB) School of Public Health designed, developed, and instituted a fully integrated public health core curriculum in the fall of 2001. This curriculum combines content from discipline-specific courses in biostatistics, environmental health, epidemiology, health administration, and the social and behavioral sciences, and delivers it in a 15 credit hour, team-taught course designed in modules covering such topics as tobacco, infectious diseases, and emergency preparedness. Weekly skills-building sessions increase student competence in data analysis and interpretation, communication, ethical decision-making, community-based interventions, and policy and program planning. Evaluations affirm that the integrated core is functioning as intended: as a means to provide critical content in the core disciplines in their applied context. As public health education continues to be debated, the UAB public health integrated core curriculum can serve as one model for providing quality instruction that is highly relevant to professional practice.


title: Regulation of transcription of coronaviruses.

text: To study factors involved in regulation of transcription of coronaviruses, we constructed defective interfering (DI) RNAs containing sg RNA promoters at multiple positions. Analysis of the amounts of sg DI RNA produced by these DIs resulted in the following observations: (i) a downstream promoter downregulates an upstream promoter; (ii) an upstream promoter has little or no effect on the activity of a downstream promoter. Our data suggest that attenuation of upstream promoter activities by downstream promoter sequences plays an important role in regulating the amounts of sg RNAs produced by coronaviruses. Our observations are in accordance with the models proposed by Konings et al. and Sawicki and Sawicki.


title: Severe acute respiratory syndrome: management and reconfiguration of a radiology department in an infectious disease situation.

text: Severe acute respiratory syndrome, or SARS, is a new infectious disease pandemic with important public health concerns. The high infectivity rate by means of droplet transmission places health care workers at substantial risk of contracting the disease. Radiology departments are particularly affected, since imaging plays a vital role in both diagnosis and follow-up of this disease. The authors outline their experience in infection control and isolation procedures during this outbreak. Barrier precautions, reconfiguration of the department, separation of imaging equipment, cleaning procedures, personal protective equipment, and staff safety are discussed.


Use of defective RNAs containing reporter genes to investigate targeted recombination for avian infectious bronchitis virus.


title: Evidence for emergence of an amphibian iridoviral disease because of human-enhanced spread.

text: Our understanding of origins and spread of emerging infectious diseases has increased dramatically because of recent applications of phylogenetic theory. Iridoviruses are emerging pathogens that cause global amphibian epizootics, including tiger salamander (Ambystoma tigrinum) die-offs throughout western North America. To explain phylogeographical relationships and potential causes for emergence of western North American salamander iridovirus strains, we sequenced major capsid protein and DNA methyltransferase genes, as well as two noncoding regions from 18 geographically widespread isolates. Phylogenetic analyses of sequence data from the capsid protein gene showed shallow genetic divergence (< 1%) among salamander iridovirus strains and monophyly relative to available fish, reptile, and other amphibian iridovirus strains from the genus Ranavirus, suggesting a single introduction and radiation. Analysis of capsid protein sequences also provided support for a closer relationship of tiger salamander virus strains to those isolated from sport fish (e.g. rainbow trout) than other amphibian isolates. Despite monophyly based on capsid protein sequences, there was low genetic divergence among all strains (< 1.1%) based on a supergene analysis of the capsid protein and the two noncoding regions. These analyses also showed polyphyly of strains from Arizona and Colorado, suggesting recent spread. Nested clade analyses indicated both range expansion and long-distance colonization in clades containing virus strains isolated from bait salamanders and the Indiana University axolotl (Ambystoma mexicanum) colony. Human enhancement of viral movement is a mechanism consistent with these results. These findings suggest North American salamander ranaviruses cause emerging disease, as evidenced by apparent recent spread over a broad geographical area.


docid: pabvmcvl

title: A contribution for the forensic odontologist's safety in the autopsy room.

text: The Covid 19 pandemic we are currently facing all over the world is undoubtly an emergency condition which exposes all workers to the risks of a serious infection. The personnel working in the autoptic room is at high risk for the possible contact with infected material from corpses, splashes and aerosols diffusion, and must be always prepared to stay safe and healthy. We offer to the forensic odontologists' community a short summary of some old and new hints on the treatment of dead bodies in the autopsy room during this pandemic emergency time.


title: Prospective study of the incidence, aetiology and outcome of adult lower respiratory tract illness in the community.

text: BACKGROUND Acute lower respiratory tract illness in previously well adults is usually labelled as acute bronchitis and treated with antibiotics without establishing the aetiology. Viral infection is thought to be the cause in most cases. We have investigated the incidence, aetiology, and outcome of this condition. METHODS Previously well adults from a stable suburban population consulting over one year with a lower respiratory tract illness were studied. For the first six months detailed investigations identified predetermined direct and indirect markers of infection. Evidence of infection was assessed in relation to presenting clinical features, indirect markers of infection, antibiotic use, and outcome. RESULTS Consultations were very common, particularly in younger women (70/1000 per year in previously well women aged 16-39 years), mainly in the winter months; 638 patients consulted, of whom 316 were investigated. Pathogens were identified in 173 (55%) cases: bacteria in 82 (Streptococcus pneumoniae 54, Haemophilus influenzae 31, Moraxella catarrhalis 7), atypical organisms in 75 (Chlamydia pneumoniae 55, Mycoplasma pneumoniae 23), and viruses in 61 (influenza 23). Seventy nine (24%) had indirect evidence of infection. Bacterial and atypical infection correlated with changes in the chest radiograph and high levels of C reactive protein but not with (a) the GP's clinical assessment of whether infection was present, (b) clinical features other than focal chest signs, and (c) outcome, whether or not appropriate antibiotics were prescribed. CONCLUSIONS Over 50% of patients have direct and/or indirect evidence of infection, most commonly bacterial and atypical pathogens, but the outcome is unrelated to the identified pathogens. Many patients improve without antibiotics and investigations do not help in the management of these patients. GPs can reassure patients of the causes and usual outcome of this self-limiting condition.


title: Nosocomial acquisition of methicillin-resistant Staphylococcus aureus during an outbreak of severe acute respiratory syndrome.

text: OBJECTIVE The four hospitals assessed in this study use active surveillance cultures for methicillin-resistant Staphylococcus aureus (MRSA) and contact precautions for MRSA-positive patients as part of routine infection control practices. The objective of this study was to determine whether nosocomial acquisition of MRSA decreased in these hospitals during an outbreak of severe acute respiratory syndrome (SARS) when barrier precautions were routinely used for all patients. DESIGN Retrospective cohort study. SETTING Three tertiary-care hospitals (a 1100-bed hospital; a 500-bed hospital; and an 823-bed hospital) and a 430-bed community hospital, each located in Toronto, Ontario, Canada. PATIENTS All admitted patients were included. RESULTS The nosocomial rate of MRSA in all four hospitals combined during the SARS outbreak (3.7 per 10,000 patient-days) was not significantly different from that before (4.7 per 10,000 patient-days) or after (3.4 per 10,000 patient-days) the outbreak (P = .30 and P = .76, respectively). The nosocomial rate of MRSA after the outbreak was significantly lower than that before the outbreak (P = .003). Inappropriate reuse of gloves and gowns and failure to wash hands between patients on non-SARS wards were observed during the outbreak. Increased attention was paid to infection control education following the outbreak. CONCLUSIONS Inappropriate reuse of gloves and gowns and failure to wash hands between patients may have contributed to transmission of MRSA during the SARS outbreak. Attention should be paid to training healthcare workers regarding the appropriate use of precautions as a means to protect themselves and patients.


title: In vitro and in vivo expression of foreign genes by transmissible gastroenteritis coronavirus-derived minigenomes.

text: A helper-dependent expression system based on transmissible gastroenteritis coronavirus (TGEV) has been developed using a minigenome of 3.9 kb (M39). Expression of the reporter gene beta-glucuronidase (GUS) (2-8 microg per 10(6) cells) and the porcine respiratory and reproductive syndrome virus (PRRSV) ORF5 (1-2 microg per 10(6) cells) has been shown using a TGEV-derived minigenome. GUS expression levels increased about eightfold with the m.o.i. and were maintained for more than eight passages in cell culture. Nevertheless, instability of the GUS and ORF5 subgenomic mRNAs was observed from passages five and four, respectively. About a quarter of the cells in culture expressing the helper virus also produced the reporter gene as determined by studying GUS mRNA production by in situ hybridization or immunodetection to visualize the protein synthesized. Expression of GUS was detected in the lungs, but not in the gut, of swine immunized with the virus vector. Around a quarter of lung cells showing replication of the helper virus were also positive for the reporter gene. Interestingly, strong humoral immune responses to both GUS and PRRSV ORF5 were induced in swine with this virus vector. The large cloning capacity and the tissue specificity of the TGEV-derived minigenomes suggest that these virus vectors are very promising for vaccine development.


