title: Follow-up on diagnostic proficiency of laboratories equipped to perform orthopoxvirus detection and quantification by PCR: the second international external quality assurance study.

text: Two years after the first external quality assurance study on bioterrorism-relevant viruses, we have conducted a follow-up study on orthopoxvirus detection by PCR. Thirty-three laboratories (27 European, 4 Austral-Asian, and 2 American) participated. Samples contained 0 to 40,000,000 DNA copies of lyophilized monkeypox, cowpox, and vaccinia virus per ml. Laboratories achieved a >80% detection chance above 56,234 copies per ml. Global sensitivity was not significantly improved over that of the first study. Twenty-seven and 9 participants, respectively, were able to genotype and quantify virus. Four of 27 genotyping results were incorrect. Quantification accuracy was significantly better for vaccinia virus than for the other viruses. False-positive results occurred in 22 (11.8%) of all 186 tests on negative samples, but 18 of these were contributed by only five laboratories. Fifty-five percent of laboratories could appropriately detect PCR inhibition. The use of either real-time PCR or commercial diagnostic kits had significant positive influence on laboratory performance.


title: Outcomes of SARS survivors in China: not only physical and psychiatric co-morbidities.

text: The year 2013 marks the 10th anniversary of the outbreak of the severe acute respiratory syndrome (SARS). We present a comprehensive introduction to the current situation of surviving SARS victims in China where the disease originated and spread across the world 10 years ago.


title: [The quantitative detection of anti-coronavirus antibody titer in medical personnel closely contacted with severe acute respiratory syndrome patients].

text: OBJECTIVE To study the serum anti-coronavirus antibody titer in medical personnel who had closely contacted with severe acute respiratory syndrome (SARS) patients. METHODS The serum anti-coronavirus IgG antibody titer in medical personnel who had closely contacted with SARS patients, healthy individuals, patients with community acquired pneumonia and patients recovered from SARS was detected by using an enzyme-linked immunosorbent assay (ELISA) method. The antibody titer was expressed as the value of absorbency (A) with common logarithm conversion. RESULTS The serum anti-coronavirus IgG antibody titer in patients recovered from SARS was 0.07 +/- 0.13, which was significantly higher as compared with those in other groups. The antibody titer in medical personnel was -1.18 +/- 0.20, which was also significantly higher as compared with those in community acquired pneumonia patients and healthy persons. In the healthy persons, the antibody titer of serum samples obtained from Beijing in May, 2003 was -1.61 +/- 0.13, which was significantly higher than that of samples obtained from Beijing in 2001 when SARS was not found -1.76 +/- 0.25 and that of samples from Shandong province where SARS was not found in May, 2003 -1.95 +/- 0.44. There was no significant difference in the antibody titer between patients of bacterial pneumonia and patients of atypical pneumonia, which was -1.99 +/- 0.31 and -2.05 +/- 0.23 respectively. CONCLUSION Close contact with SARS patients can cause the serum anti-coronavirus antibody titer to increase significantly in medical personnel, a phenomenon deserves further study.


title: Genetic screen for monitoring severe acute respiratory syndrome coronavirus 3C-like protease.

text: A novel coronavirus (SCoV) is the etiological agent of severe acute respiratory syndrome. Site-specific proteolysis plays a critical role in regulating a number of cellular and viral processes. Since the main protease of SCoV, also termed 3C-like protease, is an attractive target for drug therapy, we have developed a safe, simple, and rapid genetic screen assay to monitor the activity of the SCoV 3C-like protease. This genetic system is based on the bacteriophage lambda regulatory circuit, in which the viral repressor cI is specifically cleaved to initiate the lysogenic-to-lytic switch. A specific target for the SCoV 3C-like protease, P1/P2 (SAVLQ/SGFRK), was inserted into the lambda phage cI repressor. The target specificity of the SCoV P1/P2 repressor was evaluated by coexpression of this repressor with a chemically synthesized SCoV 3C-like protease gene construct. Upon infection of Escherichia coli cells containing the two plasmids encoding the cI. SCoV P1/P2-cro and the beta-galactosidase-SCoV 3C-like protease constructs, lambda phage replicated up to 2,000-fold more efficiently than in cells that did not express the SCoV 3C-like protease. This simple and highly specific assay can be used to monitor the activity of the SCoV 3C-like protease, and it has the potential to be used for screening specific inhibitors.


title: A comparison between immunofluorescence staining on smears from Membrana nictitans (M3 test), immunohistopathology and routine pathology in cats with suspected feline infectious peritonitis (FIP).

text: An indirect immunofluorescence method using smears from membrana nictitans (M3 test) to diagnose feline corona virus (FCV) infection was compared with immunohistopathology (using indirect immunofluorescence assay (IFFA) performed on organs (IFO], and routine pathology (RP) in cats with suspected feline infectious peritonitis (FIP). A close correlation between the 2 immunofluorescence methods (IFO and M3) was observed. Although the M3 test requires samples from only 1 organ per animal, both the sensitivity and specificity were high (80%), when compared to IFO (using samples from an average of 5 organs per animal). In 21% of the cats with suspected FIP typical pathological lesions were found. As the M3 test is relatively easy to perform, it could reduce work-load of pathology laboratories and provide valuable data for clinical and epidemiological use.


title: Individual space-time activity-based modelling of infectious disease transmission within a city.

text: This paper provides an example of the application of an individual space-time activity-based model (ISTAM) to the simulation of the transmission of infectious disease in Eemnes, a city in The Netherlands. Four questions were addressed: (i) how to build the whole population at the city level, (ii) how to build the structure of the activity bundles for the city, (iii) how to assign daily activity patterns to each individual, and (iv) how to simulate within-AB transmission. The model was calibrated and examples of simulation results such as dynamics of the population during a whole day, infection distribution and network analysis are presented.


title: Detection of a novel human coronavirus by real-time reverse-transcription polymerase chain reaction.

text: We present two real-time reverse-transcription polymerase chain reaction assays for a novel human coronavirus (CoV), targeting regions upstream of the E gene (upE) or within open reading frame (ORF)1b, respectively. Sensitivity for upE is 3.4 copies per reaction (95% confidence interval (CI): 2.5–6.9 copies) or 291 copies/mL of sample. No cross-reactivity was observed with coronaviruses OC43, NL63, 229E, SARS-CoV, nor with 92 clinical specimens containing common human respiratory viruses. We recommend using upE for screening and ORF1b for confirmation.


title: M gene evolution of canine coronavirus in naturally infected dogs.

text: Two stray pups (A and B), three and five months old, respectively, both naturally infected with canine coronavirus (CCoV), were studied for 180 days. The virus was detected intermittently in the pups' faeces by PCR for periods of 156 and 146 days, respectively. Sequence analysis of a fragment of the gene encoding the M protein revealed that the viruses detected at the onset of the infection were very similar to typical strains of CCoV, whereas from 42 days after infection in pup A and 40 days after infection in pup B the viruses had nucleotide and amino acid mutations resembling sequences in feline coronavirus.


