title: Severe acute respiratory syndrome: providing care while minimizing personal risks.

text: Severe acute respiratory syndrome (SARS) poses a threat to most countries because of easy and convenient travel across the globe in a matter of hours. SARS is a recently recognized infectious disease that may lead to death and severe pulmonary sequelae. It presents with non-specific signs and symptoms and because no definitive laboratory test is readily available, it poses a great risk to healthcare workers because quarantine of affected individuals may not occur early in its presentation. Healthcare workers should consider the diagnosis and institute prompt and thorough protective measures in suspected cases.


title: The sialate-4-O-acetylesterases of coronaviruses related to mouse hepatitis virus: a proposal to reorganize group 2 Coronaviridae.

text: Group 2 coronaviruses are characterized within the order Nidovirales by a unique genome organization. A characteristic feature of group 2 coronaviruses is the presence of a gene encoding the haemagglutinin-esterase (HE) protein, which is absent in coronaviruses of groups 1 and 3. At least three coronavirus strains within group 2 expressed a structural protein with sialate-4-O-acetylesterase activity, distinguishing them from other members of group 2, which encode an enzyme specific for 5-N-acetyl-9-O-acetylneuraminic acid. The esterases of mouse hepatitis virus (MHV) strains S and JHM and puffinosis virus (PV) specifically hydrolysed 5-N-acetyl-4-O-acetylneuraminic acid (Neu4,5Ac2) as well as the synthetic substrates p-nitrophenyl acetate, 4-methylumbelliferyl acetate and fluorescein diacetate. The K(m) values of the MHV-like esterases for the latter substrates were two- to tenfold lower than those of the sialate-9-O-acetylesterases of influenza C viruses. Another unspecific esterase substrate, alpha-naphthyl acetate, was used for the in situ detection of the dimeric HE proteins in SDS-polyacrylamide gels. MHV-S, MHV-JHM and PV bound to horse serum glycoproteins containing Neu4,5Ac2. De-O-acetylation of the glycoproteins by alkaline treatment or incubation with the viral esterases resulted in a complete loss of recognition, indicating a specific interaction of MHV-like coronaviruses with Neu4,5Ac2. Combined with evidence for distinct phylogenetic lineages of group 2 coronaviruses, subdivision into subgroups 2a (MHV-like viruses) and 2b (bovine coronavirus-like viruses) is suggested.


title: The role of biosensors in the detection of emerging infectious diseases.

text: Global biosecurity threats such as the spread of emerging infectious diseases (i.e., avian influenza, SARS, Hendra, Nipah, etc.) and bioterrorism have generated significant interest in recent years. There is considerable effort directed towards understanding and negating the proliferation of infectious diseases. Biosensors are an attractive tool which have the potential to detect the outbreak of a virus and/or disease. Although there is a host of technologies available, either commercially or in the scientific literature, the development of biosensors for the detection of emerging infectious diseases (EIDs) is still in its infancy. There is no doubt that the glucose biosensor, the gene chip, the protein chip, etc. have all played and are still playing a significant role in monitoring various biomolecules. Can biosensors play an important role for the detection of emerging infectious diseases? What does the future hold and which biosensor technology platform is suitable for the real-time detection of infectious diseases? These and many other questions will be addressed in this review. The purpose of this review is to present an overview of biosensors particularly in relation to EIDs. It provides a synopsis of the various types of biosensor technologies that have been used to detect EIDs, and describes some of the technologies behind them in terms of transduction and bioreceptor principles.


title: Influence of viral and bacterial respiratory infections on exacerbations and symptom severity in childhood asthma.

text: The recent development of PCR for the diagnosis of respiratory viral infection has highlighted the importance of these infections in acute exacerbations of asthma. Rhinoviruses are important in all age groups, but particularly over 1 yr, while the role of RSV in bronchiolitis and wheezing in infants has been reaffirmed. Recent studies using the same technique for the detection of C. pneumoniae suggest a high prevalence of chronic infection in asthmatic children, and that the immune response to this organism may play a pathological role. These studies now require confirmation with larger carefully controlled studies.


title: Survey of Respiratory Virus in Patients Hospitalised for Acute Exacerbations of Heart Failure - A Prospective Observational Study.

text: INTRODUCTION Respiratory virus (RV) infections have been implicated in acute exacerbation cardiopulmunary conditions. This study aimed to determine the prevalence of RV infections in patients admitted to the cardiology unit with acute decompensated heart failure (ADHF) in a tertiary hospitals in Singapore. MATERIALS AND METHODS This was a single-centre, prospective observational study. A total of 194 adults (aged >21) admitted to the Singapore General Hospital with ADHF were recruited. A nasopharyngeal swab was taken for multiplex polymerase chain reaction (PCR) detection of influenza virus, rhinovirus, parainfluenza virus (HPIV), human coronavirus (HcoV), adenoviurs, human bocavirus (HboV), human metapneumovirus (hMPV), and respiratory syncytial virus (RSV). RESULTS Twenty-five (13%) had RVs detected by RV multiplex PCR. There comprised 9 rhinoviruses (36%), 4 influenza A viruses (16%), 3 HPIV (12%), 3 HCoV (12%), 2 adenoviruses (8%), 1 human HBoV (4%), 1 hMPV (4%), and 1 RSV (4%). Symptoms-wise, cough was significantly more common in the PCR-positive group (48% vs 24%, P = 0.02). There were no statistically significant differences in laboratory investigations (haemoglobin, leukocytes, platelets, creatine kinase, creatine kinase-muscle/brain, troponin T), and radiology findings between RV PCR-positive and -negative groups. The PCR-positive group did not have increased mortality or length of hospital stay. CONCLUSION This study identified a considerable burden of RVs in our ADHF cohort, and highlights the need for prevention of RVs in this group of patients. We also recognised the difficulty with clinical diagnosis of RVs in ADHF patients.


title: Public hospital preparations for SARS outbreak: experience of Alexandra Hospital.

text: Alexandra Hospital (AH) was one of the public hospitals in Singapore that responded to the severe acute respiratory syndrome (SARS) crisis. Being the only public hospital that remained "SARS-free", i.e., with no documented intra-institutional spread of disease, AH had to tackle a sudden, two-fold surge in hospital attendances and patient volume. Being the oldest hospital with a traditional open ward design and lack of proper isolation facilities, tough command and control policies had to be implemented to reduce the risk of a SARS outbreak. Stringent infection control measures, screening and triage, clinical procedures, and administrative policies all were important factors in helping the hospital balance the need to run routine operations while "fighting" SARS. Staff and people management also were crucial in keeping the workforce healthy and maintaining their morale and confidence during this difficult period.


docid: onahmjht

title: Cloning and sequencing of 5' terminal sequences from avian infectious bronchitis virus genomic RNA.

text: The subgenomic RNAs of the fowl coronavirus infectious bronchitis virus (IBV) form a 3' co-terminal or 'nested' set. The presence of non-contiguous (leader) sequences fused to the 5' termini of murine hepatitis virus mRNAs has been demonstrated using RNase T1 oligonucleotide mapping and sequencing. The presence of a leader sequence on IBV mRNA A has been demonstrated previously. In this paper the presence of a leader identical to that present on the 5' terminus of IBV mRNA A is demonstrated to be present on the 5' terminus of IBV genomic RNA. This has been achieved by sequencing of primer extension products and cDNA clones containing the genomic leader. Analysis of these clones has revealed the presence of a sequence at the leader/genome-length RNA junction which is closely related to regions of homology identified previously within the genomic RNA sequence at the leader/body junctions of subgenomic RNAs. The implications of this finding for mechanisms of coronavirus RNA synthesis are discussed.


title: Treatment of Chlamydia pneumoniae infection and chronic obstructive pulmonary disease.

text: Chronic obstructive lung disease (COPD) is a general term for chronic, irreversible lung disease that combines qualities of emphysema and chronic bronchitis. The standard definition of chronic bronchitis is a productive cough for three months per year (for at least two consecutive years) without an underlying aetiology. Acute exacerbation of chronic bronchitis (AECB) represents a common complaint that leads patients to seek medical attention. COPD and AECB are directly responsible for the overuse of antibiotics in the developed world. Fifty per cent of exacerbations have either viral or non-infectious origin. For this reason, antibiotic use remains controversial. Among other bacteria, Chlamydia pneumoniae is responsible for 4 - 16% of AECB in hospitalised or out-patients, although among smokers and people using steroids, the incidence is 34%. C. pneumoniae may either be the sole causative agent or a co-agent in AECB. This paper reviews the management of COPD/AECB with respect to antibiotic use. Diagnosis and antimicrobial therapy relevant to Chlamydia in the management of AECB are also evaluated in this review.


A virus-neutralising monoclonal antibody expressed in the milk of transgenic mice.


title: Severe acute respiratory syndrome.

text: PURPOSE OF REVIEW Severe acute respiratory syndrome (SARS) is an infectious disease first recognized in November 2002 in Guangdong Province, China. It spread to many countries all over the world during February to June 2003, with 8098 cases reported. Twenty-one percent of the affected people were health care workers. Because SARS is a new emerging disease, this review describes the current understanding about the etiology, clinical pictures, laboratory and radiological findings of SARS. RECENT FINDINGS Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) was quickly found to be the etiological agent of SARS in April 2003. The transmission of SARS-CoV between human beings is mainly due to close contact. Using barrier precautions, the transmission of SARS-CoV can be prevented. The most common clinical presentations of patients with SARS include fever, cough, and dyspnea. The common laboratory findings include lymphopenia, thrombocytopenia, elevated serum alanine and aspartate aminotransferase, lactate dehydrogenase, creatine phosphokinase, and C-reactive protein. The most common radiological finding is pneumonic lesion(s) in the chest radiogram. Many patients experience exacerbation of clinical symptoms in the second week of disease course and some may progress to respiratory failure and need mechanical ventilatory support. The overall case fatality rate is 9.6%. The current method of treatment of SARS is still controversial. SUMMARY SARS is an infectious disease with high contagiousness and a high mortality rate. Early case identification and infection control are two important factors to limit its spread.