Infection control in mass-casualty respiratory care: research needs and future directions.


title: [Novel Research Progress of the Recombinant Vaccine for MERS-CoV].

text: Respiratory syndrome coronavirus in the Middle East (MERS-Co V) has caused wide attention since it was discovered, and the design of effective vaccines for MERS-Co V becomes a hot area at present. Therefore, this review is aimed at novel research progress of the recombinant vaccine for MERS-Co V, including selection and improvement of animal model for vaccine test,construction and optimization of recombinant subunit vaccine, attempts at recombinant live vector vaccine based on varies of vectors, as well as the advantages of pseudovirus. In conclusion, we make a summary about the novel research progress of the recombinant vaccine for MERS-Co V and a promising prospect of its development in terms of both safety and effectiveness verification in the future will be demonstrated.


title: How social structures, space, and behaviors shape the spread of infectious diseases using chikungunya as a case study.

text: Whether an individual becomes infected in an infectious disease outbreak depends on many interconnected risk factors, which may relate to characteristics of the individual (e.g., age, sex), his or her close relatives (e.g., household members), or the wider community. Studies monitoring individuals in households or schools have helped elucidate the determinants of transmission in small social structures due to advances in statistical modeling; but such an approach has so far largely failed to consider individuals in the wider context they live in. Here, we used an outbreak of chikungunya in a rural community in Bangladesh as a case study to obtain a more comprehensive characterization of risk factors in disease spread. We developed Bayesian data augmentation approaches to account for uncertainty in the source of infection, recall uncertainty, and unobserved infection dates. We found that the probability of chikungunya transmission was 12% [95% credible interval (CI): 8-17%] between household members but dropped to 0.3% for those living 50 m away (95% CI: 0.2-0.5%). Overall, the mean transmission distance was 95 m (95% CI: 77-113 m). Females were 1.5 times more likely to become infected than males (95% CI: 1.2-1.8), which was virtually identical to the relative risk of being at home estimated from an independent human movement study in the country. Reported daily use of antimosquito coils had no detectable impact on transmission. This study shows how the complex interplay between the characteristics of an individual and his or her close and wider environment contributes to the shaping of infectious disease epidemics.


title: Acute respiratory distress syndrome.

text: Acute respiratory distress syndrome is the clinical manifestation of severe, acute lung injury. It is characterized by the acute onset of diffuse, bilateral pulmonary infiltrates secondary to noncardiogenic pulmonary edema, refractory hypoxia, and decreased lung compliance. Acute respiratory distress syndrome occurs most frequently in the setting of sepsis, aspiration of gastric contents, trauma, or multiple transfusions. Its complex pathophysiology involves an inciting local or systemic event that initiates pulmonary endothelial and epithelial damage and subsequent increased permeability. Tachypnea, hypoxia, and respiratory alkalosis are typical early clinical manifestations, and they are usually followed by the appearance of diffuse pulmonary infiltrates and respiratory failure within 48 hours. Early identification and treatment of the underlying disorder, along with aggressive supportive care, are essential. Experimental therapies, including those using nitric oxide and surfactant, have not been shown to improve mortality in patients with ARDS, but new therapeutic approaches such as low-volume ventilation have been shown to decrease mortality. Many patients who survive ARDS have permanent, mild to moderate impairment of lung function. Quality of life after hospitalization with ARDS may be poorer than that in similar patients without ARDS.


title: Infrared thermography to mass-screen suspected SARS patients with fever.

text: Fever greater than 38 degrees C is a cardinal sign of patients with the severe acute respiratory syndromes (SARS). To reduce the risk of nosocomial cross infections, screening all patients and visitors who visit hospitals and clinics for fever at the entrance of every hospital building has become a standard protocol in Taiwan during the SARS epidemic from mid-April to mid-June 2003. We used a digital infrared thermal imaging (DITI) system (Telesis Spectrum 9000 MB) to conduct mass screening of patients and visitors who entered the hospital to identify those with fever. The DITI system has two components: a sensor head and a PC imaging workstation. The sensor head is an optic-mechanical device which consists of imagining optics for focusing the infrared source information on the infrared detector. The infrared images are further converted into electrical signals, which are then processed for real-time display on the monitor. During the period from April 13 to May 12 2003, 72,327 outpatients and visitors entered Taipei Medical University-Wan Fang Hospital, Taipei, Taiwan. A total of 305 febrile patients (0.42%) was detected by infrared thermography. Among them, three probable SARS patients were identified after thorough studies including contact history, laboratory tests and radiology examinations. The findings suggests that infrared thermography was an effective and reliable tool ideal for mass-screening patients with fever in the initial phase of screening for SARS patients at a busy hospital which sees approximately 3,000 outpatients every weekday during the SARS epidemic.


Public health in Canada: what are the real issues?


title: PARENTS' AND HEALTHCARE PROVIDERS PERSPECTIVES ABOUT HOSPITALISED CHILDREN BEING VISITED BY OTHER CHILDREN IN NAIROBI, KENYA.

text: BACKGROUND Throughout history, visitation of the hospitalised child has always been restricted. The subject of whether hospitalised children should be visited by other children has been accorded minimal attention. OBJECTIVE To determine the current status of visitation of the hospitalised children and the parents and healthcare providers'views on hospitalised children being visited by other children. DESIGN A descriptive study carried out using quantitative and qualitative methods in two phases SETTING All paediatric wards at Kenyatta National Hospital and Gertrudes Children's Hospital. SUBJECTS Nurses, paediatricians and parents of hospitalised children. RESULTS A total of 161 parents participated in phase I of the study whereas 11 nurses not specialised in paediatric nursing, 13 paediatric nurses, 12 nurse managers, fur paediatricians and13 parents participated in phase II of the study. The study established that visiting of the hospitalised child by family members especially children aged below twelve years is severely restricted particularly in the public hospital. Despite this, however, majority of the healthcare providers and the parents acknowledged the importance of the hospitalised children being visited by other children. This is because it promotes healing, gives the sick child psychological satisfaction and relieves anxiety in the hospitalised child, the accompanying parent and the other children. The risk of exposing the visiting children to infection was cited as the main reason for the restrictions. CONCLUSION Both the healthcare providers and the hospitalised children's parents appreciate the importance of the hospitalised child being visited by other children. There is a need to review healthcare policies to make provision for hospitalised children to be visited by other children.


title: Intra-corporeal anastomosis in laparoscopic surgery for benign colorectal disease.

text: INTRODUCTION Recent studies have described the benefits of performing bowel resections for benign and malignant diseases as a total laparoscopic procedure with intra-corporeal anastomosis. We describe our preliminary experience with total laparoscopic surgery for benign lower gastrointestinal diseases. METHODS All patients (n = 32) who had received elective bowel surgery with intra-corporeal anastomosis for benign diseases between November 2013 and August 2015 were included in this retrospective study. RESULTS A total of 15 patients (47%) had a primary anastomosis, 17 patients (53%) had a stoma reversal. The median length of stay was five days (range: 3-16). Surgical complications requiring re-operation were present in five patients (Clavien-Dindo grade IIIb) of whom two presented with anastomotic leaks (6%; 95% confidence interval: 2-20%). No post-operative medical complications according to the Clavien-Dindo Classification were seen. The median duration of surgery was 127 min. (range: 61-211 min.). No deaths occurred. CONCLUSION A total laparoscopic approach to benign lower gastrointestinal diseases with intra-corporeal ana-stomosis was feasible. FUNDING none. TRIAL REGISTRATION The trial is registered with the Danish Data Protection Agency.


title: The urban geography of SARS: paradoxes and dilemmas in Toronto's health care.

text: BACKGROUND Severe acute respiratory syndrome (SARS) has impacted heavily on both Canada's society and its health care system. Quite unexpectedly, hospitals and health care workers became facilitators of disease diffusion. This experience has demonstrated the need for a comprehensive disease control strategy. AIMS The aims are threefold. First, to examine the subtle and changing spatial characteristics of SARS during the short but critical period of its rapid spread in the Greater Toronto Area. Second, to make salient three paradoxes, and their associated dilemmas, faced by nurses and other health care professionals challenged with caring for the sick and containing the rapid spread of the disease. Third, to propose some ways of approaching these dilemmas, as well as some broader preventative and mitigating strategies. METHODS The crosscutting concepts of 'clinical uncertainty' and 'flow of human contacts' are used to explain disease transmission characteristics and urban spatial diffusion and to guide the particular strategies developed. Evidence for the paper comes from public health records, governmental and non-governmental health statements and the initial epidemiological research on SARS. Direct insights are also gained from recent first-hand experiences of Toronto's health care system during the crisis. DISCUSSION The concepts of clinical uncertainty and the flow of human contacts provide in-depth insights that complement the findings of large-scale epidemiological studies, and help operationalize their general calls for enhanced control measures. The comprehensive disease control strategy proposed includes the creation of a hospital infrastructure specific to the containment of biological threats; an advisory coalition of disease control specialists; the development of a biological threat-and-containment simulation laboratory and three specific programmes in patient safety, risk assessment and community mobilization. CONCLUSION It is argued that containment alone, while a necessary and urgent priority, cannot be seen as an end in itself and might better be understood as one possible trajectory within a comprehensive problem-solving strategy. The experiences in Toronto may offer insights to other cities and countries that currently lack such strategies and hence may be vulnerable to similar outbreaks of dangerous infectious diseases.