Mental health services.


title: Persistent infection promotes cross-species transmissibility of mouse hepatitis virus.

text: Persistent infection with mouse hepatitis virus (MHV) strain A59 in murine DBT (delayed brain tumor) cells resulted in the emergence of host range variants, designated V51A and V51B, at 210 days postinfection. These host range mutants replicated efficiently in normally nonpermissive Chinese hamster ovary (CHO), in human hepatocarcinoma (HepG2), and to a lesser extent in human breast carcinoma (MCF7) cell lines. Little if any replication was noted in baby hamster kidney (BHK), green African monkey kidney (COS-7), feline kidney (CRFK), and swine testicular (ST) cell lines. By fluorescent antibody (FA) staining, persistent viruses V10B and V30B, isolated at days 38 and 119 days postinfection, also demonstrated very low levels of replication in human HepG2 cells. These data suggest that persistence may rapidly select for host range expansion of animal viruses. Pretreatment of HepG2 cells with a polyclonal antibody directed against human carcinoembryonic antigens (CEA) or with some monoclonal antibodies (Col-1, Col-4, Col-12, and Col-14) that bind human CEA significantly inhibited V51B infection. Under identical conditions, little or no blockade was evident with other monoclonal antibodies (kat4c or Col-6) which also bind the human CEA glycoproteins. In addition, an antibody (EDDA) directed against irrelevant antigens did not block V51B replication. Pretreatment with the Col-4 and Col-14 antibodies did not block Sindbis virus replication in HepG2 cells or MHV infection in DBT cells, suggesting that one or more CEA glycoproteins likely functioned as receptors for V51B entry into human cell lines. To test this hypothesis, the human biliary glycoprotein (Bgp) and CEA genes were cloned and expressed in normally nonpermissive BHK cell lines by using noncytopathic Sindbis virus replicons (pSinRep19). By growth curves and FA staining, human CEA and to a much lesser extent human Bgp functioned as receptors for V51B entry. Furthermore, V51B replication was blocked with polyclonal antiserum directed against human CEA and Bgp. Under identical conditions, the parental MHV strain A59 failed to replicate in BHK cells expressing human Bgp or CEA. These data suggest that MHV persistence may promote virus cross-species transmissibility by selecting for virus variants that recognize phylogenetic homologues of the normal receptor.


Elective and emergency surgery in patients with severe acute respiratory syndrome (SARS).


title: Detection of protozoan and bacterial pathogens of public health importance in faeces of Corvus spp. (large-billed crow).

text: Parasites and bacteria are reported in the faeces of birds in the current study. Fresh faecal samples of the large-billed crow (Corvus spp.) were collected from the study site at Bangsar, an urban setting in Kuala Lumpur, Malaysia. These samples were transported to laboratory and analysed for parasites and bacteria. Pre-prepared XLD agar plates were used for culturing the bacteria in the laboratory. Using the API 20ETM Test Strips, 9 different species of bacteria were identified belonging to the family Enterobacteriacea. They were Citrobacter freundii, Enterobacter cloacae, Proteus mirabilis, Klebsiella pneumoniae, Kluyvera ascorbata, Salmonella arizonae, Salmonella typhi, Shigella flexneri and Shigella sonnei. The protozoan parasites detected include Cryptosporidium spp., Cyclospora spp., Blastocystis spp., and Capillaria hepatica and Ascaris lumbricoidus ova. Environmental air samples collected on agar plates using an air sampler in the area only produced fungal colonies. Some of these pathogens found in the crows are of zoonotic importance, especially Cryptosporidium, Blastocystis, Cyclopsora, Salmonella, Shigella and Kluyvera. The finding of Kluyvera spp. in crows in our current study highlights its zoonotic potential in an urban setting.


title: [Clinical observation on treatment of SARS with combination of chaihu droplet pill and huoxiang zhengqi droplet pill].

text: OBJECTIVE To observe the effect of combination treatment with Chaihu Droplet pill (CHDP) and Huoxiang Zhengqi Droplet pill (HZDP) in treating severe acute respiratory syndrome (SARS) in early stage. METHODS Twenty-two patients of SARS were randomly selected and paired according to such controlling factors as age, sex and profession into 11 couples. To them all symptomatic treatment was applied, combined CHDP and HZDP (CH-HZ) was given additionally to one of each couple randomly. The treatment was lasted for 13 days to investigate the changes of the clinical indexes such as creatine kinase (CK), lactate dehydogenase (LDH) and serum sodium levels. RESULTS Early applying of CH-HZ treatment showed good effects in improving CK, LDH, oxygenation index and absolute value of neutrophils, and could reduce the daily maximal dosage of glucocorticoid needed for SARS patients. CONCLUSION Early application of CH-HZ treatment in treating SARS could alleviate the injury in lung of SARS patients and the neutrophil dependent inflammatory reaction, and reduce the dosage of glucocorticoid used.


title: Sequences of the nucleocapsid genes from two strains of avian infectious bronchitis virus.

text: cDNAs prepared from viral genomic RNA purified from two strains of infectious bronchitis virus (IBV) (Beaudette and M41) have been cloned into pBR322. Three of these clones, which contain the complete sequences of mRNA A for both strains, except for the leader sequences which are only present on the subgenomic messenger RNAs, have been sequenced using the dideoxy method. The sequences are similar for both strains, each containing a single long open reading frame of 1227 bases which predicts a polypeptide of molecular weight approximately 45 000. The genome position and size of this predicted polypeptide are consistent with it being the gene for the nucleocapsid protein. The amino acid sequence shows considerable homology with those of the nucleocapsids of murine hepatitis virus strains A59 and JHM. The major difference between the sequences determined for the two IBV strains is that the 3' non-coding region of the Beaudette strain contains a 184 base segment which is not present in the M41 strain.


Apoptotic cell death during renal transplant rejection.


When a vaccine is safe.


