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.


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.


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.


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.


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.


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.


title: Quantifying inadvertent thermal bowel injury from the monopolar instrument.

text: BACKGROUND Insulation defects are observed in 3-39 % of laparoscopic instruments. Electrosurgical injuries due to insulation defects or capacitive coupling remain an issue in laparoscopic surgery with a prevalence of 0.6-5 per thousand cases. Shielded instruments with active electrode monitoring (AEM) have been postulated to prevent these injuries. The benefit of these instruments has not been quantified. Most bowel injuries are unrecognized intra-operatively. Injury is revealed only after the patient exhibits peritonitis symptoms and surgical intervention to repair the bowel is required. These injuries may result in devastating and costly complications or mortality. The extent of bowel injury possible with commonly used generator settings and associated energy output has never been histologically defined. Our objectives in this experimental study were: quantify and compare the energy released through insulation defects or capacitive coupling with standard unshielded monopolar versus shielded instruments with (AEM), determine energy required to cause a visible burn, and relate the histological burn depth to a given amount of energy. METHODS Ex vivo porcine jejunum was used for tissue testing. An oscilloscope measured energy output from three common electrosurgical generators at recommended power settings with standard or AEM instruments with insulation defects and in capacitive coupling scenarios. Presence of a visible burn was noted, and depth of tissue damage for a given amount of energy was measured histologically. RESULTS All samples that received ≥3.8 J of energy had visible burns. As little as 10 J caused full wall thickness burns. 3.8 J was exceeded at the 30- and 50-W power settings in every experimental scenario using standard monopolar instruments; AEM instruments never approached this much energy. CONCLUSIONS Serious burn injury results from small amounts of energy leaked from standard instruments. AEM instruments appeared protective and did not leak sufficient energy to cause burn injuries to the bowel.


Quality improvement guidelines for the treatment of lower extremity deep vein thrombosis with use of endovascular thrombus removal.


title: Tunisian Recommendations for resumption of Reproductive Medicine activity in the Covid-19 pandemic.

text: The activity of the Reproductive Medicine poses a dilemma in this pandemic Covid-19. In fact, this is a theoretically non-emergency activity except for fertility preservation with oncological reasons. The majority of fertility societies in the world such as the American Society for Reproductive Medicine (ASRM) and the European Society of Human Reproduction and Embryology (ESHRE) recommended stopping the inclusion of new patients and continuing only the In Vitro Fertilization (IVF) cycles that have already been initiated by promoting Freeze-all as much as possible. Initilaly, the "Société Tunisienne de Gynécologie Obstétrique" (STGO) issued national recommendations that echo the international recommendations. These recommendations were followed by the majority of IVF center in Tunisia. However, a number of new data are prompting us to update these recommendations.


Public health then and now: celebrating 50 years of MMWR at CDC. Introduction.


title: Endoscopic management of high-grade dysplasia and intramucosal carcinoma: experience in a large academic medical center.

text: BACKGROUND Esophagectomy has been the standard treatment for Barrett's esophagus (BE) with high-grade dysplasia (HGD) and intramucosal cancer (IMC). Recently, endoscopic mucosal resection (EMR) and radiofrequency ablation (RFA) have become the preferred treatment for these patients in some centers. We report a single institution series of patients undergoing endoscopic management of HGD and IMC. METHODS Nineteen patients underwent endoscopic treatment for HGD or IMC between 2009 and 2012. The primary outcome measure was progression of BE necessitating esophagectomy. Secondary outcomes included complete eradication of intestinal metaplasia (CE-IM), complete eradication of dysplasia (CE-D), recurrence or progression of BE or dysplasia, and complications. Patients were followed for a median follow-up interval of 19 months following completion of RFA treatment. RESULTS Three patients (16 %) had a presenting diagnosis of IMC, and 16 (84 %) were treated for HGD. Twelve (63 %) had long-segment BE; the median length of BE was 5 cm. Ten (53 %) patients underwent EMR prior to RFA. CE-D was achieved in 88 % of patients, and CE-IM was achieved in 65 % of patients. A median of 2 (1-7) treatments were required, and there were no immediate post-procedure complications. Two patients developed recurrent dysplasia following complete eradication of BE, and each case was successfully managed with repeat RFA. Three patients (16 %) required esophagectomy within 6 months following RFA. A complete surgical resection was achieved in each case, and none of the patients developed lymph node metastases. CONCLUSIONS Complete eradication of HGD and IMC can be achieved via endoscopic therapy, thus avoiding esophagectomy in the majority of patients. However, a subset of patients will fail this treatment approach and will require surgical resection. With aggressive endoscopic treatment and surveillance, these patients can be identified at an early stage while curative resection is still possible.