Human metapneumovirus: a not so new virus.


title: [Change of T lymphocyte and its activated subsets in SARS patients].

text: OBJECTIVE To study abnormal changes of T lymphocyte and its activated subsets in severe acute respiratory syndrome (SARS) patients. METHODS Flow cytometer with multi-color flouroscence and hematology analyzer were used to detect the expression of T lymphocyte and its activated a subsets in 240 SARS patients including 50 cases of critical type and 190 cases of common type. RESULTS Statistical analysis by means of SAS software showed that there was significant decrease in absolute counts (AC) of T lymphocyte and its subsets in SARS patients when compared with normal people, while percentages (PC) of CD3+CD25+ and CD3+ HLA-DR+ subsets were increased markedly. Compared with common type, there was significant decrease in absolute counts of critical type of T lymphocyte, CD4+, CD25+CD3+, CD28+CD4+, and CD95+CD4+subsets. The ACs of T lymphocytes including CD4 and CD8 subsets in different phases were as below: III > II > I. The ACs of subsets involved in activation such as CD3+ HLA-DR+/lym, CD3+CD25+/lym, CD28+CD4+/CD4, CD28+CD8+/CD8, and CD38+CD4+/CD4 all were highest in group III. In addition, the AC and PC of CD95+CD4+/CD4 and CD95+CD8/CD8 subset in group III were highest while group I was lowest. CONCLUSIONS With depressing cellular immunity, the activation of T lymphocytes were suppressed obviously in SARS patients, especially for critical patients.


title: Viral hyperinfection of the central nervous system and high mortality after hematopoietic stem cell transplantation for treatment of Theiler's murine encephalomyelitis virus-induced demyelinating disease.

text: Theiler's murine encephalomyelitis virus (TMEV) establishes a persistent infection in the central nervous system (CNS) leading to an inflammatory demyelinating disease of the CNS in which the histology and clinical course is similar to multiple sclerosis (MS). Disease pathogenesis is primarily due to T-cell-mediated destruction of myelin, which has been attributed to cytopathic effects of the virus, but immune-mediated destruction of myelin mediated via both virus-specific and myelin-specific T cells appear to play the major role. To determine if bone marrow transplantation would be an effective therapy for a virus-initiated autoimmune disease and to better separate viral cytopathic effects from immune-mediated demyelination, we ablated the immune system of TMEV-infected animals with 1,100 cGy total body irradiation, and then the animal's immunity was reconstituted by transplantation of disease-susceptible SJL/J mice with syngeneic marrow or disease-susceptible DBA/2J with marrow from disease-resistant (C57Bl/6 x DBA/2)F1 (B6D2) donors. Hematopoietic transplant performed after onset of disease resulted in 42% mortality in SJL/J syngeneic transplants, 47% mortality in diseased DBA2 recipients restored with marrow from naive B6D2 donors, and 12% in diseased DBA2 recipients receiving marrow from B6D2 donors previously infected with TMEV. Delayed type hypersensitivity (DTH) to both virion and myelin proteins was decreased in surviving mice that underwent transplantation; however, CNS viral titers were significantly elevated compared with nontransplanted controls. We conclude that a functional immune system with appropriate T-cell responses are important in prevention of lethal cytopathic CNS effects from TMEV. Relevant to the clinical use of bone marrow transplantation, attempts to ablate the immune system in viral-mediated immune diseases or virus-initiated autoimmune disease may have acute and lethal consequences. Our results raise concern about the attempted use of autologous hematopoietic transplantation in patients with MS, an autoimmune disease with a suspected virus etiology, particularly if the graft is aggressively depleted of lymphocytes.


Globalization and the health of the health care workforce.


title: [Kawasaki disease].

text: Kawasaki disease is a systemic acute vasculitis of unknown etiology. It is the leading cause of acquired heart disease in children in the USA. It occurs more frequently in boys and eighty percent of the cases occur in children under five years of age. The disease rarely occurs after eight years and it can affect children of all races, with higher incidence among Asian descendants. Kawasaki disease is characterized by fever, bilateral non-exudative conjunctivitis, redness and swelling of the tongue, lips and oral mucosa, abnormalities in the extremities, cervical lymph node, and polymorphic exanthema. Aneurysms and stenoses of coronary arteries occur in approximately 20 to 25% of untreated patients and subsequently can lead to acute myocardial infarction and sudden death. Treatment with intravenous immunoglobulin is effective and should be initiated early to prevent cardiac sequel. The development of diagnostic tests, more specific treatment approaches and prevention of this potentially fatal disease in children depends on continuous advances in the determination of its pathogenesis.


title: The Tecumseh study of respiratory illness. VIII. Acute infection in chronic respiratory disease and comparison groups.

text: Individuals with chronic lung disease and their families were selected from the Tecumsch community along with similarly selected families as comparison groups and studied for 1-year periods. Occurence of acute respiratory illness was ascertained weekly by telephone and calculated as an annual rate. Persons with chronic bronchitis not only experienced more acute lower respiratory illness than healthy comparison subjects, but total illness rates were somewhat higher as well. Infection rates were determined from blood samples taken 3 times from each participant during the surveillance year. Antibody tests were performed for respiratory syncytial virus, para-influenza virus types 1, 2, and 3, influenza types A and B, coronavirus OC43, Mycoplasma pneumoniae, and Haemophilus influenzae. Differences in serologic infection rates among the subgroups of the population were similar to those seen in the clinical data, with more frequent infection among those with bronchitis than among the comparison subjects. This finding indicates that some degree of increased susceptibility to actual infection existed among those individuals with bronchitis. Influence of smoking on illness and infection rates was also examined. Infections were, in general, more frequent in smokers than in nonsmokers, but illness rates were reversed, suggesting that perception of disease differed in the 2 groups. Rates of illness and infection of other adults in the families of the index individuals with bronchitis were not influenced by the higher rates seen in the index individuals; however, it was of interest that children of persons with bronchitis did have somewhat higher rates of infection than children of comparison subjects.


title: Comparative analysis of Coronaviridae nucleocapsid and surface glycoprotein sequences.

text: We analyzed the nucleocapsid and surface proteins from several Coronaviridae viruses using an alignment-free computer program. Three isolates of novel, human coronavirus (SARS0CoV-2) (2019) that are responsible for the current pandemic and older SARS strains of human and animal coronaviruses were examined. The nucleocapsid and glycoprotein sequences are identical for the three novel 2019 human isolates and they are closely related to these sequences in six bat and human SARS coronaviruses. This strongly supports the bat origin of the pandemic, novel coronavirus. One surface glycoprotein fragment of 111 amino acids is the largest, conserved, common permutation in the examined bat SARS-like and human SARS viruses, including the Covid-19 virus. BLAST analysis confirmed that this fragment is conserved only in the human and bat SARS strains. This fragment likely is involved in infectivity and is of interest for vaccine development. Surface glycoprotein and nucleocapsid protein sequence homologies of 58.9% and 82.5%, respectively, between the novel SARS0CoV-2 strains and the human SARS (2018) virus suggest that existing anti-SARS vaccines may provide some protection against the novel coronavirus.