The General Medical Council and the future of revalidation: GMC member responds.


title: Ribonucleocapsid formation of severe acute respiratory syndrome coronavirus through molecular action of the N-terminal domain of N protein.

text: Conserved among all coronaviruses are four structural proteins: the matrix (M), small envelope (E), and spike (S) proteins that are embedded in the viral membrane and the nucleocapsid phosphoprotein (N), which exists in a ribonucleoprotein complex in the lumen. The N-terminal domain of coronaviral N proteins (N-NTD) provides a scaffold for RNA binding, while the C-terminal domain (N-CTD) mainly acts as oligomerization modules during assembly. The C terminus of the N protein anchors it to the viral membrane by associating with M protein. We characterized the structures of N-NTD from severe acute respiratory syndrome coronavirus (SARS-CoV) in two crystal forms, at 1.17 A (monoclinic) and at 1.85 A (cubic), respectively, resolved by molecular replacement using the homologous avian infectious bronchitis virus (IBV) structure. Flexible loops in the solution structure of SARS-CoV N-NTD are now shown to be well ordered around the beta-sheet core. The functionally important positively charged beta-hairpin protrudes out of the core, is oriented similarly to that in the IBV N-NTD, and is involved in crystal packing in the monoclinic form. In the cubic form, the monomers form trimeric units that stack in a helical array. Comparison of crystal packing of SARS-CoV and IBV N-NTDs suggests a common mode of RNA recognition, but they probably associate differently in vivo during the formation of the ribonucleoprotein complex. Electrostatic potential distribution on the surface of homology models of related coronaviral N-NTDs suggests that they use different modes of both RNA recognition and oligomeric assembly, perhaps explaining why their nucleocapsids have different morphologies.


title: Disulfide bonds in folding and transport of mouse hepatitis coronavirus glycoproteins.

text: We have analyzed the effects of reducing conditions on the folding of the spike (S) protein and on the intracellular transport of the membrane (M) protein of the mouse hepatitis coronavirus. These proteins differ in their potential to form disulfide bonds in the lumen of the endoplasmic reticulum (ER). Intrachain disulfide bonds are formed in the S protein but not in M, which was demonstrated in a pulse-chase experiment by analyzing the viral proteins under nonreducing conditions. To reduce disulfide bonds in vivo, we added dithiothreitol (DTT) to the culture medium of mouse hepatitis coronavirus-infected cells following a procedure recently described by Braakman et al. (I. Braakman, J. Helenius, and A. Helenius, EMBO J. 11:1717-1722, 1992). Short exposure to DTT resulted in the complete reduction of newly synthesized S protein and affected its conformation as judged by the change in mobility in nonreducing gels and by the loss of recognition by a conformation-specific monoclonal antibody. Using this antibody in an immunofluorescence assay, we monitored the reducing effect of DTT in situ. DTT was found to initially affect only the S protein present in the ER; also, after longer treatment, the remaining signal also gradually disappeared. In contrast, folding and transport of the M protein were not inhibited by DTT. Under reducing conditions, M was transported efficiently to the trans side of the Golgi complex, indicating that cellular processes such as ER-to-Golgi transport, O-glycosylation, and Golgi retention were unaffected. In the presence of DTT, the M protein even moved at an increased rate to the Golgi complex, which is probably because of its failure to interact with unfolded S protein. The effects of in vivo reduction were reversible. When DTT was removed from pulse-labeled cells, the S protein folded posttranslationally and aberrantly; during its oxidation, most of S now transiently aggregated into large disulfide-linked complexes from which subsequently folded S molecules dissociated.


title: [Sequence analysis for genes encoding nucleoprotein and envelope protein of a new human coronavirus NL63 identified from a pediatric patient in Beijing by bioinformatics].

text: The aim of this study was to characterize the N and E protein encoding genes of a new human coronavirus (HCoV-NL63) which was identified from one of the clinical specimens (BJ8081) collected from a 12 years-old patient with acute respiratory infection in Beijing. The complete N and E gene sequences of HCoV-NL63 were amplified from clinical sample by RT-PCR, then were cloned into the pCF-T and pUCm-T vectors respectively and sequenced. The complete sequences of N and E genes were submitted to GenBank by Sequin and compared with N and E genes of prototype HCoV-NL63 and the other coronaviruses published in GenBank. The secondary structure and the characteristics of sample BJ8081 N and E proteins were predicted by bioinformatics. It was indicated that the N and E genes amplified from sample BJ8081 were 1134 bp and 234 bp in length and the predicted proteins including 377 amino acids and 77 amino acids, respectively. The data suggested that the region of amino acids 78-85 within N protein probably was the conserved region for all coronaviruses identified so far including HCoV-NL63. The region of amino acids 15-37 for E protein was probably the transmembrane domain. In conclusion, the recombinant plasmids pCF-T-8081 N and pUCm-T-8081 E were successfully constructed and sequenced, and the data predicted by bioinformatics are helpful for the further analysis of HCoV-NL63.


title: Changing patterns of pneumoconiosis mortality--United States, 1968-2000.

text: Pneumoconioses are caused by the inhalation and deposition of mineral dusts in the lungs, resulting in pulmonary fibrosis and other parenchymal changes. Many persons with early pneumoconiosis are asymptomatic, but advanced disease often is accompanied by disability and premature death. Known pneumoconioses include coal workers' pneumoconiosis (CWP), silicosis, asbestosis, mixed dust pneumoconiosis, graphitosis, and talcosis. No effective treatment for these diseases is available. This report describes the temporal patterns of pneumoconiosis mortality during 1968-2000, which indicates an overall decrease in pneumoconiosis mortality. However, asbestosis increased steadily and is now the most frequently recorded pneumoconiosis on death certificates. Increased awareness of this trend is needed among health-care providers, employers, workers, and public health agencies.


title: [Effectiveness of the contingency plan of the internal inflammatory disease unit before Covid-19 infection.]

text: OBJECTIVE Decision making in hospitals, and especially in their own healthcare services, is hardly referenced in the literature. During the pandemic period, healthcare services have put in place contingency plans to minimize the consequences of the coronavirus on professionals and patients. However, the deployment of contingency plans and results are hardly shared, depriving other services of references to refute, compare or emulate the aforementioned plans. The objective of this work was the description of the implementation and evaluation of Contingency Plans in the Covid-19 pandemic in a unit of inflammatory bowel disease of a Digestive Service in the Sanitary Area of Pontevedra and O Salnés. METHODS A team of managers and professionals adapted the 10 measures recommended by Deloitte to face a pandemic to the healthcare environment. The measures were then formulated as a checklist. From the Plan-Do-Check-Act improvement cycle, they were grouped into categories: risk management, organizational management and decision-making. Finally, an external team carried out a qualitative evaluation of the implementation of the contingency plan carried out. RESULTS The Intestinal Inflammatory Disease Unit of the Digestive Service has obtained an assessment of compliance with the 10 recommended measures to confidently face a pandemic. CONCLUSIONS Sharing the deployment of the contingency plan and its results is useful to identify good practices. This article shows a method to evaluate decision-making in pandemic situations. The outcomes faces the The Intestinal Inflammatory Disease Unit in an excelent position.