title: Discovery of an RNA virus 3'->5' exoribonuclease that is critically involved in coronavirus RNA synthesis.

text: Replication of the giant RNA genome of severe acute respiratory syndrome (SARS) coronavirus (CoV) and synthesis of as many as eight subgenomic (sg) mRNAs are mediated by a viral replicase-transcriptase of outstanding complexity that includes an essential endoribonuclease activity. Here, we show that the CoV replicative machinery, unlike that of other RNA viruses, also uses an exoribonuclease (ExoN) activity, which is associated with nonstructural protein (nsp) 14. Bacterially expressed forms of SARS-CoV nsp14 were shown to act on both ssRNAs and dsRNAs in a 3'-->5' direction. The activity depended on residues that are conserved in the DEDD exonuclease superfamily. The protein did not hydrolyze DNA or ribose-2'-O-methylated RNA substrates and required divalent metal ions for activity. A range of 5'-labeled ssRNA substrates were processed to final products of approximately 8-12 nucleotides. When part of dsRNA or in the presence of nonlabeled dsRNA, the 5'-labeled RNA substrates were processed to significantly smaller products, indicating that binding to dsRNA in cis or trans modulates the exonucleolytic activity of nsp14. Characterization of human CoV 229E ExoN active-site mutants revealed severe defects in viral RNA synthesis, and no viable virus could be recovered. Besides strongly reduced genome replication, specific defects in sg RNA synthesis, such as aberrant sizes of specific sg RNAs and changes in the molar ratios between individual sg RNA species, were observed. Taken together, the study identifies an RNA virus ExoN activity that is involved in the synthesis of multiple RNAs from the exceptionally large genomic RNA templates of CoVs.


title: Proteolytic processing at the amino terminus of human coronavirus 229E gene 1-encoded polyproteins: identification of a papain-like proteinase and its substrate.

text: Expression of the coronavirus gene 1-encoded polyproteins, pp1a and pp1ab, is linked to a series of proteolytic events involving virus-encoded proteinases. In this study, we used transfection and immunoprecipitation assays to show that the human coronavirus 229E-encoded papain-like cysteine proteinase, PCP1, is responsible for the release of an amino-terminal protein, p9, from the gene 1-encoded polyproteins. The same protein, p9, has also been identified in virus-infected cells. Furthermore, using an in vitro trans-cleavage assay, we defined the proteolytic cleavage site at the carboxyl terminus of p9 as pp1a-pp1ab amino acids Gly-111 and Asn-112. These results and a comparative sequence analysis suggest that substrate positions P1 and P5 seem to be the major determinants of the PCP1 cleavage site and that the latter can occupy a variable position at the amino terminus of the coronavirus ppla and pplab polyproteins. By combining the trans-cleavage assay with deletion mutagenesis, we were also able to locate the boundaries of the active PCP1 domain between pp1a-pp1ab amino acids Gly-861-Glu-975 and Asn-1209-Gln-1285. Finally, codon mutagenesis was used to show that Cys-1054 and His-1205 are essential for PCP1 proteolytic activity, suggesting that these amino acids most likely have a catalytic function.


title: Front-line ownership: generating a cure mindset for patient safety.

text: Great advances have been made in standardization and human factors engineering that have reduced variability and increased reliability in healthcare. As important as these advances are, the authors believe there is another important but largely ignored layer to the safety story in healthcare that has prevented us from progressing. In the field of infection prevention and control (IPAC), despite great attempts over several decades to improve compliance with hand hygiene, surveillance, environmental cleaning, isolation protocols and other control measures, very significant challenges remain. We believe this failure is in part due to the power gradients, often dysfunctional relationships and lack of safety mindfulness that exist in hospitals and healthcare more generally. Furthermore, safety culture requires different approaches and considerable ongoing attentiveness. If this is the case, and the authors contend in this paper that it is, then the role of the front line is much more important than many of our healthcare safety and IPAC approaches suggest.


title: Long distance RT-PCRs of human coronavirus 229E RNA.