The nature of viruses.


docid: xcrsbyot

title: An exploration of conceptual and temporal fallacies in international health law and promotion of global public health preparedness.

text: In February 2007, Indonesia withheld sharing H5N1 viral samples in order to compel the World Health Organization and Member States to guarantee future access to vaccines for States disproportionately burdened by infectious diseases. This article explores conceptual and temporal fallacies in the International Health Regulations (2005) and the Doha Declaration on the TRIPS Agreement and Public Health, as relates to global public health preparedness. Recommendations include adopting laws to facilitate non-pharmaceutical interventions; securing the rights of affected populations; and fostering inter-State collaborations to promote intra-State public health capacity building.


title: Molecular assays for detection of human metapneumovirus.

text: The recent description of the respiratory pathogen human metapneumovirus (hMPV) has highlighted a deficiency in current diagnostic techniques for viral agents associated with acute lower respiratory tract infections. We describe two novel approaches to the detection of viral RNA by use of reverse transcriptase PCR (RT-PCR). The PCR products were identified after capture onto a solid-phase medium by hybridization with a sequence-specific, biotinylated oligonucleotide probe. The assay was applied to the screening of 329 nasopharyngeal aspirates sampled from patients suffering from respiratory tract disease. These samples were negative for other common microbial causes of respiratory tract disease. We were able to detect hMPV sequences in 32 (9.7%) samples collected from Australian patients during 2001. To further reduce result turnaround times we designed a fluorogenic TaqMan oligoprobe and combined it with the existing primers for use on the LightCycler platform. The real-time RT-PCR proved to be highly reproducible and detected hMPV in an additional 6 out of 62 samples (9.6%) tested during the comparison of the two diagnostic approaches. We found the real-time RT-PCR to be the test of choice for future investigation of samples for hMPV due to its speed, reproducibility, specificity, and sensitivity.


title: Evaluation of the protection conferred by commercial vaccines against the California 99 isolate of infectious bronchitis virus.

text: An infectious bronchitis virus (IBV) was isolated from commercial broilers from the state of California exhibiting respiratory distress, inflamed tracheas, airsaculitis, and edematous lungs. After reverse transcriptase-polymerase chain reaction (RT-PCR), the California isolate exhibited an identical restriction fragment length polymorphism (RFLP) pattern to some isolates obtained from California, known as California 99 isolates. Commercial Mass-Conn and Mass-Ark vaccines were used to vaccinate commercial broiler chickens via eye drop once at 1 or 10 days of age or twice at 1 and 10 days of age. At 27 days of age the birds were challenged via eye drop with the isolated IBV California 99 strain. Protection was measured by failure to reisolate the challenge virus from tracheas 5 days postchallenge and complemented withthe tracheal and epithelium thickness scores. When the Mass-Ark vaccine was included in the vaccination programs, there was protection against challenge with the IBV California 99 isolate. The Mass-Conn vaccine conferred protection when used once at 1 day of age and twice at 1 and 10 days of age. However, no total protection was achieved when used as the only vaccine at 10 days of age, since one of the replicates was positive for virus isolation. Significant differences (P < 0.05) in the epithelium thickness and tracheal scores were observed between the unvaccinated-unchallenged group and the groups vaccinated once or twice with the Mass-Conn vaccine. Based on these results, all chickens were protected against the California 99 isolate when the IBV Arkansas type was used as a vaccine.


title: S1 of distinct IBV population expressed from recombinant adenovirus confers protection against challenge.

text: Protective properties of three distinct infectious bronchitis virus (IBV) Ark Delmarva poultry industry (ArkDPI) S1 proteins encoded from replication-defective recombinant adenovirus vectors were investigated. Using a suboptimal dose of each recombinant virus, we demonstrated that IBV S1 amino acid sequences showing > or = 95.8% amino acid identity to the S1 of the challenge strain differed in their ability at conferring protection. Indeed, the S1 sequence of the IBV population previously designated C4 (AdIBVS1.C4), which protected the most poorly, differs from the S1 sequence of population C2 (AdIBVS1.C2), which provided the highest protection, only at amino acid position 56. The fact that a change in one amino acid in this region significantly altered the induction of a protective immune response against this protein provides evidence that the first portion of S1 displays relevant immunoprotective epitopes. Use of an optimal dose of AdIBVS1.C2 effectively protected chickens from clinical signs and significantly reduced viral load after IBV Ark virulent challenge. Moreover, increased numbers of both IgA and IgG IBV-specific antibody secreting lymphocytes were detected in the spleen after challenge. The increased response detected for both IgA and IgG lymphocytes after challenge might be explained by vaccine-induced B memory cells. The fact that a single vaccination with Ad/IBVS1.C2 provides protection against IBV challenge is promising, because Ad-vectored vaccines can be mass delivered by in ovo inoculation using automated in ovo injectors.


title: Comparison of expression patterns and cell adhesion properties of the mouse biliary glycoproteins Bbgp1 and Bbgp2.

text: Biliary glycoproteins are members of the carcinoembryonic antigen (CEA) family and behave as cell adhesion molecules. The mouse genome contains two very similar Bgp genes, Bgp1 and Bgp2, whereas the human and rat genomes contain only one BGP gene. A Bgp2 isoform was previously identified as an alternative receptor for the mouse coronavirus mouse hepatitis virus. This isoform consists of two extracellular immunoglobulin domains, a transmembrane domain and a cytoplasmic tail of five amino acids. In this report, we have examined whether the Bgp2 gene can express other isoforms in different mouse tissues. We found only one other isoform, which has a long cytoplasmic tail of 73 amino acids. The long cytodomain of the Bgp2 protein is highly similar to that of the Bgp1/4L isoform. The Bgp2 protein is expressed in low amounts in kidney and in a rectal carcinoma cell line. Antibodies specific to Bgp2 detected a 42-kDa protein, which is expressed at the cell surface of these samples. Bgp2 was found by immunocytochemistry in smooth muscle layers of the kidney, the uterus, in gut mononuclear cells and in the crypt epithelia of intestinal tissues. Transfection studies showed that, in contrast with Bgp1, the Bgp2 glycoprotein was not directly involved in intercellular adhesion. However, this protein is found in the proliferative compartment of the intestinal crypts and in cells involved in immune recognition. This suggests that the Bgp2 protein represents a distinctive member of the CEA family; its unusual expression patterns in mouse tissues and the unique functions it may be fulfilling may provide novel clues about the multiple functions mediated by a common BGP protein in humans and rats.


title: Completion of the sequence of the genome of the coronavirus avian infectious bronchitis virus.

text: The nucleotide sequence determination of the genome of the Beaudette strain of the coronavirus avian infectious bronchitis virus (IBV) has been completed. The complete sequence has been obtained from 17 overlapping cDNA clones, the 5'-most of which contains the leader sequence (as determined by direct sequencing of the genome) and the 3'-most of which contains the poly(A) tail. Approximately 8 kilobases at the 3' end of this sequence have already been published. These contain the sequences of mRNAs A to E within which are the genes for the spike, the membrane and the nucleocapsid polypeptides: the main structural components of the virion. The remainder of the sequence, equivalent to the 'unique' region of mRNA F, is some 20 kilobases in length and is thought to code for a polymerase or polymerases which are involved in the replication of the genome and the production of the subgenomic messenger RNAs. This sequence contains two large open reading frames, potentially coding for polypeptides of molecular weights 441,000 and 300,000. Unlike other large open reading frames in the virus, the 300,000 open reading frame appears to have no subgenomic RNA associated with it which would allow it to be at the 5' end of an mRNA species. Because of this, and because of the characteristics of the sequence in the region immediately upstream of its start codon, other mechanisms of translation, such as ribosome slippage, must be postulated.


title: Prevalence of bovine group A rotavirus shedding among dairy calves in Ohio.

text: Fecal samples were collected from 450 neonatal calves, ranging from 1 to 30 days old, between May, 1988 and May, 1989 to estimate the prevalence of bovine group A rotavirus in a stratified random sample of Ohio dairy herds. Calves were from 47 dairy herds chosen to be representative of Ohio herds. Bovine group A rotavirus was detected in fecal samples by a cell culture immunofluorescence test (CCIF) and ELISA. Of 450 samples tested, 46 (10%) were positive by CCIF and 67 (15%) were positive by ELISA. The agreement beyond chance between the 2 assays was good (kappa = 0.65). The overall prevalence rate of rotavirus shedding was 16.4% (74/450). Forty-three percent (29/67) of the samples positive by ELISA were subgroup 1, none were subgroup 2, and the remaining 57% (38/67) could not be assigned to either subgroups 1 or 2. Thirty herds (62.5%) had at least 1 group A rotavirus-positive calf (mean number of samples per positive herd = 12.4), and 17 herds (37.5%) had no rotavirus-positive calves (mean number of samples per negative herd = 6.0). A live oral rotacoronavirus vaccine was used in neonatal calves of only 1 herd and 3 of 17 (17.6%) calves from this herd were positive for group A rotavirus. The percentage of the rotavirus-positive fecal samples from all calves (n = 450) when stratified by fecal consistency was as follows: 28.3% (13/46) had liquid feces; 25.6% (10/39) had semiliquid feces; 23.4% (22/94) had pasty feces; and 10.7% (29/271) had firm feces.(ABSTRACT TRUNCATED AT 250 WORDS)


title: Endovascular clot retrieval for acute ischaemic stroke: the Auckland City Hospital experience.

text: AIMS In acute ischaemic stroke, endovascular therapy with the Solitaire FR stent retriever has been shown to double recanalisation rates and the numbers of patients who recover to be functionally independent, when compared to standard therapy. We present the Auckland City Hospital experience of clot retrieval. METHODS Previously independent ischaemic stroke patients with contraindications to, or no response following, i.v. alteplase, were treated with clot retrieval. All patients had proximal large artery occlusions on CT angiography and many also had CT perfusion scans showing salvageable ischaemic tissue. RESULTS Clot retrieval was performed in 33 patients (10 women, mean (SD) age of 54 (17) years) since 2011. Twenty-two (67%) patients were first treated with alteplase. Patients fell into three groups: 17 (52%) had anterior circulation occlusion, similar to those in recent clot retrieval studies; 10 (30%) had posterior circulation occlusion; and six (18%) had 'Rescue' clot retrieval, usually with stroke that followed a procedure. Patients with anterior circulation occlusion had a median time from symptom onset to groin puncture of 225 (range 95-450) minutes, full recanalisation occurred in 76%, and by day 90, all 17 were alive and living at home, with 63% functionally independent (modified Rankin Scale (mRS) 0-2). At day 90, eight of 10 posterior circulation occlusion group patients were alive and living at home, four with a mRS of 0-2. In contrast, four of six 'Rescue' patients had died, and another was functionally dependent with a mRS of 4. CONCLUSIONS Endovascular clot retrieval can be safely and effectively performed in a New Zealand setting with similar results to recent trials in anterior circulation occlusion patients. We suggest that District Health Boards develop clot retrieval services as part of regional hyperacute stroke treatment pathways.


title: Expression of MHC class I genes in mouse hepatitis virus (MHV-A59) infection and in multiple sclerosis.