title: In vivo replication of porcine reproductive and respiratory syndrome virus in swine alveolar macrophages and change in the cell population in bronchoalveolar lavage fluid after infection.

text: Replication of porcine reproductive and respiratory syndrome (PRRS) virus in swine alveolar macrophages (AM) and cell population in broncho-alveolar lavage fluid (BALF) obtained from PRRS virus-infected pigs were investigated. BALF samples were periodically collected from 6 pigs infected with PRRS virus and 3 non-inoculated control pigs by means of fiber-optic bronchoscope between post-inoculation day (PID) 0 and 56. The mean ratio of macrophages in BALF collected from infected group was 92.7 +/- 3.2% before inoculation and gradually decreased from PID 14. On the other hand, the ratio of lymphocytes was 4.8 +/- 3.2% before inoculation and increased from PID 21 and indicated 41.8 +/- 9.1% on PID 28. After that, they decreased gradually and that of macrophages correspondingly increased. The ratio of neutrophils maintained between 0.7% and 5.1%. The ratios of macrophages, lymphocytes and neutrophils collected from control group were almost stable through the examination. Intracellular PRRS virus antigens in AM were detected from PID 2 by indirect immunofluorescence assay (IIFA). PRRS virus was first isolated from BALF samples collected from inoculated group between PID 2 and 49. From serum, virus was isolated between PID 2 and 21. Antibodies in sera measured by IIFA to PRRS virus were first detected on PID 14 and the antibody titer rose to 1:640 or 1:1,280. The results suggested that PRRS virus replicates in swine AM for a relatively long period.


title: Initial sequencing and tissue distribution of Toll-like receptor 3 mRNA in white-tailed deer (Odocoileus virginianus).

text: Toll-like receptor (TLR) 3 recognizes double-stranded RNA (dsRNA) and activates a signal transduction pathway that results in the release of a variety of chemokines and cytokines and apoptotic activity. Variability in TLR3 expression may play an important role in disease susceptibility of white-tailed deer (WTD; Odocoileus virginianus) to bluetongue and epizootic hemorrhagic disease viruses, which are dsRNA viruses. Because little is known about TLR3 in WTD, our objective was to sequence WTD TLR3 mRNA and to determine baseline levels of tissue expression. A 209-base pair sequence of TLR3 mRNA was obtained from WTD peripheral blood mononuclear cells. Dot blots confirmed that the sequence obtained was part of total WTD mRNA. Variable expression or ligand binding of TLR3 may contribute to observed susceptibility differences between populations of WTD, so the level of TLR3 in small intestine, skin, spleen, heart, cecum, rumen, lymph node, lung, kidney, and liver from WTD fawns (n=2) was analyzed using real-time reverse transcriptase-polymerase chain reaction. Tissue expression of TLR3 mRNA relative to the housekeeping gene beta-actin was highest in spleen, heart, skin, and lung.


title: Graphical perception and graphical methods for analyzing scientific data.

text: Graphical perception is the visual decoding of the quantitative and qualitative information encoded on graphs. Recent investigations have uncovered basic principles of human graphical perception that have important implications for the display of data. The computer graphics revolution has stimulated the invention of many graphical methods for analyzing and presenting scientific data, such as box plots, two-tiered error bars, scatterplot smoothing, dot charts, and graphing on a log base 2 scale.


Systemic vasculitis: state of the art and emerging concepts.


title: Characterization of a new species of adenovirus in falcons.

text: In 1996, a disease outbreak occurred at a captive breeding facility in Idaho, causing anorexia, dehydration, and diarrhea or sudden death in 72 of 110 Northern aplomado falcons (Falco femoralis septentrionalis) from 9 to 35 days of age and in 6 of 102 peregrine falcons (Falco peregrinus) from 14 to 25 days of age. Sixty-two Northern aplomado and six peregrine falcons died. Epidemiologic analyses indicated a point source epizootic, horizontal transmission, and increased relative risk associated with cross-species brooding of eggs. Primary lesions in affected birds were inclusion body hepatitis, splenomegaly, and enteritis. The etiology in all mortalities was determined by molecular analyses to be a new species of adenovirus distantly related to the group I avian viruses, serotypes 1 and 4, Aviadenovirus. In situ hybridization and PCR demonstrated that the virus was epitheliotropic and lymphotropic and that infection was systemic in the majority of animals. Adeno-associated virus was also detected by PCR in most affected falcons, but no other infectious agents or predisposing factors were found in any birds. Subsequent to the 1996 epizootic, a similar disease caused by the same adenovirus was found over a 5-year period in orange-breasted falcons (Falco deiroleucus), teita falcons (Falco fasciinucha), a merlin (Falco columbarius), a Vanuatu peregrine falcon (Falco peregrinus nesiotes), and gyrfalcon x peregrine falcon hybrids (Falco rusticolus/peregrinus) that died in Wyoming, Oklahoma, Minnesota, and California. These findings indicate that this newly recognized adenovirus is widespread in western and midwestern North America and can be a primary pathogen in different falcon species.


title: Nonfatal motor-vehicle animal crash-related injuries--United States, 2001-2002.

text: In 2000, an estimated 6.1 million light-vehicle (e.g., passenger cars, sport utility vehicles, vans, and pickup trucks) crashes on U.S. roadways were reported to police. Of these reported crashes, 247,000 (4.0%) involved incidents in which the motor vehicle (MV) directly hit an animal on the roadway. Each year, an estimated 200 human deaths result from crashes involving animals (i.e., deaths from a direct MV animal collision or from a crash in which a driver tried to avoid an animal and ran off the roadway). To characterize nonfatal injuries from these incidents, CDC analyzed data from the National Electronic Injury Surveillance System-All Injury Program (NEISS-AIP). This report summarizes the results of that analysis, which indicated that, during 2001-2002, an estimated 26,647 MV occupants per year were involved in crashes from encounters with animals (predominantly deer) in a roadway and treated for nonfatal injuries in U.S. hospital emergency departments (EDs). Cost-effective measures targeting both drivers (e.g., speed reduction and early warnings) and animals (e.g., fencing and underpasses) are needed to reduce injuries associated with MV collisions involving animals.


title: Human norovirus infection and the lessons from animal caliciviruses.