Correlation between MRI, computed tomographic findings and clinical signs in a case of ovine coenurosis.


title: Detection of feline coronavirus in captive Felidae in the USA.

text: Feline coronavirus (FCoV) is an important pathogen of domestic and nondomestic Felidae. Investigation into the prevalence of FCoV in exotic Felidae has relied primarily on serology. The usefulness of genetic detection of FCoV using reverse transcription and nested polymerase chain reaction (RT/nPCR) for viral screening was investigated. Seventy-five biologic samples, primarily feces, from captive felids from 11 institutions were tested using PCR. Serum samples collected from all but 12 of these animals were tested for antibodies to type I and type II FCoV by indirect immunofluorescence. Twenty-four animals were positive using RT/nPCR for virus. Twenty-nine animals were seropositive to type I and/or type II FCoV. From serologic data, infection with a virus antigenically related to FCoV type I occurred most commonly. Serology did not correlate with virus shedding because 13 animals were seronegative to FCoV type I and II but positive using RT/nPCR for virus. Conversely, 20 animals were seropositive but negative using RT/nPCR for FCoV. Some of the populations in which virus was detected had experienced health problems, including feline infectious peritonitis (FIP), necrotizing colitis, and mild enteritis. In addition to its role in FIP, this virus may play a role in gastrointestinal diseases of infected animals. This study demonstrates that FCoV is a significant infectious agent of captive felids because over half of the animals tested were positive by viral genetic detection, serology, or both. Dependence upon one method for detection of infection is unreliable.


title: [Asymptomatic SARS-CoV-2 infection in children: a clinical analysis of 20 cases].

text: OBJECTIVE To study the clinical and epidemiological features of children with asymptomatic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. METHODS The clinical data of 20 children who were diagnosed with asymptomatic SARS-CoV-2 infection from January 20 to March 4, 2020 were analyzed. RESULTS Among the 20 children, there were 7 boys (35%) and 13 girls (65%), aged 8 months to 14 years (mean 8±5 years). All these children had no clinical manifestations and attended the hospital for an epidemiological history of SARS-CoV-2. Nineteen children were shown with family aggregation of SARS-CoV-2 infection. Nasopharyngeal swabs were PCR-positive for SARS-CoV-2 in all 20 children. There were 4 children (20%) of mild type, 16 children (80%) of common type, and no children of severe type or critical type. The mean peripheral blood leukocyte count was (6.8±3.5)×109/L, and 7 children had an abnormal peripheral blood leukocyte count, with an increase in 5 children and a reduction in 2 children. One child had a decreased absolute value of lymphocytes (0.87×109/L), 3 children had an increased erythrocyte sedimentation rate (20-42 mm/h), 7 children had an increased lactate dehydrogenase level (>400 U/L), and 4 children had an increased blood lactate level (>1.6 mmol/L). Chest CT showed single or multiple small nodule shadows, patchy shadows, and ground-glass shadows in the middle or lateral lobe of lungs or under the pleura in 13 children. CONCLUSIONS Pediatric cases of asymptomatic SARS-CoV-2 infection mostly occur with family aggregation. Most of the children with asymptomatic infection have no obvious abnormalities in blood routine and other laboratory tests. Changes in chest CT scan can be used as an aid for early diagnosis of asymptomatic infection in children.


title: Thyroid activity in patients with major depression.

text: Hypothalamus-pituitary-thyroid (HPT) axis dysfunction has been associated with pathophysiology of major depression. The aim of the study was to determine serum levels of total 3,5,3'-triiodothyronine (T3), total thyroxine (T4) and thyroid-stimulating-hormone (TSH) in patients with major depression and healthy controls. The study included 53 medication-free patients with depression and 49 healthy controls. Exclusion criteria for patients was: other axis-I and axis-II diagnoses, intensive psychotherapy or electroconvulsive therapy, prior clinical and/or laboratory evidence of hypo- or hyperthyroidism, alcohol or nicotine dependence, pregnancy, hormone supplement therapy, somatic illnesses (diabetes, renal or hepatic disorders), infections or autoimmune diseases, recent surgical treatment or significantly changed body weight. For controls: the presence of psychiatric disorders and/or thyroid dysfunctions. The diagnosis of major depression was made using structured clinical interview based on DSM-IV criteria. The results showed significantly lower T3 and TSH levels in patients compared to controls. There was no significant difference in T4 values between patients with depression and control subjects. The results showing altered levels of thyroid hormones in depression indicate that further research on thyroid hormone activity can contribute to the better understanding of the biological basis of depression. Based on the high frequency of the subtle neuroendocrine disorders coexisting with depression, the association of thyroid abnormalities and depression should not be underestimated. Future research should identify different behavioral endophenotypes characteristic for depression, which would greatly facilitate delineating the biological phenomena associated with this psychiatric illness.


title: [Management of gastrointestinal and hepatic diseases during the COVID-19 outbreak].

text: The current epidemic of SARS-CoV-2 infection poses new challenges in the management of patients with gastrointestinal or liver disease. Consultations with patients with chronic diseases should ideally be done via telemedicine and treatments administered at home if possible. The latter should be maintained in non-infected subjects to limit the risk of decompensation of their underlying disease. In the event of proven infection, immunomodulatory or biological treatments will tend to be reduced or discontinued unless the disease is in a severely active phase. Elective endoscopy should be postponed, and urgent procedures should be performed with appropriate personal protective equipment.


title: Local and systemic cell-mediated immunity against transmissible gastroenteritis, an intestinal viral infection of swine.

text: The occurrence of local and systemic cell-mediated immunity (CMI) in swine against transmissible gastroenteritis (TGE) virus, a coronavirus that infects the epithelial cells of the small intestine of swine, has been investigated. Leukocyte suspensions prepared from intestinal lamina propria (LP) and spleen were each incubated with and without inactivated TGE virus. The supernatants from these cultures were tested for the presence of macrophage migration inhibition factor (MIF) by the indirect MIF test using guinea pig peritoneal exudate cells as the source of macrophages. All swine serologically negative for TGE antibodies (i.e., unexposed to TGE virus) were also negative for MIF production by lymphocytes from both sources. In 10 of 10 animals given TGE virus subcutaneously, MIF production by splenic lymphocytes was significantly greater (p less than or equal to 0.01) than by LP lymphocytes, and in 9 of 10 of these animals MIF production by LP lymphocytes was negligible (percentage of inhibition of migration less than 3%).


title: The immunological synapse for B-cell memory: the role of the ICOS and its ligand for the longevity of humoral immunity.

text: RECENT FINDINGS The ICOS is one of the many costimulatory molecules of T cells. Recently, the authors discovered that the homozygous loss of the ICOS in humans presents as common variable immunodeficiency, the most prevalent treated primary immunodeficiency in man. Therefore, the ICOS is after CD40 and CD40-ligand deficiency the third costimulatory molecule for which a human disease has been described. PURPOSE OF REVIEW The description of ICOS deficiency made it necessary to review the vast amount of knowledge from experimental models in the light of a new clinical immunodeficiency phenotype. To this end, after introducing the ICOS and its ligand and comparing the murine and human 'knockout', the role of the ICOS in parasitic and viral infections, asthma, autoimmunity, transplantation immunology, and tumor surveillance will be discussed. SUMMARY The focus of this review is the interaction between the ICOS and its ligand, which is essential for T cell dependent B cell responses.


title: A leader sequence is present on mRNA A of avian infectious bronchitis virus.

text: The subgenomic mRNAs of the fowl coronavirus infectious bronchitis virus (IBV) form a 3' co-terminal or 'nested' set. The RNase T1 oligonucleotide mapping data which demonstrated this form of sequence organization provided no evidence for the presence of non-contiguous (leader) sequences fused to the 5' termini of the mRNAs. However, we have been able to demonstrate, by extension of an end-labelled synthetic oligonucleotide primer, the presence of a leader sequence on IBV mRNA A. Sequencing by the chemical degradation method of the extended product has yielded the almost complete sequence of the 60 base leader and its point of fusion to the sequence of the virus genomic RNA.


title: Influenza surveillance in Indonesia: 1999-2003.

text: Although influenza is recognized for its worldwide importance, little is known about the disease from tropical countries like Indonesia. From August 1999 through January 2003, a surveillance study was conducted in clinics at 6 sentinel locations. Adults (age, >14 years) and children (age, 4-14 years) presenting with respiratory symptoms suggestive of influenza were asked to enroll in the study. Nasal and pharyngeal swabs were examined by virus isolation, polymerase chain reaction, and rapid immunochromatographic tests. A total of 3079 specimens were collected from 1544 participants. Influenza infection was confirmed in 172 volunteers (11.1%) presenting with influenza-like illness. Influenza A (H1N1 and H3N2) and B viruses were detected at all sites. Peak prevalence tended to coincide with the respective rainy seasons, regardless of location. In light of the recent epidemic of severe acute respiratory syndrome, continued influenza surveillance would be useful in strengthening the infrastructure of the Indonesian public health system.


title: High-throughput, sensitive, and accurate multiplex PCR-microsphere flow cytometry system for large-scale comprehensive detection of respiratory viruses.