text: Our studies revealed that virus induced demyelination as well as human inflammatory demyelination involves upregulation of class I MHC genes and surface expression of antigens encoded by these genes. Induction involves the action of an intermediate soluble factor/s which is at present unknown. These findings suggest that MHC class I restricted, cytotoxic T lymphocyte (CTL) reactions, against self or foreign antigens may play a role in these conditions. These findings may help to elucidate the mechanism of coronavirus-induced demyelination as well as the pathogenesis of multiple sclerosis.


docid: amxrgvhv

title: [Dynamics of peripheral blood lymphocytes and their subpopulations in patients with severe acute respiratory syndrome].

text: OBJECTIVE To study on the dynamics of peripheral blood lymphocytes and their subpopulations in patients with severe acute respiratory syndrome. METHODS Using flow cytometry, the absolute numbers of peripheral blood lymphocytes and their subpopulations in 240 SARS patients (696 specimens) and 51 individuals as controls, were counted and compared. RESULTS The absolute numbers of peripheral blood lymphocytes and their subpopulations (CD45, CD3, CD4, CD8) were 1298 +/- 785, 897 +/- 606, 510 +/- 372, 362 +/- 263/mm(3), respectively, significantly lower in SARS patients as compared to the normal controls (2024 +/- 423, 1391 +/- 289, 795 +/- 129, 551 +/- 183/mm(3)). Of SARS patients, severe group (1095 +/- 740, 740 +/- 562, 419 +/- 346, 304 +/- 244/mm(3)) had lower counts than that of mild group (1404 +/- 788, 991 +/- 612, 564 +/- 378, 396 +/- 267/mm(3)), and in group with deaths (587 +/- 493, 369 +/- 371, 204 +/- 191, 150 +/- 130/mm(3)) was lower than that of recovery group (1355 +/- 776, 948 +/- 603, 539 +/- 375, 382 +/- 263/mm(3)). There were significant differences (P < 0.01) for CD45, CD3, CD4, CD8, but with no significant difference (P > 0.05) for CD4/CD8 ratio between severe and mild, recovery and death groups. The lymphocytes and their subpopulations (CD45, CD3, CD4, CD8) declined in the 1st week and to the lowest level (977 +/- 579, 641 +/- 466, 360 +/- 275, 270 +/- 216/mm(3)) in the 2nd week. Then the lymphocytes and their subpopulations gradually increased during the recovery of the disease. CONCLUSION The absolute numbers of peripheral blood lymphocytes and their subpopulations in SARS patients might be used as one of the methods for diagnosis on the severity and prognosis of the disease.


title: The nsp2 replicase proteins of murine hepatitis virus and severe acute respiratory syndrome coronavirus are dispensable for viral replication.

text: The positive-stranded RNA genome of the coronaviruses is translated from ORF1 to yield polyproteins that are proteolytically processed into intermediate and mature nonstructural proteins (nsps). Murine hepatitis virus (MHV) and severe acute respiratory syndrome coronavirus (SARS-CoV) polyproteins incorporate 16 protein domains (nsps), with nsp1 and nsp2 being the most variable among the coronaviruses and having no experimentally confirmed or predicted functions in replication. To determine if nsp2 is essential for viral replication, MHV and SARS-CoV genome RNA was generated with deletions of the nsp2 coding sequence (MHVDeltansp2 and SARSDeltansp2, respectively). Infectious MHVDeltansp2 and SARSDeltansp2 viruses recovered from electroporated cells had 0.5 to 1 log10 reductions in peak titers in single-cycle growth assays, as well as a reduction in viral RNA synthesis that was not specific for any positive-stranded RNA species. The Deltansp2 mutant viruses lacked expression of both nsp2 and an nsp2-nsp3 precursor, but cleaved the engineered chimeric nsp1-nsp3 cleavage site as efficiently as the native nsp1-nsp2 cleavage site. Replication complexes in MHVDeltansp2-infected cells lacked nsp2 but were morphologically indistinguishable from those of wild-type MHV by immunofluorescence. nsp2 expressed in cells by stable retroviral transduction was specifically recruited to viral replication complexes upon infection with MHVDeltansp2. These results demonstrate that while nsp2 of MHV and SARS-CoV is dispensable for viral replication in cell culture, deletion of the nsp2 coding sequence attenuates viral growth and RNA synthesis. These findings also provide a system for the study of determinants of nsp targeting and function.


title: Intraoperative pediatric blood transfusion therapy: a review of common issues. Part I: hematologic and physiologic differences from adults; metabolic and infectious risks.

text: Pediatric intraoperative transfusion therapy, particularly the approach to massive blood transfusion (blood loss > or =one blood volume) can be quite complex because of the unique relationship between the patient's blood volume and the volume of the individual blood product transfused. This paper is divided into two parts: part 1 presents an overview of the physiologic and hematologic differences between children and adults as well as an overview of the metabolic consequences of blood transfusions, risks of disease transmission, and blood compatibility issues.


title: Emergency Colorectal Surgery in a COVID-19 Pandemic Epicenter.

text: Transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a considerable risk during emergency colorectal surgery in a pandemic epicenter. It is well known that the primary route of SARS-CoV-2 transmission is through respiratory droplets. However, little is known about shedding of the virus in bodily fluids and associated risks. Although the current moratorium on elective surgery addresses multiple ongoing concerns, including the management of precious resources as well as unknown exposure risks, surgeons undeniably must face and mitigate risks related to exposure to patient airway management-related aerosols, bodily fluids, surgical smoke, contaminated insufflation, and specimen handling in emergency colorectal surgery. Given the significant concern of airborne transmission, the authors recommend conventional, in lieu of laparoscopic, access in emergency colorectal surgery in a COVID-19 pandemic epicenter.


Screening for Barrett Esophagus With a Nonendoscopic Sponge Capsule.


title: The public's response to severe acute respiratory syndrome in Toronto and the United States.

text: Using data from 13 surveys of the public, this article compares the public's response to severe acute respiratory syndrome (SARS) in Ontario (specifically, Toronto), the other Canadian provinces, and the United States, which had substantial differences in the number of SARS cases. Findings suggest that, even at a relatively low level of spread among the population, the SARS outbreak had a significant psychological and economic impact. They also suggest that the success of efforts to educate the public about the risk of SARS and appropriate precautions was mixed. Some of the community-wide problems with SARS might have been avoided with better communication by public health officials and clinicians.


Using family visitors, sitters, or volunteers to prevent inpatient falls.


title: Processing and subcellular localization of the leader papain-like proteinase of Beet yellows closterovirus.

text: ORF 1a of Beet yellows closterovirus (BYV) encodes the domains of the papain-like proteinase (PCP), methyltransferase (MT) and RNA helicase. BYV cDNA inserts encoding the PCP-MT region were cloned in pGEX vectors next to the glutathione S-transferase gene (GST). In a 'double tag' construct, the GST-PCP-MT cDNA was flanked by the 3'-terminal six histidine triplets. Following expression in E. coli, the fusion proteins were specifically self-cleaved into the GST-PCP and MT fragments. MT-His(6) was purified on Ni-NTA agarose and its N-terminal sequence determined by Edman degradation as GVEEEA, thus providing direct evidence for the Gly(588)/Gly(589) bond cleavage. The GST-PCP fragment purified on glutathione S-agarose was used as an immunogen to produce anti-PCP monoclonal antibodies (mAbs). On Western blots of proteins from virus-infected Tetragonia expansa, the mAbs recognized the 66 kDa protein. Immunogold labelling of BYV-infected tissue clearly indicated association of the PCP with the BYV-induced membranous vesicle aggregates, structures related to closterovirus replication.


title: Molecular detection and serotyping of infectious bronchitis virus from FTA filter paper.

text: We investigated the feasibility of using Flinders Technology Associates (FTA) filter cards for the storage of allantoic fluid containing an infectious bronchitis virus (IBV), such as Arkansas-DPI, Connecticut, and Massachusetts, and for their identification by reverse transcriptase (RT)-polymerase chain reaction (PCR) and characterization by restriction fragment length polymorphism (RFLP) or nucleotide sequencing. FTA paper is a cotton-based cellulose membrane containing lyophilized chemicals that lyses many types of bacteria and viruses. IBV was inactivated upon contact with the FTA, as shown by the inability of the virus to be propagated in embryonating chicken eggs. RT-PCR of the S1 gene showed that viral RNA in allantoic fluid remained stable after storage on FTA filter cards and that the stability was time and temperature sensitive for the large (1700 base pair [bp]) but not the small (383 bp) PCR products. Analysis of the amplified products showed that molecular characterization is feasible in allantoic fluid stored on FTA under nonfavorable environmental conditions (41 C) for at least 15 days. The use of FTA cards for the collection, transport, and storage of IBV-containing samples is safe, inexpensive, and adequate for molecular diagnosis. We propose that specimens coming from overseas on FTA cards would be first analyzed by RT-PCR with primers yielding a 1700-bp product followed by RFLP of the positive cases. Negative cases would be analyzed with primers yielding a 383-bp product (to exdude detrimental effect of the storage conditions) followed by nucleotide sequencing of the positive cases.


Intussusception of the spiral colon associated with fibroserous granulation in a heifer.


title: [Follow up study on viruses associated with SARS among the SARS patients].

text: BACKGROUND To study the existence status of the SARS-CoV, retrovirus, and the poliovirus in the bodies of the patients with SARS and the possible relationship between the three viruses and SARS. METHODS The clinical specimens of the nasopharyngeal swabs, sputum (or saliva), urine, fecal specimens were collected on three consecutive days from 8 patients with SARS 2 years after the recovery from SARS. SARS-CoV, reovirus and poliovirus RNA was detected by using reverse transcription (RT)-PCR; IgG antibody to the poliovirus type 1 and 3 and the antibody to SARS-CoV were determined using enzyme linked immunosorbent assay (ELISA). RESULTS All the specimens were negative for SARS-CoV and reovirus by RT-PCR, but the fecal specimens from 4 persons were positive for poliovirus. The sequences of these poliovirus were highly homologous to that of human poliovirus type 1 strain sabin 1 genome at nucleotide level, but back mutations have occurred in the primary attenuating mutation sites at nucleotide position 480 (G --> A) in the 5' UTR and the nucleotide position 2795 (A --> G). No SARS-CoV, reovirus, and poliovirus were found in the normal controls. Three serum specimens were positive for the antibody to SARS-CoV. The IgG antibody to poliovirus were detected in 4 SARS patients and 23 healthy persons. No positive results for antibody to SARS-CoV were detected in the 25 healthy persons. CONCLUSION The positive rate of the poliovirus antibody in the serum of SARS patients 2 years after recovery was significantly different from that of the normal controls, and the positive rate of poliovirus in the fecal specimens was still very high, and more importantly back mutations have occurred in the attenuating mutation sites at nucleotide position which plays an important role in the poliomyelitis.