text: PURPOSE OF REVIEW Human noroviruses are a major cause of infectious intestinal disease, particularly in the health sector, with considerable knock-on effects on care provision through ward closures and staff sickness. This review will describe recent advances in our understanding of human noroviruses. In addition, we will consider related nonhuman caliciviruses to highlight some potential difficulties in the control of caliciviral disease. RECENT FINDINGS Using more sensitive reverse transcriptase polymerase chain reaction based assays, noroviruses are now recognized as the most common cause of infectious intestinal disease in the community, as well as outbreaks of the infectious intestinal disease. After recovery from acute disease, some individuals continue shedding norovirus, particularly if immunosuppressed. The noroviruses are extremely variable, which has important implications for protection following challenge, and for future vaccination. From amongst this variability, new strains have emerged with the potential to spread widely. Recently a mouse norovirus has been identified which will afford new insights into the biology of these important viruses. Studies on human susceptibility have identified some resistant individuals in the population and a potential virus receptor, which may lead to the development of novel antiviral therapies. SUMMARY Lack of cell culture systems for the human noroviruses is being overcome by molecular technologies. Such studies have provided new insight into the significance and epidemiology of these viruses and opened the possibility of disease control through vaccination. Work on nonhuman caliciviruses has interesting parallels with human noroviruses, and provides new insights into the understanding of these important human pathogens.


Will research sharing keep pace with the Internet?


Advertising campaign on a major internet search engine to promote colorectal cancer screening.


title: Neuronal cell surface molecules mediate specific binding to rabies virus glycoprotein expressed by a recombinant baculovirus on the surfaces of lepidopteran cells.

text: The existence of specific rabies virus (RV) glycoprotein (G) binding sites on the surfaces of neuroblastoma cells is demonstrated. Spodoptera frugiperda (Sf21) cells expressing G of the RV strain CVS (Gcvs-Sf21 cells) bind specifically to neuroblastoma cells of different species but not to any other cell type (fibroblast, myoblast, epithelial, or glioma). Attachment to mouse neuroblastoma NG108-15 cells is abolished by previous treatment of Gcvs-Sf2 cells with anti-G antibody. Substitutions for lysine at position 330 and for arginine at position 333 in RV G greatly reduce interaction between Gcvs-Sf21 cells and NG108-15 cells. These data are consistent with in vivo results: an avirulent RV mutant bearing the same double mutation is not able to infect sensory neurons or motoneurons (P. Coulon, J.-P. Ternaux, A. Flamand, and C. Tuffereau, J. Virol. 72:273-278, 1998) after intramuscular inoculation into a mouse. Furthermore, infection of NG108-15 cells by RV but not by vesicular stomatitis virus leads to a reduction of the number of binding sites at the neuronal-cell surface. Our data strongly suggest that these specific attachment sites on neuroblastoma cells represent a neuronal receptor(s) used by RV to infect certain types of neurons in vivo.


SARS. Researcher told to stay home after China trip.


title: Estimation of influenza and severe acute respiratory illness incidence (burden) in three provinces of the Islamic Republic of Iran, 2012 and 2013.

text: There are few estimates of influenza burden in the WHO Region for the Eastern Mediterranean. In this study we estimated the burden of severe acute respiratory infection (SARI) and influenza-associated SARI (F-SARI) in selected provinces of Islamic Republic of Iran, the trends of SARI and confirmed cases of influenza (F-SARI) over 12 months (seasonality), and the age groups most at risk. Using the electronic Iranian influenza surveillance system and data of cases in sentinel hospitals of 3 selected provinces, we estimated the monthly trend (seasonality) of incidence for SARI and F-SARI, overall incidence of SARI and F-SARI and their disaggregation by age with the aid using the Monte Carlo technique. The age groups most at-risk were children aged under 2 years and adults older than 50 years.


title: Epidemic dynamics on scale-free networks with piecewise linear infectivity and immunization.

text: We examine epidemic thresholds for disease spread using susceptible-infected-susceptible models on scale-free networks with variable infectivity. Infectivity between nodes is modeled as a piecewise linear function of the node degree (rather than the less realistic linear transformation considered previously). With this nonlinear infectivity, we derive conditions for the epidemic threshold to be positive. The effects of various immunization schemes including ring and targeted vaccination are studied and compared. We find that both targeted and ring immunization strategies compare favorably to a proportional scheme in terms of effectiveness.


title: The first autopsy case of pandemic influenza (A/H1N1pdm) virus infection in Japan: detection of a high copy number of the virus in type II alveolar epithelial cells by pathological and virological examination.

text: We report the pathological and virological findings of the first autopsy case of the 2009 pandemic influenza (A/H1N1pdm) virus infection in Japan. A man aged 33 years with chronic heart failure due to dilated cardiomyopathy, mild diabetes mellitus, atopic dermatitis, bronchial asthma, and obesity died of respiratory failure and multiple organ dysfunction syndrome. Macroscopic examination showed severe pulmonary edema and microscopically the lung sections showed very early exudative-stage diffuse alveolar damage (DAD). Immunohistochemistry revealed proliferation of the influenza (A/H1N1pdm) virus in alveolar epithelial cells, some of which expressed SAalpha2-3Gal on the cell surface. Influenza (A/H1N1pdm) virus genomic RNA and mRNA were also detected in alveolar epithelial cells. Real-time PCR revealed 723 copies/cell in the left lower lung section from which the influenza (A/H1N1pdm) virus was isolated. Electron microscopic analysis revealed filamentous viral particles in the lung tissue. The concentrations of various cytokines/chemokines in the serum and the autopsied lung tissue were measured. IL-2R, IL-6, IL-8, IL-10, IFN-alpha, MCP-1, and MIG levels were elevated in both. These findings indicated a case of viral pneumonia caused by influenza (A/H1N1pdm) virus infection, showing characteristic pathological findings of the early stage of DAD.


title: Information retrieval in respiratory care: tips to locate what you need to know.

text: We need current information to support effective practice. This information can be accessed relatively quickly and inexpensively, using nothing more than a computer and an Internet connection. This demands skills in searching and accessing that information. In many hospitals, computers with an Internet connection are readily available in the clinical wards, allowing those with skills in retrieving medical information to find that information near the bedside where it is needed. This report describes techniques to access information on the Internet from various sources. This information exists at high levels, such as PubMed, the National Guideline Clearinghouse, and the Cochrane Library. However, there is also much information available on the Internet that has not been validated or subjected to peer review. Thus, it is important not only to find information but also to separate useful from useless information.