text: The generation and cloning of cDNA fragments longer than 10 kb is often a difficult and time consuming task. In this study, we have analysed the conditions necessary of produce reverse transcripts longer than 10 kb that can be amplified by polymerase chain reaction. Thus, we isolated poly(A)-RNA from human coronavirus 229E infected MRC-5 cells and did reverse transcription using a sequence-specific primer. Subsequently, we amplified PCR products of varying length upstream of the primer position. Optimisation of the poly(A)-RNA preparation, the reverse transcription protocol and the polymerase chain reaction cycle conditions enabled us to successfully amplify regions of the human coronavirus 229E genome between 11.5 and 20.3 kb in length.


title: Are US jurisdictions prepared to dispense medical countermeasures through open points of dispensing? Findings from a national study.

text: Communities will rely on open points of dispensing (PODs) for mass dispensing of medical countermeasures following a bioterrorism attack or a pandemic. US Cities Readiness Initiative (CRI) open POD preparedness is assessed using the Technical Assistance Review (TAR) but focuses on oral prophylaxis dispensing; mass vaccination readiness is not well measured. Non-CRI preparedness had not been studied. In 2013 an online questionnaire was sent to all 456 CRIs and a random sample of 500 non-CRIs to measure open POD preparedness and exercise participation. Hierarchical linear regression was used to describe factors associated with higher POD preparedness and exercise participation scores. In total, 257 subjects participated, for a 41% response rate. Almost all open PODs have existing written plans and/or a layout for each site (93.4%, n=240, and 87.0%, n=220). Only half (46.7%, n=120) have an alternative dispensing modality in place, and even fewer (42.6%, n=104) report having adequate staffing. Determinants of open POD preparedness were perceived preparedness, participation in more POD exercises, and more closed POD coverage. Most jurisdictions conducted a full-scale exercise and a staff notification drill (83.7%, n=215 for both). Fewer than half (40.5%, n=104) have conducted a vaccination clinic exercise. Determinants of increased POD exercises were perceived preparedness, years of work experience, community type (nontribal), and larger population. Because successful open POD deployment is critical, jurisdictions need to plan for mass vaccination, use of alternative dispensing modalities, and recruitment strategies to increase POD staffing.


Simultaneous detection of SARS coronavirus and influenza A viruses using real-time polymerase chain reaction.


title: Contribution of selected metabolic diseases to early childhood deaths--Virginia, 1996-2001.

text: Sudden infant death syndrome (SIDS), or the death of an infant aged <1 year that remains unexplained after a thorough investigation, is the third most common cause of death among infants in the United States. Sudden, unexplained deaths also occur among children aged >/=1 year; however, the number of these deaths is not well documented. Certain cases of SIDS and sudden unexplained death beyond infancy might be attributable to complications of unrecognized metabolic diseases. Tandem mass spectrometry (tandem MS) can be used to screen for several of these disorders. Despite the low prevalence of these diseases, newborn screening for these disorders has been found to compare favorably with the cost of other screening programs. However, the contribution of these diseases to early childhood deaths is not well understood. To determine the proportion of sudden, unexpected early childhood deaths associated with selected metabolic diseases, CDC, the Office of the Chief Medical Examiner (ME) in Virginia, and a private laboratory conducted a population- based study. This report summarizes the results of the study, which indicate that 1% of children had a positive postmortem metabolic screen using tandem MS. Of the eight children with positive screening tests, seven might have had improved outcomes had they been identified and treated during the newborn period. The use of tandem MS in newborn screening programs could offer an opportunity to prevent early childhood mortality.


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title: The role of infectious aerosols in disease transmission in pigs.

text: Airborne transmission is of significance for a number of infectious diseases in pigs. The general principles of the airborne pathway, including aerosol production, decay and inhalation, are reviewed. Practical issues regarding aerosol sampling and sample analysis are also discussed. Details of the aerobiology of porcine diseases, including foot-and-mouth disease, Aujesky's disease, and respiratory diseases, are explained. Some additional, potentially airborne diseases are discussed in terms of the evidence for their aerosol transmission. In order to prevent airborne diseases in pigs, dust reduction, air filtration, air disinfection, vaccination and the establishment of disease-free regions could be considered.


title: Turkey origin reovirus-induced immune dysfunction in specific pathogen free and commercial turkey poults.