title: Putative hAPN receptor binding sites in SARS_CoV spike protein.

text: AIM To obtain the information of ligand-receptor binding between the S protein of SARS-CoV and CD13, identify the possible interacting domains or motifs related to binding sites, and provide clues for studying the functions of SARS proteins and designing anti-SARS drugs and vaccines. METHODS On the basis of comparative genomics, the homology search, phylogenetic analyses, and multi-sequence alignment were used to predict CD13 related interacting domains and binding sites in the S protein of SARS-CoV. Molecular modeling and docking simulation methods were employed to address the interaction feature between CD13 and S protein of SARS-CoV in validating the bioinformatics predictions. RESULTS Possible binding sites in the SARS-CoV S protein to CD13 have been mapped out by using bioinformatics analysis tools. The binding for one protein-protein interaction pair (D757-R761 motif of the SARS-CoV S protein to P585-A653 domain of CD13) has been simulated by molecular modeling and docking simulation methods. CONCLUSION CD13 may be a possible receptor of the SARS-CoV S protein, which may be associated with the SARS infection. This study also provides a possible strategy for mapping the possible binding receptors of the proteins in a genome.


title: Severe acute respiratory syndrome: prognostic implications of chest radiographic findings in 52 patients.

text: PURPOSE To retrospectively assess prognostic implications of radiographic findings in severe acute respiratory syndrome (SARS). MATERIALS AND METHODS Radiographic findings were reviewed by two radiologists for 52 patients with SARS. On each radiograph, each lung was separated into upper, middle, and lower zones. A four-point scale was used to score extent of SARS-related lesions in each zone; points from all zones were added for a cumulative score. Patient sex, age, comorbidities, duration of developing lesions, lesion score for each radiograph, need for mechanical ventilation, and percentage of lung affected were compared between patients who died (n = 20) and survivors (n = 32). Continuous and categorical variables were analyzed with Mann-Whitney test and Fisher exact or chi(2) test, respectively. RESULTS Survival and mortality groups showed no significant differences with respect to patient sex, duration of SARS-related lesions, development of lesion shifting, and acute respiratory distress syndrome. Patients who died were significantly older (mean +/- standard deviation, 56.9 years +/- 17.2 vs 40.4 years +/- 16.6; P =.002) and had higher frequency of comorbid lung illnesses (nine of 20 vs two of 32, P =.001), maximal lesion extent score of 7 or higher (20 of 20 vs five of 32, P <.001), involvement of four or more lung zones (17 of 20 vs four of 32, P <.001), bilateral lung involvement (19 of 20 vs 14 of 32, P <.001), need for mechanical ventilation (18 of 20 vs two of 32, P <.001), and higher percentage of affected areas (41.5% +/- 8.6 vs 16.4% +/- 10.0, P <.001) than those of survivors. CONCLUSION On chest radiographs, maximal SARS-related lesion extent score of 7 or higher is a strong predictor of mortality, especially in patients with comorbid lung illnesses and involvement of four or more lung zones.


title: SARS: understanding the virus and development of rational therapy.

text: In late 2002 a new disease, severe atypical respiratory syndrome (SARS), emerged in China. A hitherto unknown animal coronavirus (CoV) that had crossed the species barrier through close contact of humans with infected animals was identified as the etiological agent. It rapidly adapted to the new host and not only became readily transmissible between humans but also more pathogenic. Air travel spread it rapidly around the world and ultimately the virus infected 8096 people and caused 774 deaths in 26 countries on 5 continents. Aggressive quarantine measures successfully terminated the disease. Currently, there are no SARS cases recorded and most likely the virus no longer circulates in the human population. In this review we present an overview over SARS-Co virus biology, the disease and discuss strategies to develop antiviral drugs and vaccines.


docid: uecqvrjp

title: Togaviridae.

text: The family Togaviridae comprises four genera: Alphavirus (with 26 species), Rubivirus (one species), Pestivirus (three species), and Arterivirus (one species). The main characteristics of the member viruses are: (i) the virus particles are spherical, 50-70 nm in diameter, including an envelope with surface projections that incorporate two or three polypeptides, usually glycosylated; (ii) the nucleocapsid comprises a core protein and a single strand of positive-sense RNA, molecular weight about 4 X 10(6); where characterized, the RNA has an m7G 'cap' at the 5' end and is polyadenylated at the 3' end; (iii) maturation occurs by budding of spherical nucleocapsids 30-35 nm in diameter, with proven or presumed icosahedral symmetry, through cytoplasmic membranes. Where characterized, translation of structural proteins occurs on subgenomic messenger RNA(s); these appear to represent the 3' end of the genome. Nearly all alphavirus species are transmitted by mosquitoes. Transmission also occurs transovarially (Alphavirus) or transplacentally (Rubivirus and Pestivirus). Members of a genus are serologically related, but are not related to members of other genera.


title: Togaviridae.

text: The family Togaviridae is described; it contains four genera--Alphavirus, Flavivirus, Pestivirus and Rubivirus--and additional members. The main characteristics of the family are as follows: single-stranded, linear RNA, molecular weight about 4 X 10(6). Virions have isometric nucleocapsids surrounded by a lipoprotein envelope containing host cell lipid and virus-specified polypeptides including one or more glycopeptides. Virions yield infectious RNA. There are at least 80 members; all the alphaviruses and most flaviviruses are arboviruses in the biological sense.


docid: wpcdpycq

title: The psychosocial effects of being quarantined following exposure to SARS: a qualitative study of Toronto health care workers.

text: OBJECTIVE To examine the psychosocial effects on health care workers of being quarantined because of exposure to severe acute respiratory syndrome (SARS). METHOD We used semistructured qualitative interviews. RESULTS We identified 3 major themes concerning psychosocial effects: loss, duty, and conflict. CONCLUSIONS Quarantined workers experienced stigma, fear, and frustration. We highlight the need for clear and easily accessible information on dealing with infectious diseases. Practical advice on coping and stress management techniques for health care workers are needed in preparation for potential future outbreaks of infectious diseases.


title: Diagnostic value of real-time capillary thermal cycler in virus detection.

text: Molecular-based detection methods such as PCR techniques have had a significant impact on the diagnosis of viral infections because of their superior sensitivity and rapid turnaround time. This review describes the use of real-time PCR on the capillary thermal cycler, the Roche LightCycler trade mark, for early disease detection in diagnostic virology. The advantages of using the LightCycler, the detection processes using SYBR Green I and different hybridization strategies will be discussed in detail, with specific examples drawn from our in-house viral assays. The use of the LightCycler for the investigation of two recent viral outbreaks in Singapore will also be briefly described.


Analysis of nonessential gene function in recombinant MHV-JHM. Gene 4 knockout recombinant virus.


title: Osteomyelitis and Discitis Following Translumbar Repair of a Type II Endoleak.

text: Here we present the case of an 80-year-old man who developed a type II endoleak following endovascular abdominal aortic aneurysm repair. Initial attempts at treating the endoleak via a transarterial approach were unsuccessful; therefore the patient underwent percutaneous translumbar endoleak embolization. Approximately 1 month following the translumbar procedure, he developed back pain, with subsequent workup revealing osteomyelitis and discitis as a complication following repair via the translumbar approach.


title: Preliminary observations on enteritis associated with a coronavirus-like agent in rabbits.

text: An outbreak of fatal enteritis in 3-8-week old rabbits in a barrier-maintained breeding colony is described. The laboratory findings and treatments attempted suggest that a viral agent was the primary pathogen.


The Ministry of Health.


title: Reverse genetic analysis of the transcription regulatory sequence of the coronavirus transmissible gastroenteritis virus.

text: Coronavirus discontinuous transcription uses a highly conserved sequence (CS) in the joining of leader and body RNAs. Using a full-length infectious construct of transmissable gastroenteritis virus, the present study demonstrates that subgenomic transcription is heavily influenced by upstream flanking sequences and supports a mechanism of transcription attenuation that is regulated in part by a larger domain composed of primarily upstream flanking sequences which select appropriately positioned CS elements for synthesis of subgenomic RNAs.


Background paper. The appearance of the porcine respiratory coronavirus has created new problems and perspectives.


docid: pibmnbrx

title: Preparing the biochemistry laboratory for the next outbreak: lessons from SARS in Singapore.

text: Severe acute respiratory syndrome (SARS) is an emerging disease characterised by fever and atypical pneumonia and caused by a novel coronavirus. Singapore was affected by the global pandemic in early 2003, with 238 cases and 33 deaths. Samples sent to the biochemistry laboratory made up the majority (69%) of all SARS samples, yet remained a minority (29%) of total biochemistry workload. This paper describes the problems encountered and solutions adopted by the biochemistry laboratory at the designated SARS hospital in coping with this epidemic. It provides practical advice for laboratories planning for the handling of samples from future outbreaks.


docid: iugbpcrt

title: [A new method for visual SARS DNA sequences analysis].

text: Traditional DNA sequence analysis is based on sequence alignment, while a new DNA visual sequence analysis is proposed in this paper. Based on S. Wolfram's cellular automation theory, the method transfers one-dimensional DNA sequence into two-demensional visual image. Applying this method to SARS DNA sequence analysis, a characteristic of SARS-CoV differing from non-SARS is discovered. Compared with all known coronaviruses' images, It is found that this is a unique characteristic of SARS virus, and it is helpful to clinical identification of SARS.


title: Suppression of acute anti-friend virus CD8+ T-cell responses by coinfection with lactate dehydrogenase-elevating virus.

text: Friend virus (FV) and lactate dehydrogenase-elevating virus (LDV) are endemic mouse viruses that can cause long-term chronic infections in mice. We found that numerous mouse-passaged FV isolates also contained LDV and that coinfection with LDV delayed FV-specific CD8(+) T-cell responses during acute infection. While LDV did not alter the type of acute pathology induced by FV, which was severe splenomegaly caused by erythroproliferation, the immunosuppression mediated by LDV increased both the severity and the duration of FV infection. Compared to mice infected with FV alone, those coinfected with both FV and LDV had delayed CD8(+) T-cell responses, as measured by FV-specific tetramers. This delayed response accounted for the prolonged and exacerbated acute phase of FV infection. Suppression of FV-specific CD8(+) T-cell responses occurred not only in mice infected concomitantly with LDV but also in mice chronically infected with LDV 8 weeks prior to infection with FV. The LDV-induced suppression was not mediated by T regulatory cells, and no inhibition of the CD4(+) T-cell or antibody responses was observed. Considering that most human adults are carriers of chronically infectious viruses at the time of new virus insults and that coinfections with viruses such as human immunodeficiency virus and hepatitis C virus are currently epidemic, it is of great interest to determine how infection with one virus may impact host responses to a second infection. Coinfection of mice with LDV and FV provides a well-defined, natural host model for such studies.