Influenza-like illness and viral aetiology in Hong Kong children.


title: Breda virus (Toroviridae) infection and systemic antibody response in sentinel calves.

text: Enzyme-linked immunosorbent assays were established to detect Breda virus antigen in feces and homologous antibodies of the IgG1, IgM, and IgA isotypes in serum. With the aid of solid-phase immune-electron microscopy, torovirions in fecal material were observed. The course of natural infection was studied in 10 sentinel calves that had been obtained from different farms, and housed together at 1 week of age. They were separated from other cattle until the age of 10 months. Up to the age of 4 months, all calves regularly excreted Breda virus in the feces. Irrespective of the existence of IgG1 isotype maternal antibodies, all calves had early IgM responses in serum, but lack of IgA seroconversion. In 7 calves, antibody titer decreased below detection, whereas 3 calves had an isotype switch, resulting in persistent IgG1 titer. After introduction into the dairy herd at 10 months of age, all calves had diarrhea, and shedding of Breda virus was observed in 8 of them. Seroconversion for all antibody isotypes was observed, indicating lack of mucosal memory. In contrast, coronavirus infection in the presence of maternal antibodies led to isotype switch in all calves but one, and a memory response was observed after introduction into the dairy herd.


title: Single-payer health care systems: the roles and responsibilities of the public and private sectors.

text: Health care systems all over the world are experiencing some change as they look for a new balance between supply and demand. This article provides context for the U.S. health care financing debate by examining the health care systems of five other countries: Canada, the United Kingdom, Australia, China and India. The authors show that, with few exceptions, countries around the world have seen an increase in both government and private health care spending between 1998 and 2002. The authors also demonstrate that employers throughout most of the world are becoming more, rather than less, involved in the funding and delivery of health care to employees and their dependents-even among nations with so-called single-payer health systems.


title: Seroprevalence of infectious disease agents in free-ranging Florida panthers (Felis concolor coryi).

text: Serum samples obtained from 38 free-ranging Florida panthers (Felis concolor coryi) in southern Florida, March 1978 through February 1991, were tested for antibodies against eight bacterial, parasitic, and viral disease agents. Sera were positive for antibodies against feline panleukopenia virus (FPV) (78%), feline calicivirus (56%), feline immunodeficiency virus/puma lentivirus (37%), feline enteric coronavirus/feline infectious peritonitis virus (19%), and Toxoplasma gondii (9%). All samples were seronegative for Brucella spp., feline rhinotracheitis virus, and pseudorabies virus. In addition, all the animals tested were negative for feline leukemia virus p27 antigen as determined by enzyme-linked immunosorbent assay. Feline panleukopenia virus was considered to be a potentially significant disease agent; FPV antibodies occurred in the highest prevalences in older age classes (P = 0.027) and in panthers living in the dense mixed hardwood swamps in the western portion of their range compared to the open cypress and sawgrass prairies to the east (P = 0.096). Because < 50 animals remain in this relict population and the probable resultant depression of genetic diversity and lowered disease resistance, FPV or other disease agents could contribute to the extinction of this endangered subspecies.


title: Lessons from the response to A H1N1 influenza, 2009, India: ethics in pandemic planning.

text: After the Severe Acute Respiratory Syndrome (SARS) experience in 2003, ethics has found a place in discourses on pandemic planning and public health. It is no longer enough to merely have action strategies in a pandemic plan; both research literature and the World Health Organization recommend that one has to further ensure that the outcome of such action is fair to all concerned, and is in conformity with relevant ethical values and considerations. India's pandemic plan suffers from a glaring omission in this aspect. Taking strategies and responses during the 2009 A H1N1 outbreak in India as instances, this paper identifies the lessons to be learnt from this experience and argues that these raise ethical issues ingrained in pandemic planning which must be addressed. It ends with the suggestion that the Indian health authorities should add an ethical dimension to the national pandemic plan, as has been recommended by the World Health Organization and by post-SARS studies.


Health in a changing world.


SARS or not SARS: outbreak of fever in a state mental institute in Singapore.


Upper respiratory tract infection.


title: Decontamination of rat embryos and transfer to specific pathogen-free recipients for the production of a breeding colony.

text: When animals are introduced to a specific pathogen-free (SPF) facility, care must be taken to avoid the possibility of disease transmission to the local colony. This study investigated the application of a combination of reproductive biotechnologies to establish a new disease-free colony of two rat strains, DarkAgouti(DA/Pit) and Wistar Furth(WF/Pit), from a stock known to be chronically infected with the following pathogens: Mycoplasma pulmonis, Kilham's rat virus, sialodacryoadenitis/coronavirus, and reovirus type 3. To eliminate the pathogens and optimize the use of animals, superovulation, embryo washing and trypsinization, and embryo transfer were used. Donors (DA/Pit and WF/Pit) were treated as follows: the mature females were synchronized by subcutaneous (s.c.) injection with 40 micrograms luteinizing hormone-releasing hormone agonist/animal on day -4. All immature and mature females were induced to superovulate by continuous s.c. infusion with a commercial porcine follicle-stimulating hormone (FSH) preparation (3.4 or 6.8 mg NIH-FSH-P1 units per day, respectively), beginning on the morning of day -2. On the afternoon of day 0, the animals received 30 IU human chorionic gonadotropin injected intraperitoneally and mated. From a total of 213 ova flushed from the oviducts of 16 programmed donors, 195 transferrable two-cell embryos were recovered. Two outbred strains of SPF rats, Long-Evans (LE) and Wistar (W), were used as recipients. These mature females (LE and W) were synchronized by using luteinizing hormone-releasing hormone agonist as described and made pseudopregnant by cervical stimulation. Two-cell embryos (DA/Pit and WF/Pit) were washed and trypsinized, then transferred to the oviducts of the pseudopregnant recipients (LE and W).(ABSTRACT TRUNCATED AT 250 WORDS)


Recent advances: medical microbiology.


title: In situ data collection and structure refinement from microcapillary protein crystallization.

text: In situ X-ray data collection has the potential to eliminate the challenging task of mounting and cryocooling often fragile protein crystals, reducing a major bottleneck in the structure determination process. An apparatus used to grow protein crystals in capillaries and to compare the background X-ray scattering of the components, including thin-walled glass capillaries against Teflon, and various fluorocarbon oils against each other, is described. Using thaumatin as a test case at 1.8 Å resolution, this study demonstrates that high-resolution electron density maps and refined models can be obtained from in situ diffraction of crystals grown in microcapillaries.


title: The safety of addition of nitrous oxide to general anaesthesia in at-risk patients having major non-cardiac surgery (ENIGMA-II): a randomised, single-blind trial.