text: Recently, pathogenesis studies, using genetically distinct turkey-origin reoviruses (TRVs), revealed that poults infected with certain TRV isolates had moderate to severe bursal atrophy, suggesting virus-induced immune dysfunction. In order to characterize the effect of TRV infection on the turkey immune system, classical assays were undertaken to quantify the humoral and cell-mediated immune responses in small Beltsville and broad-breasted white poults infected with the TRV isolate NC/SEP-R44/03. A marked effect on the cutaneous basophil hypersensitivity response, and on the antibody response to Newcastle disease virus (NDV) exposure, was noted in commercial and specific pathogen free (SPF) poults inoculated with NC/SEP-R44/03 at three days of age. Moderate to severe bursal atrophy, similar to that noted previously in SPF poults, occurred in commercial poults inoculated at three days of age. This immune dysfunction and bursal atrophy was not present in commercial poults inoculated at three weeks of age.


title: Propagation of bovine coronavirus on Vero cell line: electron microscopic studies.

text: The in vitro adaptation of enteric coronaviruses is difficult. In our Laboratories, the multiplication of the COI bovine strain (F15 origin), on Vero cells is commonly practised. Since other laboratory tests (hemagglutination inhibition, cytopathic effect and immunofluorescence) are not totally reliable, electron microscopic studies present definite advantages in the interpretation of viral synthesis, as demonstrated at the 11th passage in our study. We were able to observe the following: --that the rate of infected and productive cells was important; --that the stages of viral morphogenesis were well established; --that the viral synthesis was prolific and regular and could be witnessed by important intracytoplasmic storage. The results of this study prove that the choice of our cell system is very satisfactory.


title: Infectious bronchitis virus field vaccination coverage and persistence of Arkansas-type viruses in commercial broilers.

text: To determine the coverage of infectious bronchitis virus (IBV) vaccine field boost in commercial broilers, estimate the relative amount of vaccine virus in the trachea, and follow the clearance of the vaccine, we collected approximately 100 tracheal swabs at various times postvaccination from 10 different flocks and used real-time reverse transcriptase-PCR (RT-PCR) to detect the virus. This allowed us to detect vaccine virus in as few as 3% of the birds in a flock of 20,000 birds with a 95% confidence level. We found that the number of birds positive for IBV vaccine following vaccination in the field resembled a parabolic-shaped curve that peaked around 14 days postvaccination, or it resembled a sinusoidal-type wave with a frequency of about 2 wk. The patterns did not appear to correlate with water or spray vaccination methods, nor did they correlate with the type of backpack sprayer used. The highest number of positive birds in a flock ranged from 66% to 100%. The viral genome copies in the tracheal swabs, as determined by real-time RT-PCR, ranged from 1 x 10(2.6)/ml to 1 x 10(5.2)/ml and, in most studies, had a positive correlation with the number of birds positive for vaccine virus in the flock. On the last sample day of each study, 21, 28, or 35 days postvaccination, from 12% to 66% of the birds were still positive for vaccine virus, and although different IBV vaccine types were used in each study, only Arkansas vaccine virus was identified in selected samples on those days. Arkansas vaccine virus was also the only virus identified in selected samples at 1, 3, and 5 days postvaccination, clearly indicating that Arkansas vaccine virus is persisting in the birds. Protection studies conducted on birds vaccinated with Arkansas- and Delaware-type vaccines and removed from the field at 21 days postvaccination showed complete protection against challenge with Delaware (except for one bird), whereas protection against Arkansas challenge was between 37.5% and 62.5%. Our findings show that introduction of IBV vaccines into a commercial broiler flock do not necessarily follow a seemingly logical pattern of a high number of birds infected followed by clearance from the trachea, but resembled either a parabolic curve or a sinusoidal-type wave. In addition, Arkansas vaccine viruses are clearly persisting in commercial broilers, which may be because of incomplete protection observed for that IBV type.


title: Mutational analysis of fusion peptide-like regions in the mouse hepatitis virus strain A59 spike protein.

text: The coronavirus peplomer protein S is responsible for attachment and fusion during viral entry as well as for the induction of cell to cell fusion. While several regions within S have been shown to influence the ability to induce fusion, the region of the protein actually responsible for fusion, the fusion peptide, has not yet been identified. We identified two hydrophobic peptides (peptides #1 and #2) within MHV-A59 S2 as possible fusion domains. This was based on hydrophobicity, conservation among coronavirus S proteins and the prediction of a sided helix conformation. Using site directed mutagenesis and an in vitro cell to cell fusion assay we showed that substitution of hydrophobic amino acids with charged amino acids, within the predicted hydrophobic face of either of these two peptides eliminated fusion. Within peptide #1 substitution of the same hydrophobic amino acids with other hydrophobic amino acids or substitution of polar amino acids with charged or polar amino acids had little effect on fusion. Thus peptides #1 and #2 remain likely candidates for the MHV fusion peptide. A third previously identified peptide within S2 (Chambers et al., 1990) is unlikely as a fusion peptide as it is not well conserved among coronaviruses and substitution within the hydrophobic face with charged amino acids does not effect fusion.