title: Does Topical Lidocaine Reduce the Pain Associated With the Insertion of Nasal Continuous Positive Airway Pressure Prongs in Preterm Infants?: A Randomized, Controlled Pilot Trial.

text: OBJECTIVE To evaluate the efficacy of topical lidocaine 2% gel in reducing the pain associated with the insertion of nasal continuous positive airway pressure (nCPAP) prongs in preterm infants. MATERIALS AND METHODS A pilot randomized controlled trial. Sixty preterm infants, categorized into lidocaine (n=30) and control groups (n=30). The primary outcome was Premature Infant Pain Profile (PIPP) score, secondary outcomes included salivary cortisol, presence of cry, the duration of first cry, and adverse effects of lidocaine. RESULTS There were no statistically significant differences between lidocaine and control groups regarding PIPP scores (mean±SD: 7.2±2.3 vs. 9.3±3.0, respectively, P=0.086). None of the infants in the lidocaine group had severe pain defined as a PIPP score>12, compared with 3 (10%) infants in the control group (P=0.056). Salivary cortisol concentrations were not significantly different between the lidocaine and control groups (mean±SD: 2.57±1.79 vs. 4.82±1.61 μg/dL, respectively, P=0.11). Standardized effect sizes for topical lidocaine were medium to large for reduction in PIPP scores and large for reduction in salivary cortisol (Cohen d=-0.78 and -1.32, respectively). No adverse effects were reported in infants receiving lidocaine. DISCUSSION Our data suggest that topical lidocaine did not reduce the pain associated with the insertion of nCPAP prongs in preterm infants. However, the trends for lower PIPP scores in the lidocaine group and the effect sizes for lidocaine on PIPP scores and salivary cortisol were large enough so that a large-scale randomized clinical trial is warranted to confirm or refute our results. Such a study should compare 2 or more active pain interventions during nCPAP application, rather than evaluating a single intervention versus placebo or no treatment.


title: Redistributing work in aging Europe.

text: As Europe ages, the proportion of people who work will decline unless older individuals remain in the labor force. Such reform could be part of a more general redistribution of work. If a greater share of the population worked, then the average number of hours worked per week could be reduced. This could particularly help younger people and increase Europe's low birth rates. The challenges facing Germany, Europe's most populous country, are highlighted, but statistics are also given for five other European countries and, for comparison, the United States. Social science research is needed to provide policy-relevant knowledge about life-course options.


title: Knowledge of and attitudes toward severe acute respiratory syndrome among a cohort of dental patients in Hong Kong following a major local outbreak.

text: OBJECTIVE To assess the knowledge of and attitudes toward severe acute respiratory syndrome (SARS) among patients attending a teaching dental hospital and private dental practices in Hong Kong during a major local SARS outbreak. METHODS 250 dental patients were interviewed by questionnaire and 213 were interviewed by phone. RESULTS Less than one-third (30.0%) of the 463 respondents said they were not afraid of contracting the SARS coronavirus from their dentists and did not avoid dental treatment for that reason. Nearly three-fifths (56.7%) did not worry about contracting SARS from dental treatment. Fewer than 10% of the respondents thought that dentists ran a high risk of contracting SARS. From the patients' experiences, 85.2% and 21.7% of the dentists wore face masks and face shields, respectively, when delivering dental treatment. CONCLUSIONS The majority of patients interviewed had confidence in their dentists, their treatment environments, and the infection control measures taken, and were not worried about contracting SARS in the dental setting. This perception is an improvement from that described in an earlier study, in which more than half of the patients were concerned about contracting an infection during dental treatment and perceived that the infection control measures undertaken by the dental profession to prevent infectious diseases were not satisfactory. Patients, however, demanded better infection control measures during the SARS outbreak.


title: Impacting Canadian public health nurses' job satisfaction.

text: OBJECTIVE Workforce recruitment and retention challenges are being experienced in public health as in other Canadian health sectors. While there are many nurses working in public health, little research has been done about their job satisfaction. Job satisfaction is linked to recruitment, retention and positive client outcomes. The purpose of this research was to examine the relationships between three modifiable work environment factors (autonomy, control-over-practice, and workload) and Canadian public health nurses' (PHNs) job satisfaction. METHODS Data were from the 2005 National Survey of the Work and Health of Nurses (response rate, 79.7%; 18,676 nurses). Bivariate and multivariate logistic regression analyses were used for this secondary analysis. Findings were discussed with practicing PHNs, policy-makers and researchers from across Canada at a knowledge translation (KT) 'Think-Tank'. RESULTS Among the 271 PHNs, 53.5% reported being 'very satisfied' with their jobs. The interaction between autonomy and workload was a significant predictor of PHNs' job satisfaction, (OR 0.97, 95% CI 0.96-0.99, p < 0.01) as was the interaction between age and workload (OR 1.01, 95% CI 1.00-1.01, p < 0.01). Think-Tank participants selected priority areas for application to public health practice, management and research. CONCLUSION Despite being an important practice factor, this is the first study to reveal the negative influence of PHNs' autonomy when in interaction with an excessive workload. Significant workload findings and the presence of generational differences suggest the need for development of workload measurement tools and public health human resource strategies tailored to a multi-generational workforce.


title: [Detection and analysis of SARS coronavirus-specific antibodies in sera from non-SARS children].

text: OBJECTIVE To examine the presence of severe acute respiratory syndrome (SARS) coronavirus-specific antibodies in the sera from non-SARS children. METHODS Indirect immunofluorescent assay and double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) were used to detect the virus-specific antibodies in sera of 1,060 non-SARS children in Guangzhou. RESULTS All the serum samples from the 1,060 non-SARS children were negative for both IgG and IgM antibodies against SARS coronavirus as determined by indirect immunofluorescent assay, with only two serum samples showing weak positivity for SARS coronavirus-specific antibodies identified by double-antigen sandwich ELISA. CONCLUSION No SARS coronavirus-specific antibody are present in the sera of non-SARS children.


Global Fund withdraws grants to Myanmar.


title: Studies on the hemagglutination phenomenon of hemagglutinating encephalomyelitis virus (HEV) of pigs.

text: Hemagglutinating encephalomyelitis virus (HEV), unlike other Coronaviruses, readily agglutinates a variety of red blood cells. The reaction differs from that of the Myxovirus/Paramyxovirus groups in several respects. Unlike the Myxoviruses, HEV agglutinated red cells treated with receptor destroying enzyme. No neuraminidase activity was demonstrated with HEV and there was no apparent elution of the virus from agglutinated cells. Mucins and animal sera inhibited the agglutination reaction markedly. Ultra violet light exposure sufficient to destroy infectivity had no effect on hemagglutination. Density gradient studies showed that the hemagglutinin was closely associated with the virus particle.


title: Enzyme-linked immunosorbent assay reactivity of torovirus-like particles in fecal specimens from humans with diarrhea.

text: Toroviruses are recognized enteric pathogens of cattle and horses; in humans, similar pleomorphic particles have been described, but doubt has been raised concerning their identity as viruses. We screened fecal samples from humans with diarrhea for the presence of torovirus-like particles (TVLPs) by electron microscopy and subsequently used an enzyme-linked immunosorbent assay (ELISA) with bovine torovirus reference reagents to test for the presence of torovirus antigens. To add another selection criterion to this heterologous ELISA, we enriched the TVLPs from the stool specimens by using sucrose density gradients before testing. The results of ELISA and EM correlated significantly, the ELISA having a sensitivity of 68% and a specificity of 86% (chi-square, P < 0.0001). In the gradient, peaks of ELISA reactivity were found at a buoyant density of 1.16 g/ml and were parallel to those found when using bovine torovirus. Furthermore, in 50% of the ELISA-positive gradients, a hemagglutinin for human group O erythrocytes comigrated with the peaks of ELISA reactivity. We were unable to isolate human TVLPs in human colonic tumor or rectal tumor cells. We cloned and sequenced amplification products obtained by low-stringency polymerase chain reaction amplification using consensus primers mapping to the 3' end of the genome of animal toroviruses, but found no significant homologies with animals torovirus sequences. Rabbits were inoculated with material from the gradient peak fractions of human stool specimens, and their sera were assayed for immunologic comparison with bovine torovirus as a reference. A two-way antigenic cross-reactivity was seen between human TVLP and bovine torovirus reagents when tested by ELISA. The rabbit antisera to human TVLP detected a higher number of electron microscopy-positive stool specimens than did the rabbit antisera to bovine torovirus. The application of these assays and reagents should help to elucidate the roles of TVLPs and toroviruses in diarrheal disease in humans.


title: The direction of the postural response to a vestibular perturbation is mediated by the cerebellar vermis.

text: When an electrical stimulus is applied to perturb the vestibular system, a postural response is generated orthogonal to head orientation. It has previously been shown that there is a convergence of neck proprioceptive and vestibular input within the cerebellum to provide a head-on-body reference frame (Manzoni et al. in Neuroscience 93:1095-1107, 1999). The objective of this experiment was to determine whether the direction of the postural response to a vestibular perturbation is modulated when function of the cerebellar vermis is temporarily depressed. Twenty participants were randomly assigned to a SHAM group (paired-pulse transcranial magnetic stimulation) or a TEST group (continuous theta burst stimulation). Stochastic vestibular stimulation (SVS) was applied to standing subjects with their head facing forward or over their left shoulder. Cumulant density traces were established between the SVS and shear force over 180°, and the peak amplitude determined the direction of sway. There were no significant changes in sway direction when the head was facing forward for either stimulation (TEST or SHAM; p = 0.889) or when the head was facing over the shoulder for the SHAM condition (p = 0.954). There was, however, a significant change in sway direction when the head was turned with a depressed cerebellum (p = 0.018); from the expected antero-posterior direction, orthogonal to head orientation, to one slightly more mediolateral with respect to the feet. These results suggest the cerebellum plays a role in the integration of input to generate an appropriately directed postural response relative to the head position.


title: Toxicology and Carcinogenesis Studies of Bromoethane (Ethyl Bromide) (CAS No. 74-96-4) in F344/N Rats and B6C3F1 Mice (Inhalation Studies).