text: BACKGROUND Nitrous oxide is commonly used in general anaesthesia but concerns exist that it might increase perioperative cardiovascular risk. We aimed to gather evidence to establish whether nitrous oxide affects perioperative cardiovascular risk. METHODS We did an international, randomised, assessor-blinded trial in patients aged at least 45 years with known or suspected coronary artery disease having major non-cardiac surgery. Patients were randomly assigned via automated telephone service, stratified by site, to receive a general anaesthetic with or without nitrous oxide. Attending anaesthetists were aware of patients' group assignments, but patients and assessors were not. The primary outcome measure was a composite of death and cardiovascular complications (non-fatal myocardial infarction, stroke, pulmonary embolism, or cardiac arrest) within 30 days of surgery. Our modified intention-to-treat population included all patients randomly assigned to groups and undergoing induction of general anaesthesia for surgery. This trial is registered at ClinicalTrials.gov, number NCT00430989. FINDINGS Of 10,102 eligible patients, we enrolled 7112 patients between May 30, 2008, and Sept 28, 2013. 3543 were assigned to receive nitrous oxide and 3569 were assigned not to receive nitrous oxide. 3483 patients receiving nitrous oxide and 3509 not receiving nitrous oxide were assessed for the primary outcome. The primary outcome occurred in 283 (8%) patients receiving nitrous oxide and in 296 (8%) patients not receiving nitrous oxide (relative risk 0·96, 95% CI 0·83–1·12; p=0·64). Surgical site infection occurred in 321 (9%) patients assigned to nitrous oxide, and in 311 (9%) patients in the no-nitrous oxide group (p=0·61), and severe nausea and vomiting occurred in 506 patients (15%) assigned to nitrous oxide and 378 patients (11%) not assigned to nitrous oxide (p<0·0001). INTERPRETATION Our findings support the safety profile of nitrous oxide use in major non-cardiac surgery. Nitrous oxide did not increase the risk of death and cardiovascular complications or surgical-site infection, the emetogenic effect of nitrous oxide can be controlled with antiemetic prophylaxis, and a desired effect of reduced volatile agent use was shown. FUNDING Australian National Health and Medical Research Council; Australian and New Zealand College of Anaesthetists; Heart and Stroke Foundation of Quebec, Heart and Stroke Foundation of Ontario, Canada; General Research Fund of the Research Grant Council, Hong Kong Special Administrative Region, China.


title: Pharmacoeconomic evaluation of immunoglobulin treatment in patients with antibody deficiencies from the perspective of the German statutory health insurance.

text: Primary antibody deficiencies are the most common forms of primary immunodeficiencies. Substitution therapy with polyvalent immunoglobulins has been established as the standard therapy for antibody deficiencies for several decades. Until now mainly intravenous immunoglobulins (IVIG) have been used in Germany, and the majority of patients receive treatment in hospital outpatient clinics. In recent years subcutaneous administration of immunoglobulins (SCIG) has been developed which is administered as home self-infusion. Studies indicate no significant differences in immunoglobulin substitution therapy between SCIG and IVIG concerning outcome. We carried out a cost-minimization analysis to compare the two treatment alternatives in Germany. Under base case assumptions the treatment with SCIG is cost saving from the perspective of the German statutory health insurance. The main cost drivers are IVIG and SCIG; the incremental cost of SCIG compared to IVIG is most sensitive to changes in the immunoglobulin price and changes in the body weight of the patient.


Tuberculosis, nontuberculous lung infection, pleural disorders, pulmonary function, respiratory muscles, occupational lung disease, pulmonary infections, and social issues in AJRCCM in 2004.


title: Evolution of mouse hepatitis virus: detection and characterization of spike deletion variants during persistent infection.

text: High-frequency RNA recombination has been proposed as an important mechanism for generating viral deletion variants of murine coronavirus. Indeed, a number of variants with deletions in the spike glycoprotein have been isolated from persistently infected animals. However, the significance of generating and potentially accumulating deletion variants in the persisting viral RNA population is unclear. To study this issue, we evaluated the evolution of spike variants by examining the population of spike RNA sequences detected in the brains and spinal cords of mice inoculated with coronavirus and sacrificed at 4, 42, or 100 days postinoculation. We focused on the S1 hypervariable region since previous investigators had shown that this region is subject to recombination and deletion. RNA isolated from the brains or spinal cords of infected mice was rescued by reverse transcription-PCR, and the amplified products were cloned and used in differential colony hybridizations to identify individual isolates with deletions. We found that 11 of 20 persistently infected mice harbored spike deletion variants (SDVs), indicating that deletions are common but not required for persistent infection. To determine if a specific type of SDV accumulated during persistence, we sequenced 106 of the deletion isolates. We identified 23 distinct patterns of SDVs, including 5 double-deletion variants. Furthermore, we found that each mouse harbored distinct variants in its central nervous system (CNS), suggesting that SDVs are generated during viral replication in the CNS. Interestingly, mice with the most severe and persisting neurological disease harbored the most prevalent and diverse quasispecies of SDVs. Overall, these findings illustrate the complexity of the population of persisting viral RNAs which may contribute to chronic disease.


title: Clinical Profile of Covid-19 Infected Patients Admitted in a Tertiary Care Hospital in North India.

text: Background The novel coronavirus (Covid-19) continues to wreck havoc across China, European countries, USA and now seems to gain a strong foothold in India. The aim of this report is to describe the clinical profiles of these Covid-19 infected patients admitted in Sawai Mansingh Hospital(S.M.S), Jaipur ranging from their age, sex, travel history, clinical symptoms, laboratory evaluation, radiological characteristics, treatment provided along with common side effects and the final outcome. The described cases are one of the earliest cases of Covid-19 in the Indian subcontinent. Methods Epidemiological, clinical, laboratory, and radiological characteristics and treatment and outcomes data were obtained with data collection forms from electronic medical records and history given by 21 Covid-19 infected patients admitted in S.M.S., Jaipur. Patients were tested for Covid-19 by real-time reverse transcription polymerase chain reaction (RT-PCR) assay of 2019-nCoVRNA. Results and Discussion During the course of this study 21 Covid-19 positive patients were admitted in S.M.S Hospital, Jaipur. Male patients constituted 66.66% of total patients and majority of the patients (80.90%) were below 60 years of age. Most of the patients (71.40%) were either foreigners or had a history of foreign travel suggesting that these cases were not community acquired except for 4 cases from textile producing district Bhilwara (known as Manchester of India), an epicenter of North India. Approximately 33.33% patients were completely asymptomatic and of those who were symptomatic cough was the most common symptom (85.71%) followed by fever (78.57%), myalgia (64.28%), headache (28.57%) and dyspnea (28.57%). Three patients (14.28 %) had underlying co morbidity in the form of hypertension, diabetes mellitus, hypothyroidism, chronic kidney disease or coronary artery disease. 11 patients (52.38%) had lymphopenia in their hemogram during the course of admission. 3 patients (14.28%) had leucocytosis and 4 patients (19.04%) presented with thrombocytopenia. All 4 patients in the severe category had raised FDP, D-Dimer levels and they needed oxygen support. These patients had deranged liver functions and had elevated pro-calcitonin levels, serum ferritin levels and LDH levels. 1 out of the these 4 cases went into ARDS during the course of treatment. 10 patients yielded negative results for Covid-19. The mean duration from admission to getting 1st Covid-19 sample negative was 8.3 days. 18 patients (85.71%) are still under treatment. Conclusion Clinical investigations in initial Covid-19 patients in the Indian subcontinent reveal lymphopenia as predominant finding in hemogram. Patients with older age and associated comorbid conditions (COPD and diabetes) seem to have greater risk for lung injury thereby requiring oxygen support during the course of disease and these patients also had greater derangement in their biochemical profile.