title: Isolation and propagation of a human enteric coronavirus.

text: Coronavirus-like particles were found by electron microscopy in stools from infants with necrotizing enterocolitis. Stool samples from these infants as well as control specimens were passaged in cultures of human fetal intestinal organs. Two samples yielded virus-like particles and these have now been passaged 14 times (HEC 14). Gradient-purified HEC 14 strains had typical coronavirus morphology on electron microscopy and contained five major proteins with molecular sizes ranging from 190 to 23 kilodaltons. Infants with necrotizing enterocolitis developed specific antibody to the viral antigens between the acute and convalescent stages of the disease, as shown by examining serum specimens by single radial hemolysis, immunoenzymatic assay, and Western immunoblotting. No cross-reactivity was shown with other coronavirus strains tested, or with the newly isolated viruses of the Breda-Berne group, responsible for calf or horse diarrhea.


title: [Etiological diagnosis of pneumonia--a critical view].

text: OBJECTIVES To search literature related to the etiological diagnosis of acute pneumonia in children. SOURCES Systematic review of Medline and Lilacs databases. SUMMARY OF THE FINDINGS The use of new diagnostic methods such as immunological techniques and polymerase chain reaction has proven invaluable for specific diagnosis and epidemiological investigation, showing adequate sensitivity, specificity and promptness of results, with the aim of guiding therapy properly. Review of epidemiological studies of community acquired pneumonia showed that Streptococcus pneumoniae is still one of the most significant etiologic agents in all age groups, in developing and industrialized countries. Resistance of this agent to penicillin and cephalosporins is increasing in all continents and is worrisome. Atypical agents such as Mycoplasma pneumoniae and Chlamydia pneumoniae are common in community acquired pneumonia, mainly in children older than 4 years, representing one third of the cases in industrial countries. However, their prevalence in developing countries remain to be determined. Respiratory syncytial virus is also a very common etiology of community acquired pneumonia and may cause severe infections, mainly in infants and younger children. The introduction of new conjugated vaccines for Streptococcus pneumoniae and Haemophilus influenzae type b resulted in significant reduction of morbidity and mortality of pneumonia in children. CONCLUSIONS A significant impact on morbidity and mortality of acute pneumonia in children is likely to occur if microbiological and antimicrobial control is continuously and dynamically performed, thus allowing for the development of new vaccines, particularly against the respiratory syncytial virus.


title: The epidemiology of the outbreak of severe acute respiratory syndrome (SARS) in Hong Kong--what we do know and what we don't.

text: Severe acute respiratory syndrome (SARS) struck Hong Kong bitterly in the spring of 2003, infecting 1755 persons and claiming nearly 300 lives. The epidemic was introduced by travellers from southern China, where the disease had originated. It started in late February and lasted until early June. Two notable 'super-spreading' events were reported, one inside a teaching hospital and the other in a private housing estate. Other than in the super-spreading events, the infectivity in the community appeared to be low, and there were few, if any, asymptomatic or subclinical infections. Health-care workers were at particular risk and accounted for 22 % of all probable cases. The main modes of transmission were through droplet spread and close/direct contacts, but situations conducive to aerosol generation appeared to be associated with higher risk. Our review suggests that there are still many unknown factors concerning the mode of transmission and environmental risk that need to be clarified.


title: Distribution of lesions and antigen of highly pathogenic avian influenza virus A/Swan/Germany/R65/06 (H5N1) in domestic cats after presumptive infection by wild birds.