text: Bromoethane is an alkylating agent used primarily as a chemical intermediate in various organic syntheses. In toxicology and carcinogenesis studies, groups of F344/N rats and B6C3F1 mice of each sex received whole-body exposure to bromoethane (greater than 98% pure) once for 4 hours or for 6 hours per day, 5 days per week, for 14 days, 14 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and Chinese hamster ovary (CHO) cells. Single-Exposure, Fourteen-Day, and Fourteen-Week Studies: Single-exposure inhalation studies were conducted in rats and mice at target concentrations of 625, 1,250, 2,500, 5,000, or 10,000 ppm bromoethane. All rats exposed to 10,000 ppm bromoethane and 3/5 female rats exposed to 5,000 ppm died before the end of the single-exposure studies. All mice exposed to 5,000 or 10,000 ppm bromoethane and 2/5 female mice exposed to 1,250 ppm died before the end of the studies. Fourteen-day inhalation studies were conducted in rats and mice at target concentrations of 0, 250, 500, 1,000, 2,000, or 4,000 ppm bromoethane. All rats and mice exposed to 2,000 or 4,000 ppm died before the end of the 14-day studies. Final mean body weights of exposed and control rats were similar. Fourteen-week inhalation studies were conducted in rats and mice at target concentrations of 0, 100, 200, 400, 800, or 1,600 ppm bromoethane. Four of 10 male and 2/10 female rats exposed to 1,600 ppm died before the end of the 14-week studies. The final mean body weights of rats exposed to 1,600 ppm were lower than the initial mean body weights. Compound-related lesions observed in rats at 1,600 ppm, but not at lower concentrations, included minimal atrophy of the thigh muscle, minimal-to-moderate multifocal mineralization in the cerebellum of the brain, minimal-to-severe hemosiderosis of the spleen, marked atrophy of the testis, and minimal-to- mild atrophy of the uterus. The effects in the testis and uterus are probably due to chemical-related loss in body weight during the studies. In mice, compound-related deaths included 3/10 male and 1/10 female mice exposed to 1,600 ppm, 1/9 males exposed to 800 ppm, and 1/10 males exposed to 400 ppm. The final mean body weights of male and female mice exposed to 1,600 ppm were about 15% lower than those of controls. Compound-related effects included atrophy of the uterus and involution of the ovary in females exposed to 1,600 ppm bromoethane. Based on these results, 2-year studies were conducted by exposing groups of 49 or 50 rats or mice of each sex to bromoethane at 0, 100, 200, or 400 ppm, 6 hours per day, 5 days per week. Body Weight and Survival in the Two-Year Studies: Mean body weights of exposed and control rats were generally similar throughout the studies. No significant differences in survival were observed between any groups of male rats (control, 17/49; 100 ppm, 26/50; 200 ppm, 27/50; 400 ppm, 21/50); survival of the 100-ppm group of female rats was greater than that of controls (19/50; 29/50; 24/49; 23/50), and the number of control and 400-ppm male rats and control female rats surviving to the end of the studies was low. Mean body weights of the 400-ppm group of male mice were up to 9% lower than those of controls throughout the study. Mean body weights of the 400-ppm group of female mice were generally 6%-16% lower than those of controls after week 29. No differences in survival were observed between any group of male mice (35/50; 37/50; 30/50; 34/50). The survival of the 400-ppm group of female mice was lower than that of controls at the end of the study (36/50; 37/50; 37/49; 23/49). Nonneoplastic and Neoplastic Effects in the Two-Year Studies: The incidences of pheochromocytomas or malignant pheochromocytomas (combined) of the adrenal medulla were increased in exposed male rats (control, 8/40; 100 ppm, 23/45; 200 ppm, 18/46; 400 ppm, 21/46). Granular cell neoplasms of the brain were seen in exposed male rats but not in controls (0/49; 3/50; 1/50; 1/50). A glioma, an astrocytoma, or an oligodendroglioma was seen in 3/50 male rats exposed to 100 p in 3/50 male rats exposed to 100 ppm. Gliomas were not observed in control female rats, but they occurred in exposed female rats with a significant positive trend (0/50; 1/50; 1/48; 3/50). The historical incidence of granular cell tumors in male F344/N rat chamber controls at the study laboratory is 0/297. The incidences of gliomas in the exposed female groups were not significantly greater than that in the controls and were within the historical incidence range for glial cell neoplasms for untreated controls in NTP studies (mean: 23/1,969, 1%; range: 0/50-3/50), but they exceeded the historical incidence range for chamber controls at the study laboratory (mean: 1/297, 0.3%; range: 0/50-1/50). Alveolar epithelial hyperplasia was increased in rats exposed to 400 ppm bromoethane (male: 3/48; 7/49; 7/48; 18/48; female: 5/50; 4/48; 5/47; 10/49). Alveolar/bronchiolar adenomas or carcinomas (combined) were seen in four male rats exposed to 200 ppm and in one exposed to 400 ppm. Alveolar/bronchiolar adenomas were observed in 3/49 female rats at 400 ppm but not at lower concentrations or in controls. The incidences in exposed male and female rats were not significantly greater than those in controls; however, the historical incidence in rat chamber controls for alveolar/bronchiolar adenomas or carcinomas (combined) at the study laboratory is 6/299 (2%) for males and 4/297 (1.3%) for females. The incidences of epithelial hyperplasia and squamous metaplasia of the nasal cavity were increased in rats exposed to 400 ppm. The incidence of suppurative inflammation of the nasal cavity was increased in exposed male rats, and the incidences of suppurative inflammation of the larynx and metaplasia of the olfactory sensory epithelium were increased in exposed male and female rats. An adenoma of the nose was seen in one 400-ppm male rat and in one 400-ppm female mouse. Suppurative inflammation and dilatation of the salivary gland ducts were observed at increased incidences in the 200- and 400-ppm groups of female rats. Animals were found to be positive for rat coronavirus/sialodacryoadenitis virus antibodies. The incidence of mammary gland neoplasms was significantly lower in female rats at 400 ppm than in controls (18/50; 15/50; 10/48; 7/50). Adenomas (0/50; 1/50; 1/47; 6/48), adenocarcinomas (0/50; 2/50; 3/47; 19/48), and squamous cell carcinomas (0/50; 1/50; 1/47; 3/48) of the uterus occurred in exposed female mice and not in control mice. The incidence of alveolar/bronchiolar neoplasms was greater in male mice at 400 ppm than in controls (adenomas or carcinomas, combined: 7/50; 6/50; 12/50; 15/50). Acute/chronic inflammation of the lung was observed at increased incidences in female mice at 200 and 400 ppm. Genetic Toxicology: Bromoethane, tested in a closed environment of a desiccator, was mutagenic in S. typhimurium strain TA100 with and without exogenous metabolic activation; it was not mutagenic in strain TA98. In cultured CHO cells, bromoethane induced sister chromatid exchanges (SCEs) but not chromosomal aberrations in both the presence and absence of exogenous metabolic activation. Conclusions: Under the conditions of these 2-year inhalation studies, there was some evidence of carcinogenic activity of bromoethane for male F344/N rats, as indicated by increased incidences of pheochromocytomas of the adrenal gland; neoplasms of the brain and lung may also have been related to exposure to bromoethane. For female F344/N rats, there was equivocal evidence of carcinogenic activity, as indicated by marginally increased incidences of neoplasms of the brain and lung. For male B6C3F1 mice, there was equivocal evidence of carcinogenic activity, based on marginally increased incidences of neoplasms of the lung. There was clear evidence of carcinogenic activity for female B6C3F1 mice, as indicated by neoplasms of the uterus. Synonyms: monobromoethane; bromic ether; hydrobromic ether


title: Identification and characterization of the woodchuck hepatitis virus origin of DNA replication.

text: Replication of the woodchuck hepatitis virus (WHV) genome requires the synthesis of minus-strand DNA from an RNA template, the pregenome, by reverse transcription. During this reaction, the 5' end of minus-strand DNA becomes covalently linked to a protein. The position of the 5' end of minus-strand DNA was identified previously, but the initiation site for DNA synthesis on pregenomic RNA remained ambiguous because of a sequence repetition at the termini of the RNA template for reverse transcription. Employing a recently designed expression vector for the production of infectious WHV, we localized the origin of minus-strand DNA synthesis to the 3' end of pregenomic RNA. In addition, we identified the nucleotide sequences on pregenomic RNA that provide the signal for the initiation of reverse transcription. Removal of this signal sequence from pregenomic RNA abolished minus-strand DNA synthesis. Insertion of a DNA oligomer bearing this signal sequence at the 3' end of pregenomic RNA restored the production of minus-strand DNA joined to protein. Our results support a model in which protein is the primer for reverse transcription of minus-strand DNA of WHV.


title: Analysis of efficiently packaged defective interfering RNAs of murine coronavirus: localization of a possible RNA-packaging signal.

text: We have previously shown that most of the defective interfering (DI) RNA of mouse hepatitis virus (MHV) are not packaged into virions. We have now identified, after 21 serial undiluted passages of MHV, a small DI RNA, DIssF, which is efficiently packaged into virions. The DIssF RNA replicated at a high efficiency on its transfection into the helper virus-infected cells. The virus released from the transfected cells interfered strongly with mRNA synthesis and growth of helper virus. cDNA cloning and sequence analysis of DIssF RNA revealed that it is 3.6 kb and consists of sequences derived from five discontinuous regions of the genome of the nondefective virus. The first four regions (domains I to IV) from the 5' end are derived from gene 1, which presumably encodes the RNA polymerase of the nondefective virus. The entire domain I (859 nucleotides) and the first 750 nucleotides of domain II are also present in a previously characterized DI RNA, DIssE, which is not efficiently packaged into virions. Furthermore, the junction between these two domains is identical between the two DI RNAs. The remaining 77 nucleotides at the 3' end of domain II and all of domains III (655 nucleotides) and IV (770 nucleotides) are not present in DIssE RNA. These four domains are derived from gene 1. In contrast, the 3'-most domain (domain V, 447 nucleotides) is derived from the 3' end of the genomic RNA and is also present in DIssE. The comparison of primary sequences and packaging properties between DIsse and DIssF RNAs suggested that domains III and IV and part of the 3' end of domain II contain the packaging signal for MHV RNA. This conclusion was confirmed by inserting these DIssF-unique sequences into a DIssE cDNA construct; the in vitro-transcribed RNA from this hybrid construct was efficiently packaged into virion particles. DIssF RNA also contains an open reading frame, which begins from domain I and ends at the 5'-end 20 bases of domain III. In vitro translation of DIssF RNA and metabolic labeling of the virus-infected cells showed that this open reading frame is indeed translated into a 75-kDa protein. The structures of both DIssE and DIssF RNAs suggest that a protein-encoding capability is a common characteristic of MHV DI RNA.


title: Detection of coronavirus 229E antibody by indirect hemagglutination.