text: Human respiratory viruses are a diverse group of pathogens composed of hundreds of virus strains, and this presents a major challenge for diagnostic laboratories. To efficiently detect numerous viruses in a large epidemiologic study, we developed a fast, multitarget, sensitive, and specific assay named the Respiratory MultiCode-PLx Assay (RMA). The RMA utilizes improved multiplex PCR chemistry (EraGen MultiCode-PLx technology) coupled with high-throughput microsphere flow cytometry (Luminex). Eighteen sets of virus-specific multiplex PCR primers were developed based on the conserved sequences of all available respiratory-virus sequences for eight distinct groups: human rhinovirus (HRV), respiratory syncytial virus (RSV), parainfluenza virus (PIV), influenza virus (InfV), metapneumovirus, adenovirus (Ad), coronavirus, and enterovirus. Each primer set detected 20 cDNA copies of the intended target per sample and had no reaction with 60,000 copies of human genomic DNA. The accuracy and sensitivity of the RMA for detecting respiratory viruses in human samples were tested with two sets of clinical specimens. First, 101 nasal-wash specimens that were positive for HRV, RSV, InfV, PIV, or Ad by traditional techniques were reanalyzed by RMA, and all target viruses were detected with an overall sensitivity of 94% and specificity of 99%. Second, 103 nasal-wash samples from 5-year-old children with asthma and respiratory symptoms were analyzed; RMA detected viruses in 74 specimens (71.8%) compared to only 24 (23.3%) by traditional culture and immunofluorescent-staining techniques. These results show that RMA is an accurate, sensitive, and practical test for respiratory-virus infections.


title: Coronavirus minus-strand RNA synthesis and effect of cycloheximide on coronavirus RNA synthesis.

text: The temporal sequence of coronavirus plus-strand and minus-strand RNA synthesis was determined in 17CL1 cells infected with the A59 strain of mouse hepatitis virus (MHV). MHV-induced fusion was prevented by keeping the pH of the medium below pH 6.8. This had no effect on the MHV replication cycle, but gave 5- to 10-fold-greater titers of infectious virus and delayed the detachment of cells from the monolayer which permitted viral RNA synthesis to be studied conveniently until at least 10 h postinfection. Seven species of poly(A)-containing viral RNAs were synthesized at early and late times after infection, in nonequal but constant ratios. MHV minus-strand RNA synthesis was first detected at about 3 h after infection and was found exclusively in the viral replicative intermediates and was not detected in 60S single-stranded form in infected cells. Early in the replication cycle, from 45 to 65% of the [3H]uridine pulse-labeled RF core of purified MHV replicative intermediates was in minus-strand RNA. The rate of minus-strand synthesis peaked at 5 to 6 h postinfection and then declined to about 20% of the maximum rate. The addition of cycloheximide before 3 h postinfection prevented viral RNA synthesis, whereas the addition of cycloheximide after viral RNA synthesis had begun resulted in the inhibition of viral RNA synthesis. The synthesis of both genome and subgenomic mRNAs and of viral minus strands required continued protein synthesis, and minus-strand RNA synthesis was three- to fourfold more sensitive to inhibition by cycloheximide than was plus-strand synthesis.


title: [Analysis of severe acute respiratory syndrome in Beijing].

text: OBJECTIVE To study the clinical, laboratory, and radiologic features of 34 cases of severe acute respiratory syndrome (SARS) in Beijing. METHODS All patients were admitted to the isolation wards. Their demographic, clinical, laboratory, and radiologic characteristics were analyzed. Univariate and multivariate analyses were performed. RESULTS Eight patients came from a family, and 15 patients were medical staff. The mean age of patients was (33.4 +/- 13.4) years. The latent period varied from 2 to 14 days (median 4 days). The most common symptoms were fever (100%), palpitation (91.7%), myalgia (79.2%), headache (70.8%), diarrhea (73.9%) and cough (58.3%). The mean leucocyte count was (4.6 +/- 1.4) x 10(9)/L, and the mean lymphocyte ratio was 0.27 +/- 0.11. 68.4% of the patients had lymphopenia (absolute lymphocyte count < 1.3 x 10(9)/L). Other common findings included elevated levels of serum alanine aminotransferase, lactate dehydrogenase and erythrocyte sedimentation (76.2%, 28.6% and 47.8%, respectively), and decreased levels of serum iron and albumin (63.2% and 47.8%, respectively). Thirty-two cases had abnormal chest radiographs. In 2 cases in whom typical lung opacities could not be found on the initial plain chest radiographs, thoracic CT proved to be useful. Postmortem examination of 1 patient revealed marked edema with foci of hemorrhage and hyaline membrane formation in the lungs, hemorrhage necrosis and a obvious decline of cells in lymph glands. In a multivariate analysis (Stata 7.0), the independent predictor of an adverse outcome was advanced age (odds ratio per decade of life, 1.6; 95% CI, 1.08 to 2.63; P = 0.007). CONCLUSIONS Fever, lymphopenia, low serum iron and chest radiograph are helpful to diagnose SARS early; age is the independent predictor of an outcome.


title: [Porcine aminopeptidase N is a functional receptor for the PEDV coronavirus].

text: Porcine epidemic diarrhea virus (PEDV) causes lethal diarrhea in piglets that leads to great economic losses in East Asia. It was reported that aminopeptidase N (APN) was the receptor for Transmissible gastroenteritis virus (TGEV), Human coronavirus 229E (HCoV-229E) and Feline coronavirus (FeCoV) which all belonged to group I coronavirus including PEDV. It was also confirmed previously that porcine aminopeptidase N (pAPN) could bind to PEDV, and anti-pAPN antibodies could inhibit the combination. To investigate whether pAPN was a receptor for PEDV, we transfected MDCK cells with porcine aminopeptidase (pAPN) cDNA and this enabled non-susceptible cells to support PEDV replication and serial viral propagation. Moreover, the infection was blocked by antibodies against pAPN, implying the critical role of pAPN during virus entry. In addition, immunofluorescence assays for detection of pAPN and PEDV antigens, together with neutralization assays using antibodies against pAPN, further confirmed the correlation between pAPN expression and viral replication in pAPN-transfected MDCK cells. These results indicated that pAPN is a functional receptor for PEDV.


title: Synthesis and processing of the bovine enteric coronavirus haemagglutinin protein.

text: The haemagglutinin molecule on the bovine enteric coronavirus has been identified as a glycoprotein of 140K composed of disulphide-linked subunits of 65K. In this study, we have shown the subunits to be identical by demonstrating an unambiguous amino-terminal amino acid sequence. The unglycosylated subunit was found to have an Mr of 42.5K and to undergo rapid disulphide linkage and glycosylation. Glycosylation was found to be of the asparagine-linked type and some of the oligosaccharides underwent processing to complex forms. Studies with inhibitors of glycosylation suggested that a processing of the haemagglutinin oligosaccharide takes place on the virion whilst it is in the Golgi apparatus. Each haemagglutinin subunit on the mature virion was estimated to possess six or seven carbohydrate chains of either the high-mannose or hybrid type, and three or four chains of the complex type.


Physical interaction between the membrane (M) and envelope (E) proteins of the coronavirus avian infectious bronchitis virus (IBV).


title: Naturally occurring anti-Escherichia coli protein antibodies in the sera of healthy humans cause analytical interference in a recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay for serodiagnosis of severe acute respiratory syndrome.

text: We reported the analytical interference of anti-Escherichia coli protein (EP) antibodies in human sera and residual EP in a recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay as a possible source of false positives in severe acute respiratory syndrome serodiagnosis. The rate of false positives was significantly reduced by adding mouse anti-EP antiserum in the blocking step.


title: The viral nucleocapsid protein of transmissible gastroenteritis coronavirus (TGEV) is cleaved by caspase-6 and -7 during TGEV-induced apoptosis.

text: The transmissible gastroenteritis coronavirus (TGEV), like many other viruses, exerts much of its cytopathic effect through the induction of apoptosis of its host cell. Apoptosis is coordinated by a family of cysteine proteases, called caspases, that are activated during apoptosis and participate in dismantling the cell by cleaving key structural and regulatory proteins. We have explored the caspase activation events that are initiated upon infection of the human rectal tumor cell line HRT18 with TGEV. We show that TGEV infection results in the activation of caspase-3, -6, -7, -8, and -9 and cleavage of the caspase substrates eIF4GI, gelsolin, and alpha-fodrin. Surprisingly, the TGEV nucleoprotein (N) underwent proteolysis in parallel with the activation of caspases within the host cell. Cleavage of the N protein was inhibited by cell-permeative caspase inhibitors, suggesting that this viral structural protein is a target for host cell caspases. We show that the TGEV nucleoprotein is a substrate for both caspase-6 and -7, and using site-directed mutagenesis, we have mapped the cleavage site to VVPD(359) downward arrow. These data demonstrate that viral proteins can be targeted for destruction by the host cell death machinery.