text: In early 2006, the highly pathogenic avian influenza virus (HPAIV) H5N1 of the Asian lineage caused the death of wild aquatic birds in Northern Germany. In the mainly affected areas, a trans-species transmission of HPAIV H5N1 to mammals occurred between birds and domestic cats and 1 Stone Marten (Martes foina), respectively. Here, we report lesions and distribution of influenza virus antigen in 3 cats infected naturally with HPAIV H5N1 A/swan/Germany/R65/06. The hemagglutinin partial nucleotide sequences of the viruses were genetically closely related to a H5N1 HPAIV obtained from a dead Whooper Swan (Cygnus cygnus) of the same area. At necropsy, within the patchy dark-red and consolidated lungs, there was granulomatous pneumonia caused by Aelurostrongylus sp. Histologically, the main findings associated with influenza in all cats were bronchointerstitial pneumonia and marked random hepatic necrosis. In addition, all animals displayed lymphoid necrosis in the spleen and Peyer's patches and necrosis of the adrenal cortex. Immunohistochemically, nucleoprotein of HPAIV was present intralesionally in the lungs, liver, adrenal glands, and lymphoid tissues. Oropharyngeal swabs were shown to be suited to detect HPAIV by quantitative real-time polymerase chain reaction (RT-PCR) in these cats, despite the paucity of influenza virus antigen in the upper respiratory tract by means of immunohistochemistry. The results show that outdoor cats in areas affected by HPAIV in wild birds are at risk for lethal infection. In conclusion, hepatic necrosis was, besides bronchointerstitial pneumonia, the primary lesion, suggesting that in naturally infected cats, damage to the liver plays an important role in the pathogenesis of H5N1 influenza.


title: Results of Conserve Plus Hip Resurfacing: prospective clinical, radiographic and ion study.

text: We report the 3- to 5-year clinical, radiographic and serum ion level results of a prospective consecutive cohort of 42 hip resurfacing arthroplasties using the Conserve Plus implant in 39 male patients that were operated on by a single surgeon in a community hospital. Average age was 53 years (range 34-67) at surgery. There was one revision for a subcapital neck fracture. There were no surgery related complications. The survival of the implant was 95%. Clinical evaluation showed excellent results with a modified Charnley score of 17.6/18, Harris Hip Score of 96.2/100, WOMAC of 95.1/100, Oxford Score 15.3, and UCLA-Activity Score of 8/10. Radiographic analysis showed no implant at risk, no migration or signs of loosening, no neck narrowing and no osteolysis at final follow-up. Average cup inclination angle was 43.5° with 2 outliers (34° and 57°). Ion level study showed average cobalt in serum 1.04 µg/l (range 0-4) for the whole group, 0.7 µg/l (range 0-3) in patients with unilateral resurfacing and 2.0 µg/l (range 0-4) in patients with bilateral resurfacing. All patients had ion levels within the safe zone. This independent series of Conserve Plus HRA confirms good results at short- to mid-term with excellent wear characteristics. Results for avascular necrosis were equal to those for osteoarthritis.


title: Disease contact tracing in random and clustered networks.

text: The efficacy of contact tracing, be it between individuals (e.g. sexually transmitted diseases or severe acute respiratory syndrome) or between groups of individuals (e.g. foot-and-mouth disease; FMD), is difficult to evaluate without precise knowledge of the underlying contact structure; i.e. who is connected to whom? Motivated by the 2001 FMD epidemic in the UK, we determine, using stochastic simulations and deterministic 'moment closure' models of disease transmission on networks of premises (nodes), network and disease properties that are important for contact tracing efficiency. For random networks with a high average number of connections per node, little clustering of connections and short latency periods, contact tracing is typically ineffective. In this case, isolation of infected nodes is the dominant factor in determining disease epidemic size and duration. If the latency period is longer and the average number of connections per node small, or if the network is spatially clustered, then the contact tracing performs better and an overall reduction in the proportion of nodes that are removed during an epidemic is observed.


title: [Detection of the anti-SARS-coronavirus specific antibody levels in 156 SARS patients].

text: The objective of this study was to explore the development of IgG and IgM against SARS CoV and characteristics of changes of antibody titers in patients with severe acute respiratory syndrome (SARS) and to search the opportunity for collecting specific anti-serum from convalescent patients with SARS. The anti-SARS-coronavirus specific antibody levels in 156 SARS patients were measured with ELISA. The results showed that the total positive rates of IgG and IgM were 75.6% and 41.7% respectively, and the negative rate of both IgG and IgM was 23.7%. The average titers of IgG and IgM antibody in positive samples were 18.23 +/- 24.72 and 2.18 +/- 1.13, respectively. There was no significant correlation between the titers of IgG/IgM and sex, age, course of diseases and duration of body temperature recovery. It was concluded that not all SARS patients could produce the anti-SARS-coronavirus specific antibody. The titers of the anti-body are diversified even if the antibodies have been emerged in them. In order to obtain effective anti-serum, the titers of antibody must be tested just before collection of convalescent serum, and it ensures the therapeutic effect and provides a measurable index for clinical transfusion.