text: Tannic-acid treated sheep erythrocytes (fresh or glutaraldehyde preserved) were sensitized with 229E antigens from human embryonic lung (RU-1) cell cultures. Indirect hemagglutination (IHA) antigen titers in 229E-infected cell cultures paralleled virus infectivity and complement fixation (CF) antigen titers. The identity of the IHA antigen was confirmed by testing extracts from inoculated and control cell cultures for ability to inhibit IHA. Also, significant increases in IHA antibody were demonstrated with acute and convalescent serum pairs from patients with proven 229E infections. A comparison of IHA, neutralization and CF titers for 229E antibodies was made on human sera drawn from different populations. The IHA and neutralization results were in agreement on 93% of the 129 sera found to be positive by at least one of three tests. The number of antibody titers detected by the CF test was insufficient to permit comparison. Hyperimmune sera from animals immunized with OC 43 did not react with 229E by IHA. Also no increase in IHA antibody was demonstrated with acute and convalescent serum pairs from patients with seroconversions to OC 43. These findings suggest that the IHA test provides (i) a rapid and sensitive method for serodiagnosis of 229E infections and (ii) a simple and inexpensive method for seroepidemiological studies.


title: Flavivirus membrane fusion.

text: Flavivirus membrane fusion is mediated by a class II viral fusion protein, the major envelope protein E, and the fusion process is extremely fast and efficient. Understanding of the underlying mechanisms has been advanced significantly by the determination of E protein structures in their pre- and post-fusion conformations and by the elucidation of the quarternary organization of E proteins in the viral envelope. In this review, these structural data are discussed in the context of functional and biochemical analyses of the flavivirus fusion mechanism and its characteristics are compared with those of other class II- and class I-driven fusion processes.


title: Development of PCR-based techniques to identify porcine transmissible gastroenteritis coronavirus isolates.

text: Sixteen isolates of transmissible gastroenteritis virus and one isolate of porcine respiratory coronavirus were characterized using RT-PCR amplification of 4 antigenic subsites in the site A epitope on the TGEV spike gene. The PCR products were digested with restriction enzymes Sau3AI and SspI and the sizes of the fragments were determined. Three different digestion patterns were observed with each enzyme. The recognition site for Sau3AI was missing in 1 isolate, was present in 13 isolates and 3 isolates had 2 sites. PCR-products with a single site had 3 different fragment sizes and the other isolates produced 2 fragments with different sizes. The SspI recognition site was not present in 5 isolates and 12 isolates had a single site that produced 2 fragments of different sizes. Based on the restriction fragment sizes, the 17 isolates were separated into 7 groups. Direct sequencing of the 455 bp nested set fragments demonstrated greater than 96% sequence homology among the 16 isolates and 100% homology in the 4 antigenic subsites in the conserved site A epitope. The groups are discussed in relation to their sequence homology and virulence. In vitro procedures have been developed to identify several porcine enteric coronavirus isolates at the strain level.


title: Heparin binding directs activation of T cells against adeno-associated virus serotype 2 capsid.

text: Activation of T cells to the capsid of adeno-associated virus (AAV) serotype 2 vectors has been implicated in liver toxicity in a recent human gene therapy trial of hemophilia B. To further investigate this kind of toxicity, we evaluated T-cell responses to AAV capsids after intramuscular injection of vectors into mice and nonhuman primates. High levels of T cells specific to capsids of vectors based on AAV2 and a phylogenetically related AAV variant were detected. Vectors from other AAV clades such as AAV8 (ref. 3), however, did not lead to activation of capsid-specific T cells. Through the generation of AAV2-AAV8 hybrids and the creation of site-directed mutations, we mapped the domain that directs the activation of T cells to the RXXR motif on VP3, which was previously shown to confer binding of the virion to heparan sulfate proteoglycan (HSPG). Evaluation of natural and engineered AAV variants showed direct correlations between heparin binding, uptake into human dendritic cells (DCs) and activation of capsid-specific T cells. The role of heparin binding in the activation of CD8(+) T cells may be useful in modulating the immunogenicity of antigens and improving the safety profile of existing AAV vectors for gene therapy.


title: Respiratory syncytial and other virus infections in persons with chronic cardiopulmonary disease.

text: Respiratory syncytial virus (RSV) has been increasingly recognized as an important cause of serious respiratory illness in some adult populations, including those with underlying cardiopulmonary diseases. However, the precise incidence and the clinical impact of RSV in this group are unknown. Therefore, the incidence and clinical impact of RSV infection in persons with chronic obstructive pulmonary disease (COPD) and congestive heart failure (CHF) who reside in the community were prospectively evaluated over two consecutive winters in 134 persons. Eight RSV (incidence of 4.3 per 100 subject-winters), 13 influenza A (incidence of 7.0 per 100 subject-winters), seven rhinovirus, nine coronavirus, and two parainfluenza virus infections were identified. The clinical illnesses associated with RSV and influenza A virus were similar, causing both upper and lower respiratory signs and symptoms. The clinical impact was significant as three of eight RSV-infected subjects were hospitalized compared with six of 13 influenza A-infected persons and zero of seven rhinovirus-infected persons.


title: Elimination of transmissible gastroenteritis virus from a pig farm by culling and serological surveillance.

text: The first and only outbreak of transmissible gastroenteritis in pigs in Ireland occurred in 1984 in a 650-sow unit which was one of three such units owned by one enterprise which supplied pigs to a 12,000 head fattening unit in the same ownership. The depopulation of the clinically infected unit and extensive serological monitoring of pigs in the other units and in pig farms throughout the country has proved successful in the elimination of transmissible gastroenteritis infections from pigs in Ireland.


Emerging infections, transfusion safety, and epidemiology.


title: Large-scale spatial-transmission models of infectious disease.

text: During transmission of seasonal endemic diseases such as measles and influenza, spatial waves of infection have been observed between large distant populations. Also, during the initial stages of an outbreak of a new or reemerging pathogen, disease incidence tends to occur in spatial clusters, which makes containment possible if you can predict the subsequent spread of disease. Spatial models are being used with increasing frequency to help characterize these large-scale patterns and to evaluate the impact of interventions. Here, I review several recent studies on four diseases that show the benefits of different methodologies: measles (patch models), foot-and-mouth disease (distance-transmission models), pandemic influenza (multigroup models), and smallpox (network models). This review highlights the importance of the household in spatial studies of human diseases, such as smallpox and influenza. It also demonstrates the need to develop a simple model of household demographics, so that these large-scale models can be extended to the investigation of long-time scale human pathogens, such as tuberculosis and HIV.


title: Severe Thrombocytopenia in a Patient with COVID-19.

text: Coronavirus disease 2019 (COVID-19) outbreak is spreading rapidly all over the world, being a major threat to public health. Since clinical feature of COVID-19 has not been fully evaluated yet, empirical antibacterial agents are frequently combined for the treatment of COVID-19 in addition to antiviral agents, concerning co-existing bacterial pathogens. We experienced a case of severe thrombocytopenia with epistaxis and petechiae, while treating a COVID-19 patient with ceftriaxone, levofloxacin, and lopinavir/ritonavir. The platelet count decreased to 2,000/mm³ and recovered after discontinuation of the three suspected drugs. In treating a potentially fatal emerging infectious disease, empirical and/or experimental approach would be unavoidable. However, the present case suggests that the possibility of adverse effects caused by polypharmacy should also be carefully considered.


title: [Infectious diseases].

text: The recommendations for the treatment of gonorrhea have been changed: ceftriaxone 500 mg IM plus azithromycin 1 g PO is recommended. Prophylaxis of recurrent cellulitis with penicillin 250 mg 2 x/d PO may be considered. E. coli ESBL does not require contact isolation anymore. Fecal transplantation seems so far to be the most effective treatment of recurrent C. dificile. Two new respiratory viruses, Middle East Coronavirus (MERS-CoV) and avian-origin Influenza A (H7N9) have been reported. Oral valganciclovir treatment reduces the risk of hearing loss in congenital CMV infection. An outbreak of mould infections of the central nervous system has been described in the United States following injection of contaminated steroids.


title: Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus.

text: A previously undescribed coronavirus (CoV) is the etiologic agent responsible for severe acute respiratory syndrome (SARS). Using a panel of contiguous cDNAs that span the entire genome, we have assembled a full-length cDNA of the SARS-CoV Urbani strain, and have rescued molecularly cloned SARS viruses (infectious clone SARS-CoV) that contained the expected marker mutations inserted into the component clones. Recombinant viruses replicated as efficiently as WT virus and both were inhibited by treatment with the cysteine proteinase inhibitor (2S,3S)-transepoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester. In addition, subgenomic transcripts were initiated from the consensus sequence ACGAAC in both the WT and infectious clone SARS-CoV. Availability of a SARS-CoV full-length cDNA provides a template for manipulation of the viral genome, allowing for the rapid and rational development and testing of candidate vaccines and therapeutics against this important human pathogen.


title: [A follow-up study of the lung function and the chest CT changes in medical staff with severe acute respiratory syndrome in Beijing].

text: OBJECTIVE To analyze the lung function and radiological changes in rehabilitating severe acute respiratory syndrome (SARS) patients of medical staff in Beijing. METHODS Follow-up lung function tests and chest high-resolution computerized tomography (HRCT) were performed in medical staff with SARS from Dec. 2003 to Feb. 2004. RESULTS Thirty-one (7.64%) of 406 patients showed abnormal ventilatory function, while 165 of 404 patients showed diffusing abnormality. Of the 434 patients who had received HRCT scans, 151 (34.79%) showed abnormalities including subpleural and basal ground-glass and reticular attenuation, nodular septal thickening and bronchiectasis. Of the 395 patients who had received both lung function test and HRCT scanning, 55 (13.92%) had abnormalities both in the lung function and HRCT. The decrease in diffusing capacity in patients with HRCT changes was more significant than those without HRCT changes. CONCLUSIONS Lung function and lung imaging were abnormal in some patients with SARS after recovery. It is suggested that the lung damage is chronic, and follow-up is needed.


title: Clinical outcomes of single incision laparoscopic cholecystectomy in the anglophone Caribbean: a multi centre audit of regional hospitals.