title: The pathogenesis of MHV nucleocapsid gene chimeric viruses.

text: A set of viruses in which various segments of the nucleocapsid (N) gene of MHV have been substituted with the corresponding segments of bovine coronavirus (BCV) by targeted recombination were analyzed for their biologic properties. Histology for organ pathology and plaque assay for viral titer analysis following intracerebral (IC) inoculation were studied. One chimeric virus (Alb85), in which only a small segment of the N gene was replaced, exhibited a phenotype similar to wild type MHV-A59. However, three of the chimeric viruses (Alb106, Alb112 and Alb100) produced acute encephalitis and demyelination but without hepatitis following IC inoculation. Intravenous (IV) and intrahepatic (IH) inoculations were able to restore the ability of these viruses to produce hepatitis. The common denominator of the three chimeric viruses with a different phenotype is a region between aa 306 and aa 386 in which 17 amino acids (aa) differences exist between the two strains. Thus this region may contain determinants which enable the virus to exit the brain and reach the blood stream.


title: Expression and functional characterisation of the putative SARS coronavirus non-structural proteins X1-X5.

text: 1. We produced mammalian expression vectors encoding the SARS coronavirus (SARS-CoV) accessory proteins with or without the fluorescence protein tag and cell lines with stable expression of these proteins. 2. The cellular localisation and function of the SARS-CoV accessory proteins was determined. 3. SARS 6 and SARS 8b proteins are localised to the endoplasmic reticulum and nucleus/cytoplasm, respectively, and both proteins stimulate host cell DNA synthesis.


title: The psychological effect of severe acute respiratory syndrome on emergency department staff.

text: BACKGROUND The severe acute respiratory syndrome (SARS) outbreak in 2003 affected 29 countries. The SARS outbreak was unique in its rapid transmission and it resulted in heavy stress in first-line healthcare workers, particularly in the emergency department. AIM : To determine the influence of SARS on the psychological status, including post-traumatic stress disorder (PTSD) symptoms, of the staff in the emergency department. METHODS To investigate whether different working conditions in the hospital led to different psychological effects on healthcare workers, the psychological effect on emergency department staff in the high-risk ward was compared with that on psychiatric ward staff in the medium-risk ward. Davidson Trauma Scale-Chinese version (DTS-C) and Chinese Health Questionnaire-12 (CHQ-12) items were designed to check the psychological status of the staff in the month after the end of the SARS outbreak. RESULTS 86 of 92 (93.5%) medical staff considered the SARS outbreak to be a traumatic experience. The DTS-C scores of staff in the emergency department and in the psychiatric ward were significantly different (p = 0.04). No significant difference in CHQ score was observed between the two groups. Emergency department staff had more severe PTSD symptoms than staff in the psychiatric ward. CONCLUSION SARS was a traumatic experience for healthcare providers in Taiwan. Most staff in the emergency department and in the psychiatric ward had PTSD. Emergency department staff had more severe PTSD symptoms than staff in the psychiatric ward.


docid: uvpdbssy

title: Intra-articular versus serum C-reactive protein analysis in suspected periprosthetic knee joint infection.

text: The purpose of this study was to investigate the comparative diagnostic strength of C-reactive protein (CRP) values in synovial fluid and serum in patients with presumed periprosthetic joint infection. We collected 44 synovial fluid samples and 24 serum samples from 43 patients. Patients were judged to be uninfected or infected based on standardized criteria. Synovial and serum samples were obtained simultaneously. We determined the diagnostic strength of our assay by using receiver operating characteristic curve analysis, calculated the cutoff value and calculated the difference in diagnostic strength for periprosthetic joint infection between synovial fluid and blood serum analysis. The area under the curve was 0.977 for intra-articular CRP analysis and 0.979 for serum CRP analysis. The cutoff points for intra-articular CRP analysis where 1.8 mg/L and 2.8 mg/L. Both tests showed a very high diagnostic strength; the difference in diagnostic strength between synovial fluid and blood serum analysis was not significant (p = 0.66).


title: Characteristics of porcine circovirus-2 replication in lymphoid organs of pigs inoculated in late gestation or postnatally and possible relation to clinical and pathological outcome of infection.

text: In this study, the characteristics of porcine circovirus-2 (PCV2) replication (infectious virus titrations, distribution, and immunophenotyping of infected cells) in lymphoid organs were examined and related to the development of clinical signs and histological lesions in 26 piglets that had been inoculated with PCV2 either in utero or at 1 day of age. Piglets inoculated in utero at 92 or 104 gestational days (n = 12) were collected by Caesarean section at term and either sacrificed immediately or kept in isolators and allowed to live postnatally until 35 days postinoculation (PI). Caesarean-derived piglets inoculated at 1 day of age (n = 14) were sacrificed at 10, 21, 35, 42, and 49 days PI. Spleen and lymph nodes were collected for virologic and histopathological examinations. Clinical signs were not observed in any of the piglets. High virus titers (10(4.5-5.7) TCID50/g [TCID refers to tissue culture infectious dose]) were detected in 6 of the 26 piglets. Three of these 6 piglets were euthanized at 10 days PI, and infected cells of the monocyte-macrophage lineage (SWC3+, CD14+, and sialoadhesin [Sa]+ cells) and infected cells bearing lymphocyte markers (CD4+, CD8+, and immunoglobulin M+ cells) were identified by double-immunofluorescence labeling on serial cryostat sections. The other 3 piglets were euthanized at 21 and 35 days PI, and the majority of infected cells were SWC3+, CD14+, and Sa-. The absence of Sa in these infected cells, together with their localization in lymphocyte-dependent regions, suggests that they were infiltrating monocytic cells. Sialoadhesin is highly expressed in differentiated macrophages and not in peripheral blood mononuclear cells. In all 6 piglets with high virus titers, lymphocyte depletion and infiltration of monocytic cells were observed. In the remaining 20 piglets with virus titers less than 10(4.5) TCID50/g, the majority of infected cells were SWC3+, CD14+, and Sa+. In conclusion, it can be stated that high PCV2 titers in lymphoid organs may lead to the development of histological lesions similar to those observed in pigs with postweaning multisystemic wasting syndrome without causing disease. Furthermore, in lymphoid organs with high virus titers, infection occurs mainly in infiltrating monocytic cells and to a limited extent in cells bearing lymphocyte markers.


title: Bacterial carriage and respiratory tract infections in subjects > or = 60 years during an influenza season: implications for the epidemiology of community acquired pneumonia and influenza vaccine effectiveness.

text: INTRODUCTION During the 2010/11 influenza season an epidemiological prospective cohort active study was organized, to evaluate etiologic role due to the main bacteria and viruses causing Community Acquired Pneumonia (CAP) and Influenza like-illness (ILl) in elderly and to explore the role of the bacterial nose-pharingeal carriage in subjects with respiratory tract infections. METHODS An integrated active surveillance of a cohort of adults aged > or = 60 y based on a double prospective and retrospective mechanisms of capture of ILl and CAP cases was organized. Samples were collected from all ILI and CAP prospectively identified. The samples were be tested by multiplex PCR for detection of the main respiratory bacteria and viruses. RESULTS AND DISCUSSION The study population amounted to 2,551 adults. During the 2010/11 influenza season, the ILl cumulative incidence was 4.2%, that was twice higher than that calculated by regional sentinel-based Influenza surveillance system during the 2010/11 season in the elderly (2.2%). Among 45 patients with ILI of which had been collected the swab, 17 (37.8%) were positive for influenza viruses and 2 (4.4%) for RSV, 6 (13.3%) patients carried Streptococcus pn and 6 (13.3%) Haemophilus in. In the same period, 7 CAP cases were observed; 3 cases were prospectively identified and samples were collected, while 4 cases were retrospectively detected. The CAP cumulative incidence was 0,3%. The influenza vaccine effectiveness in prevention of laboratory-confirmed influenza emerged by our study was 61%, in condition of good antigenic matching between vaccine and circulating strains observed during the 2010/11. These data contribute to better defining the epidemiological picture of upper and lower respiratory tract infections, fundamental information in light of the recent introduction of new vaccines for prevention of pneumonia in the elderly, including 13-valent conjugate pneumococcal vaccine.


title: Integrating nutrition with ecology: balancing the health of humans and biosphere.