text: INTRODUCTION There has been no report on Single-Incision Laparoscopic Surgery (SILS) cholecystectomy outcomes since it was first performed in the Anglophone Caribbean in 2009. METHODS A retrospective audit evaluated the clinical outcomes of SILS cholecystectomies at regional hospitals in the 17 Anglophone Caribbean countries. Any cholecystectomy using a laparoscopic approach in which all instruments were passed through one access incision was considered a SILS cholecystectomy. The following data were collected: patient demographics, indications for operation, intraoperative details, surgeon details, surgical techniques, specialized equipment, conversions, morbidity and mortality. Descriptive statistics were generated using SPSS 12.0. RESULTS There were 85 SILS cholecystectomies in women at a mean age of 37.4 ± 8.5 years with a mean BMI of 30.9 ± 2.8. There were 59 elective and 26 emergent cases. Specialized access platforms were used in the first 35 cases and reusable instruments were passed directly across fascia in the latter 50 cases. The mean operative time was 62.9 ± 17.9 minutes. There was no mortality, 2 conversions to multi-trocar laparoscopy and 5 minor complications. Ambulatory procedures were performed in 43/71 (60.6%) patients scheduled for elective operations. CONCLUSION In the Caribbean setting, SILS cholecystectomy is a feasible and safe alternative to conventional multi-trocar laparoscopic cholecystectomy for gallbladder disease.


title: Computer-aided diagnosis of renal obstruction: utility of log-linear modeling versus standard ROC and kappa analysis.

text: BACKGROUND: The accuracy of computer-aided diagnosis (CAD) software is best evaluated by comparison to a gold standard which represents the true status of disease. In many settings, however, knowledge of the true status of disease is not possible and accuracy is evaluated against the interpretations of an expert panel. Common statistical approaches to evaluate accuracy include receiver operating characteristic (ROC) and kappa analysis but both of these methods have significant limitations and cannot answer the question of equivalence: Is the CAD performance equivalent to that of an expert? The goal of this study is to show the strength of log-linear analysis over standard ROC and kappa statistics in evaluating the accuracy of computer-aided diagnosis of renal obstruction compared to the diagnosis provided by expert readers. METHODS: Log-linear modeling was utilized to analyze a previously published database that used ROC and kappa statistics to compare diuresis renography scan interpretations (non-obstructed, equivocal, or obstructed) generated by a renal expert system (RENEX) in 185 kidneys (95 patients) with the independent and consensus scan interpretations of three experts who were blinded to clinical information and prospectively and independently graded each kidney as obstructed, equivocal, or non-obstructed. RESULTS: Log-linear modeling showed that RENEX and the expert consensus had beyond-chance agreement in both non-obstructed and obstructed readings (both p < 0.0001). Moreover, pairwise agreement between experts and pairwise agreement between each expert and RENEX were not significantly different (p = 0.41, 0.95, 0.81 for the non-obstructed, equivocal, and obstructed categories, respectively). Similarly, the three-way agreement of the three experts and three-way agreement of two experts and RENEX was not significantly different for non-obstructed (p = 0.79) and obstructed (p = 0.49) categories. CONCLUSION: Log-linear modeling showed that RENEX was equivalent to any expert in rating kidneys, particularly in the obstructed and non-obstructed categories. This conclusion, which could not be derived from the original ROC and kappa analysis, emphasizes and illustrates the role and importance of log-linear modeling in the absence of a gold standard. The log-linear analysis also provides additional evidence that RENEX has the potential to assist in the interpretation of diuresis renography studies.


title: A prospective study of preoperative computed tomographic angiographic mapping of free fibula osteocutaneous flaps for head and neck reconstruction.

text: BACKGROUND In designing an osteocutaneous fibula flap, poor planning, aberrant anatomy, or inadequate perforators may necessitate modification of the flap design, exploration of the contralateral leg, or additional flap harvest. The authors studied the predictive power of computed tomographic angiography in osteocutaneous fibula flap planning and execution. METHODS The authors studied a prospective cohort of 40 consecutive patients who underwent preoperative computed tomographic angiography mapping of the peroneal artery and its perforators and subsequent free fibula flap reconstruction of mandibular or maxillary defects. The authors compared their analysis of perforator anatomy, peroneal artery origin, and fibula length with intraoperative clinical findings. RESULTS Overall, computed tomographic angiography identified 94.9 percent of the cutaneous perforators found intraoperatively. Clinically, perforators were located an average of 8.7 mm from their predicted locations. The peroneal artery origin from the tibioperoneal trunk averaged 6.0 mm from its predicted location. The average length of the fibula differed from the predicted length by 8.0 mm. Computed tomographic angiography accurately predicted perforators as either septocutaneous or musculocutaneous 93.0 percent of the time. Perforator size was accurately predicted 66.7 percent of the time. Skin islands and osteotomies were modified in 25.0 percent of the cases on the basis of computed tomographic angiography findings. Two patients had hypoplastic posterior tibial arteries, prompting selection of the contralateral leg. There were no total flap or skin paddle losses. CONCLUSIONS Computed tomographic angiography accurately predicted the course and location of the peroneal artery and perforators; perforator size was less accurately estimated. Computed tomographic angiography provides valuable information to facilitate osteocutaneous fibula flap harvest.


title: Asthma prevalence and control characteristics by race/ethnicity--United States, 2002.

text: During 1980-1999, asthma prevalence, morbidity, and mortality increased among U.S. adults. These annual rates were higher among certain racial/ethnic minority populations than among whites. In addition, racial/ethnic minority populations reported higher use of emergency departments (EDs) and doctors' offices for asthma treatment than whites. To assess asthma prevalence and asthma-control characteristics among racial/ethnic populations, CDC analyzed 2002 data from the Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the results of that analysis, which indicated that among the estimated 16 million (7.5%) U.S. adults with asthma, self-reported current asthma prevalence among racial/ethnic minority populations ranged from 3.1% to 14.5%, compared with 7.6% among whites. Comprehensive state-specific asthma surveillance data are necessary to identify disparities in asthma prevalence and asthma-control characteristics among racial/ethnic populations and to develop targeted public health interventions.


title: Axonal damage is T cell mediated and occurs concomitantly with demyelination in mice infected with a neurotropic coronavirus.

text: Mice infected with mouse hepatitis virus (MHV) strain JHM develop primary demyelination. Herein we show that axonal damage occurred in areas of demyelination and also in adjacent areas devoid of myelin damage. Immunodeficient MHV-infected RAG1-/- mice (mice defective in recombinase activating gene 1 expression) do not develop demyelination unless they receive splenocytes from a mouse previously immunized against MHV (G. F. Wu, A. Dandekar, L. Pewe, and S. Perlman, J. Immunol. 165:2278-2286, 2000). In the present study, we show that adoptive transfer of T cells was also required for the majority of the axonal injury observed in these animals. Both demyelination and axonal damage were apparent by 7 days posttransfer. Recent data suggest that axonal injury is a major factor in the long-term disability observed in patients with multiple sclerosis. Our data demonstrate that immune system-mediated damage to axons is also a common feature in mice with MHV-induced demyelination. Remarkably, there appeared to be a minimal, if any, interval of time between the appearance of demyelination and that of axonal injury.


title: The 3a Protein of SARS-coronavirus Induces Apoptosis in Vero E6 Cells.

text: An outbreak of severe acute respiratory syndrome (SARS) occurred in China and the first case emerged in mid November 2002. The etiologic agent of this disease was found to be a previously unknown coronavirus, SARS-CoV. The detailed pathology of SARS-CoV infection and the host response to the viral infection are still not known. The 3a gene encodes a non-structural viral protein which is predicted to be a transmembrane protein. In this study, we showed that the 3a protein was localized to the endoplasmic reticulum (ER) in 3a-transfected monkey kidney Vero E6 cells. In vitro experiments of chromatin condensation and DNA fragmentation suggest that the 3a protein may trigger apoptosis. Our data show that over-expression of a single SARS-CoV protein can induce apoptosis in vitro. Thus GFP-3a fusion protein could also be used as a biosensor for monitoring the cytopathic features of SARS infection, e.g. lymphopenia, in animal model systems, similar to nucleocapsid and 7a proteins.


title: Heptad repeat-derived peptides block protease-mediated direct entry from the cell surface of severe acute respiratory syndrome coronavirus but not entry via the endosomal pathway.

text: The peptides derived from the heptad repeat (HRP) of severe acute respiratory syndrome coronavirus (SCoV) spike protein (sHRPs) are known to inhibit SCoV infection, yet their efficacies are fairly low. Recently our research showed that some proteases facilitated SCoV's direct entry from the cell surface, resulting in a more efficient infection than the previously known infection via endosomal entry. To compare the inhibitory effect of the sHRP in each pathway, we selected two sHRPs, which showed a strong inhibitory effect on the interaction of two heptad repeats in a rapid and virus-free in vitro assay system. We found that they efficiently inhibited SCoV infection of the protease-mediated cell surface pathway but had little effect on the endosomal pathway. This finding suggests that sHRPs may effectively prevent infection in the lungs, where SCoV infection could be enhanced by proteases produced in this organ. This is the first observation that HRP exhibits different effects on virus that takes the endosomal pathway and virus that enters directly from the cell surface.


Cytological and microbiological results from equine guttural pouch lavages obtained percutaneously: correlation with histopathological findings.


docid: wxvmwkis

title: Molecular modeling studies of peptide drug candidates against SARS.

text: Flexible alignment and docking studies were conducted for the three octapeptides, ATLQANEV, AVLQSGFR, and ATLQAIAS, that were cleavable by SARS-CoV Mpro. It has been observed that all pharmacophores of the three peptides overlap very well, and that ATLQANEV binds best with the receptor, followed by AVLQSGFR, and then ATLQAIAS. During the process of docking the octapeptides to the SARS enzyme, the residues of the catalytic dyad, i.e., His-41 and Cys-145 are actively involved in forming the hydrogen bonds, so is the center residue (Gln) of all the three octapeptides. The findings are fully consistent with experimental observations. The present studies suggest that the octapeptides ATLQANEV and ATLQAIAS, like AVLQSGFR, might also be the good starting points for designing potential drugs against SARS.


title: Rhinoviruses as pathogens of the lower respiratory tract.

text: Rhinoviruses (RVs) are the most common upper respiratory pathogens, inducing the majority of common colds worldwide. RV-related morbidity, although significant cumulatively, has been considered trivial for the individual patient. However, recent strong epidemiological associations of RVs with asthma exacerbations, including severe episodes requiring hospitalization, indicate that RV infections can result in serious disease. Current evidence supports the possibility that RVs infect the lower airways, inducing a local inflammatory response. Such evidence suggests that the role of RVs in other lower respiratory diseases, such as pneumonia, bronchitis, bronchiolitis and cystic fibrosis, should be re-examined with polymerase chain reaction-based methodologies, which are considerably more sensitive than traditional, cell culture-based techniques. The mechanisms through which RVs induce lower airway disease are studied to understand the relative contributions of the epithelial, neurogenic and immune components in the antiviral response, and to permit the design and implementation of specific treatments.