text: OBJECTIVE To show that current rates of global population growth, production and consumption of food, and use of living and physical resources, are evidently not sustainable. To consider ways in which nutrition and allied sciences can respond to this great challenge of the twenty-first century. METHOD Past, current and future projected trends in production and consumption patterns are examined. These show that overall present and projected patterns cannot be sustained; and also show increasing unacceptable inequity between and within rich and poor regions and countries. DISCUSSION Nutrition science classically focuses on nutrients in relation to human physiology, metabolism, growth, health and disease. The social and environmental conditions of the modern, interconnected, market-oriented world, and the consequences for food production and consumption, are extending the research and policy agenda with which nutrition science must now urgently engage. Historically, much attention has been paid to eliminating nutritional deficiency states, and this remains an important task. In modern urban populations 'malnutrition' encompasses new forms of dietary imbalance, especially excesses of certain nutrients. These contribute to various non-communicable diseases and, particularly, to overweight/obesity and its attendant metabolic derangements and disease risks. As a mass phenomenon the current surge in obesity has no historical precedent. The escalating impact of humankind on the natural environment, with its ramifications for present and future food production, is also unprecedented. CONCLUSION The essential challenge for nutrition science is to develop new understanding and strategies to enable a balance between promoting, equitably, the health of humans while sustaining the long-term health of the biosphere. Extension of nutrition science and food policy to meet those goals will be aided by understanding better how dietary conditions shaped the biological evolution of humankind. The fundamental long-term task is to integrate human health with the health of the biosphere.


title: [Effect on mRNA and secretion levels of proinflammatory cytokines in DC infected by SARS-CoV N gene recombinant adenovirus].

text: OBJECTIVE To investigate the exact mechanism of SARS-CoV pathogenesis at the protein level. METHOD Under the condition of the establishment of dendrtic cells (DC) culture method, we used recombinant adenovirus to infect mature DC to make clear the development changes in mRNA levels and secreted protein levels of proinflammatory cytokines of IL-1beta, IL-6, IL-8 and TNF-alpha by using RT-PCR and ELISA. RESULT We found that mRNA levels and secreted protein levels of IL-6 and TNF-alpha in DC had increased gradually after rAd-N infection during first 24 h compared with the control DC infected by rAd-LacZ (P < 0.05 or P < 0.01). CONCLUSION It is suggested that N protein may be related to the excessive secretion of proinflammatory cytokines during SARS-CoV infection at the acute phase.


title: Recognition of mRNA cap structures by viral and cellular proteins.

text: Most cellular and eukaryotic viral mRNAs have a cap structure at their 5' end that is critical for efficient translation. Cap structures also aid in mRNA transport from nucleus to cytoplasm and, in addition, protect the mRNAs from degradation by 5' exonucleases. Cap function is mediated by cap-binding proteins that play a key role in translational control. Recent structural studies on the cellular cap-binding complex, the eukaryotic translation initiation factor 4E and the vaccinia virus protein 39, suggest that these three evolutionary unrelated cap-binding proteins have evolved a common cap-binding pocket by convergent evolution. In this pocket the positively charged N(7)-methylated guanine ring of the cap structure is stacked between two aromatic amino acids. In this review, the similarities and differences in cap binding by these three different cap-binding proteins are discussed. A comparison with new functional data for another viral cap-binding protein--the polymerase basic protein (PB2) of influenza virus--suggests that a similar cap-binding mechanism has also evolved in influenza virus.


title: Monkeypox in the United States: an occupational health look at the first cases.

text: Between May 15 and June 20, 2003, 71 suspected cases of monkeypox were investigated and 37 individuals in the United States developed laboratory confirmed monkeypox. These were the first cases of human monkeypox ever documented in the United States or in the Western Hemisphere. The disease was transmitted from small animals imported from Africa to other animals, including prairie dogs sold as pets throughout the U.S. Midwest. Direct contact with the infected animals was the method of infection, and although human to human transmission was thought to have occurred, this was not confirmed by follow up testing. Because of the link with contact with a prairie dog, initial evaluation of the disease was focused toward diseases commonly associated with this animal (e.g., tularemia, plague). Laboratory findings at the Marshfield Clinic in Marshfield, Wisconsin pointed to the presence of an orthopox. The CDC confirmed monkeypox was the infecting orthopox agent. Occupational health nurses from the Marshfield Clinic had direct involvement in the identification and follow up of employees who had direct contact with the diagnosed patients. Programs, such as a respiratory protection program initiated and carried out by Clinic occupational health nurses, were used to prevent employee exposure for Clinic staff. One Clinic employee was thought to potentially have monkeypox because of her direct contact with one of the patients. Four Clinic employees were vaccinated with vaccinia vaccine as a result of their contact with patients or lab specimens. Quarantine of the potentially infected employee and her boyfriend uncovered issues that must be addressed if other infectious diseases requiring quarantine or isolation of individuals emerge or re-emerge. These include a system to compensate individuals in quarantine or isolation who do not have any other source of income. The issue of whether workers' compensation should cover an employee who is quarantined or isolated for a potential work related exposure to an infectious disease if no disease is actually diagnosed also needs to be explored. A better system of getting state or CDC laboratory results back to the local level, including the occupational health area of the generating facility, must be developed. This will be very important if diseases such as severe acute respiratory syndrome (SARS) or smallpox should re-emerge in the United States. Occupational health nurses are an integral part of any infectious disease process occurring in the United States. The identification of monkeypox in the United States shows that any planning to detect, prevent, and treat diseases with the potential to affect the employee population must include occupational health nurse involvement.


title: Cell-penetrating peptide-morpholino conjugates alter pre-mRNA splicing of DMD (Duchenne muscular dystrophy) and inhibit murine coronavirus replication in vivo.

text: The cellular uptake of PMOs (phosphorodiamidate morpholino oligomers) can be enhanced by their conjugation to arginine-rich CPPs (cell-penetrating peptides). Here, we discuss our recent findings regarding (R-Ahx-R)(4)AhxB (Ahx is 6-aminohexanoic acid and B is beta-alanine) CPP-PMO conjugates in DMD (Duchenne muscular dystrophy) and murine coronavirus research. An (R-Ahx-R)(4)AhxB-PMO conjugate was the most effective compound in inducing the correction of mutant dystrophin transcripts in myoblasts derived from a canine model of DMD. Similarly, normal levels of dystrophin expression were restored in the diaphragms of mdx mice, with treatment starting at the neonatal stage, and protein was still detecTable 22 weeks after the last dose of an (R-Ahx-R)(4)AhxB-PMO conjugate. Effects of length, linkage and carbohydrate modification of this CPP on the delivery of a PMO were investigated in a coronavirus mouse model. An (R-Ahx-R)(4)AhxB-PMO conjugate effectively inhibited viral replication, in comparison with other peptides conjugated to the same PMO. Shortening the CPP length, modifying it with a mannosylated serine moiety or replacing it with the R(9)F(2) CPP significantly decreased the efficacy of the resulting PPMO (CPP-PMO conjugate). We attribute the success of this CPP to its stability in serum and its capacity to transport PMO to RNA targets in a manner superior to that of poly-arginine CPPs.


Severe acute respiratory syndrome and biology, air quality, physics, and mechanical engineering.


title: Feasibility of a Modified Biopsy Needle for Irreversible Electroporation Ablation and Periprocedural Tissue Sampling.

text: OBJECTIVES To test the feasibility of modified biopsy needles as probes for irreversible electroporation ablation and periprocedural biopsy. METHODS Core biopsy needles of 16-G/9-cm were customized to serve as experimental ablation probes. Computed tomography-guided percutaneous irreversible electroporation was performed in in vivo porcine kidneys with pairs of experimental (n = 10) or standard probes (n = 10) using a single parameter set (1667 V/cm, ninety 100 µs pulses). Two biopsy samples were taken immediately following ablation using the experimental probes (n = 20). Ablation outcomes were compared using computed tomography, simulation, and histology. Biopsy and necropsy histology were compared. RESULTS Simulation-suggested ablations with experimental probes were smaller than that with standard electrodes (455.23 vs 543.16 mm2), although both exhibited similar shape. Computed tomography (standard: 556 ± 61 mm2, experimental: 515 ± 67 mm2; P = .25) and histology (standard: 313 ± 77 mm2, experimental: 275 ± 75 mm2; P = .29) indicated ablations with experimental probes were not significantly different from the standard. Histopathology indicated similar morphological changes in both groups. Biopsies from the ablation zone yielded at least 1 core with sufficient tissue for analysis (11 of the 20). CONCLUSIONS A combined probe for irreversible electroporation ablation and periprocedural tissue sampling from the ablation zone is feasible. Ablation outcomes are comparable to those of standard electrodes.


title: Recombinant truncated nucleocapsid protein as antigen in a novel immunoglobulin M capture enzyme-linked immunosorbent assay for diagnosis of severe acute respiratory syndrome coronavirus infection.

text: We report the development of an immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) for severe acute respiratory syndrome coronavirus (SARS-CoV) by using recombinant truncated SARS-CoV nucleocapsid protein as the antigen. The newly developed MAC-ELISA had a specificity and sensitivity of 100% as evaluated by using sera from healthy volunteers and patients with laboratory-confirmed SARS. Using serial serum samples collected from SARS patients, the times to seroconversion were determined by IgM antibody detection after SARS-CoV infection. The median time to seroconversion detection was 8 days (range, 5 to 17 days) after disease onset, and the seroconversion rates after the onset of illness were 33% by the first week, 97% by the second week, and 100% by the third week. Compared with the results of our previous report on the detection of IgG, the median seroconversion time by IgM detection was 3 days earlier and the seroconversion rate by the second week after the illness for IgM was significantly higher than by IgG assay. Our results indicating that the IgM response appears earlier than IgG after SARS-CoV infection in consistent with those for other pathogens. Our newly developed MAC-ELISA system offers a new alternative for the confirmation of SARS-CoV infection.


title: [Isolation and identification of SARS-coronavirus in nasal and throat swabs collected from clinically diagnosed SARS patients].

text: OBJECTIVE To isolate and identify SARS-coronavirus in nasal and throat swabs collected from clinically diagnosed severe acute respiratory syndrome (SARS) patients. METHODS Nasal and throat swab specimens were inoculated onto well of 24-well plate containing confluent monolayers of Vero and MRC-5 cells. Isolates were identified with serology, electron microscopy and genome sequence. RESULTS One hundred and fifty-eight nasal and throat swabs specimens from 79 SARS patients in Peking Union Medical College Hospital between April and May, 2003 were cultured for SARS-coronavirus. Cytopathic effect (CPE) was found in three nasal swab specimens inoculated in Vero cells. Acute and convalescent phase serum specimens collected from SARS patients were found with seroconversions and/or a fourfold or greater rises in indirect fluorescence antibodies (IgG and IgM) titers when the 3 isolates (infected Vero cells) were used as antigen. Coronavirus was observed in the culture supernatant by negative-stain electron microscopy. Genome sequence confirmed the isolates were SARS-coronavirus. CONCLUSIONS The 3 isolates from nasal and throat swabs samples collected from 79 clinically diagnosed SARS patients were SARS coronavirus.


title: Simultaneous detection of canine respiratory disease associated viruses by a multiplex reverse transcription-polymerase chain reaction assay.

text: A multiplex reverse transcription polymerase chain reaction (mRT-PCR) assay was developed for the simultaneous detection of canine distemper virus (CDV), canine respiratory coronavirus (CRCoV) and canine influenza virus (CIV). These viral pathogens are all causative agents of canine infectious respiratory disease (CIRD). The sensitivity and specificity of the mRT-PCR were determined by comparing it to a rapid antigen test (RAT) or immuno-chromatography test kit and reverse transcription-polymerase chain reaction (RT-PCR) in the detection of CDV, CRCoV and CIV antigens present in 100 clinical samples (nasal swabs and whole blood samples) from 50 dogs with respiratory disease symptoms. This study revealed that mRT-PCR had almost exactly the same performance or results were almost 100% in agreement with that of RT-PCR and RAT both in terms of the assay sensitivity and specificity which was more highly evident in detecting CIV, CDV and CRCoV antigens present in canine nasal swab samples. Therefore, this assay could be a better alternative for the definitive and simultaneous ante-mortem detection of the three viral pathogens that cause CIRD by using nasal swabs.


title: Virus-specific antibody, in the absence of T cells, mediates demyelination in mice infected with a neurotropic coronavirus.

text: Mice infected with mouse hepatitis virus strain JHM develop an inflammatory demyelinating disease in the central nervous system with many similarities to human multiple sclerosis. The mouse disease is primarily immune-mediated because demyelination is not detected in JHM-infected mice lacking T or B cells but does occur after transfer of JHM-specific T cells. Although less is known about the ability of antibodies to mediate demyelination, the presence of oligoclonally expanded B cells and high concentrations of antibodies (against self or infectious agents) in the central nervous system of many multiple sclerosis patients suggests that antibodies may also contribute to myelin destruction. Here, we show that anti-JHM antibodies, in the absence of T or B cells, caused demyelination in JHM-infected mice. Anti-JHM antibody was detected adjacent to areas of demyelination, consistent with a direct interaction between antibody and infected cells. Demyelination was reduced by 85 to 90% in infected RAG1(-/-) mice lacking normal expression of activating Fc receptors (FcRgamma(-/-)) and by approximately 76% when complement was depleted by treatment with cobra venom factor. These data demonstrate that JHM-specific antibodies are sufficient to cause demyelination and that myelin destruction in the presence of anti-virus antibodies results from a combination of complement- and Fc receptor-dependent mechanisms.


title: Conserved receptor-binding domains of Lake Victoria marburgvirus and Zaire ebolavirus bind a common receptor.

text: The GP(1,2) envelope glycoproteins (GP) of filoviruses (marburg- and ebolaviruses) mediate cell-surface attachment, membrane fusion, and entry into permissive cells. Here we show that a 151-amino acid fragment of the Lake Victoria marburgvirus GP1 subunit bound filovirus-permissive cell lines more efficiently than full-length GP1. An homologous 148-amino acid fragment of the Zaire ebolavirus GP1 subunit similarly bound the same cell lines more efficiently than a series of longer GP1 truncation variants. Neither the marburgvirus GP1 fragment nor that of ebolavirus bound a nonpermissive lymphocyte cell line. Both fragments specifically inhibited replication of infectious Zaire ebolavirus, as well as entry of retroviruses pseudotyped with either Lake Victoria marburgvirus or Zaire ebolavirus GP(1,2). These studies identify the receptor-binding domains of both viruses, indicate that these viruses utilize a common receptor, and suggest that a single small molecule or vaccine can be developed to inhibit infection of all filoviruses.


title: Middle East respiratory syndrome novel corona MERS-CoV infection. Epidemiology and outcome update.

text: Middle East respiratory syndrome coronavirus MERS-CoV is a newly emerging respiratory virus with a high case fatality rate among identified cases. The virus is thought to cause a severe disease in patients with underlying comorbidities. The identification of asymptomatic patients and mild cases among family and healthcare worker contacts of confirmed cases indicates a wider spectrum of clinical manifestation of the disease. The majority of patients presented with fever 98%, fever with cough 83%, and shortness of breath 72%. Radiographic manifestations range from unilateral infiltrate 43%, to increased bronchovascular markings 17%, and diffuse reticulonodular pattern 4%. Our understanding of the epidemiology and clinical presentation of the disease is increasing overtime. It is still not known what the source of the virus is and what the best treatment modality should be.


Cardiac Tamponade from Coronary Artery Injury after Laparoscopic Anti-Reflux Surgery.


title: Sabadinine: a potential non-peptide anti-severe acute-respiratory-syndrome agent identified using structure-aided design.

text: A novel human coronavirus has been reported to be the causative agent of severe acute respiratory syndrome (SARS). Since replication of HcoVs depends on extensive proteolytic processing, the main proteinase, 3CLpro, is an attractive drug target for anti-SARS agents. We have employed molecular docking of a chemical database into the active site of 3CLpro to search for non-peptidyl inhibitors. One compound was identified to be the natural product sabadinine, isolated from a historical herbal remedy.


title: Virus maturation by budding.

text: Enveloped viruses mature by budding at cellular membranes. It has been generally thought that this process is driven by interactions between the viral transmembrane proteins and the internal virion components (core, capsid, or nucleocapsid). This model was particularly applicable to alphaviruses, which require both spike proteins and a nucleocapsid for budding. However, genetic studies have clearly shown that the retrovirus core protein, i.e., the Gag protein, is able to form enveloped particles by itself. Also, budding of negative-strand RNA viruses (rhabdoviruses, orthomyxoviruses, and paramyxoviruses) seems to be accomplished mainly by internal components, most probably the matrix protein, since the spike proteins are not absolutely required for budding of these viruses either. In contrast, budding of coronavirus particles can occur in the absence of the nucleocapsid and appears to require two membrane proteins only. Biochemical and structural data suggest that the proteins, which play a key role in budding, drive this process by forming a three-dimensional (cage-like) protein lattice at the surface of or within the membrane. Similarly, recent electron microscopic studies revealed that the alphavirus spike proteins are also engaged in extensive lateral interactions, forming a dense protein shell at the outer surface of the viral envelope. On the basis of these data, we propose that the budding of enveloped viruses in general is governed by lateral interactions between peripheral or integral membrane proteins. This new concept also provides answers to the question of how viral and cellular membrane proteins are sorted during budding. In addition, it has implications for the mechanism by which the virion is uncoated during virus entry.


Regulation of MHC class I and II antigens on cerebral endothelial cells and astrocytes following MHV-4 infection.