title: [Temperature changes of patients with severe acute respiratory syndrome: a comparative study].

text: OBJECTIVE To explore the patterns of temperature changes of patients with the severe acute respiratory syndrome (SARS) and the effect of glucocorticoid hormone on the temperature of these patients. METHODS The clinical data of 94 SARS cases treated during the outbreak of SARS in South China in 2003 were collected for a retrospective review. According to different treatment regimens, the patients were divided into hormone group (n=35) and non-hormone group (n=59). The control groups consisted of 65 patients with interstitial pneumonia, 78 with bacterial pneumonia and 57 with upper respiratory tract infection. The changes in body temperature were compared between the SARS patients and those with other respiratory diseases and the effect of glucocorticoid hormone on controlling body temperature of the SARS patients was explored. RESULTS The body temperature of patients with the 4 diseases all exhibited obvious reduction 7 days after hospitalization (P<0.001) with only subsequent mild fluctuation within the basically normal range. At each time point of measurement, the body temperature of SARS patients was significantly higher than that of patients with other diseases (P<0.03), with a fluctuation of 0.2 to 0.5 degrees C; and following a pattern of variation similar to those of the other diseases. Of the 4 time points of daily measurement, namely 6, 10, 14 and 18 o'clock, the temperature measured at 14 o'clock was significantly higher than those at the other 3 time points (P<0.001). Hormone therapy did not significantly affect the temperature of SARS patients (P=0.180), who had longer duration of high fever. CONCLUSION SARS patients have higher body temperature and longer duration of high fever. Hormone therapy may not produce significant effect in controlling the temperature of SARS patients.


title: Conversion of viramidine to ribavirin in vivo by adenosine deaminase and its inhibition by 2'-deoxycoformycin.

text: Previously we reported that viramidine is a prodrug of ribavirin and that adenosine deaminase catalyses viramidine deamination to ribavirin in vivo. This in vivo study explores this prodrug conversion in rats and inhibition by a potent adenosine deaminase inhibitor, 2'-deoxycoformycin. We found that conversion of viramidine to ribavirin was viramidine dose-dependent in rat plasma. A single intravenous dose of 0.25 mg/kg 2'-deoxycoformycin suppressed orally administered viramidine conversion to ribavirin in plasma by 50%. The inhibition was 2'-deoxycoformycin dose-dependent and a single dose of 2 mg/kg decreased the ribavirin/viramidine area under the concentration-time curve between 0 h and 6 h ratio by 2.5-fold. These findings provide strong evidence that adenosine deaminase plays a major role in converting viramidine to ribavirin in vivo.


title: [The first 29 COVID-19-patients in a clinic: early experiences from a Dutch hospital].

text: The SARS-CoV-2 virus and COVID-19 disease is of pandemic proportions and reached the Netherlands on February 27 2020. Here we present the first Dutch cohort of 29 hospitalized patients during the first two weeks of the epidemic in the Netherlands. Demographic characteristics of patients, clinical presentation and course of disease up to the moment of analysis showed similarity with what has been described in Chinese and Italian literature. However the higher proportion of patients presenting with gastro-intestinal symptoms and the high number of patients with overweight and obesity stood out. Based on the experience in our hospital very early on in the epidemic COVID-19 impresses as a severe illness with risk of acute respiratory deterioration.


title: Human exposures to a rabid bat - Montana, 2008.

text: On September 29, 2008, the Ravalli County Public Health Department (RCPHD) notified the Montana Department of Public Health and Human Services (MDPHHS) of a large-scale human exposure to a dead bat at an elementary school. On October 1, the bat was confirmed to be rabid, and on October 4, MDPHHS requested assistance from CDC in evaluating persons for rabies exposure. Of 107 persons assessed, only one person (1%) was recommended for rabies postexposure prophylaxis (PEP) in accordance with guidance from the Advisory Committee on Immunization Practices (ACIP); however, 74 persons (68%) ultimately pursued rabies PEP. This report describes the incident and public health response, and highlights the importance of unified risk communication. After a potential large-scale exposure to rabies virus, guidance from clinicians should be consistent with ACIP recommendations to ensure appropriate use of rabies PEP.


title: Radiologic pattern of disease in patients with severe acute respiratory syndrome: the Toronto experience.

text: Severe acute respiratory syndrome (SARS) is a transmissible febrile respiratory illness caused by a recently discovered coronavirus. Various patterns of disease progression may be observed that have different implications for the prognosis in those affected by SARS. The appearance of the lungs on chest radiographs of patients with this condition may be normal or may include focal airspace opacity or multifocal or diffuse opacities. Thoracic computed tomography (CT) is more sensitive in depicting SARS than is conventional chest radiography, and CT images obtained in patients with normal chest radiographs may show extensive disease and airspace consolidation. However, because the radiologic appearance of SARS is not distinct from that of other diseases that cause lower respiratory tract infection, early identification of SARS will depend in part on the prompt recognition of clusters of cases of febrile respiratory tract illness. To aid in the differential diagnosis and management of SARS, radiologists must be familiar with the typical clinical and histopathologic findings, as well as the radiologic features of the disease.


title: Haemagglutination by avian infectious bronchitis virus-a coronavirus.

text: The haemagglutinating ability of three strains of IBV was investigated. It was shown that whereas strain Beaudette had no detectable haemagglutinin, both Connecticut and Massachusetts agglutinated red cells of various species. The haemagglutinin of Connecticut was detectable after sucrose gradient purification whereas that of Massachusetts required both the purification step and incubation with the enzyme phospholipase C to reveal it. The agglutination could be inhibited by specific antisera. Some studies on the nature of the red cell receptor, and the possible presence of a receptor destroying enzyme, are reported.


title: Equine viral arteritis.

text: Equine viral arteritis (EVA) can cause prominent economic losses for the equine industry. The purpose of this review is to provide the pathologist some familiarity with the clinical history, lesions, pathogenesis, and diagnosis of EVA. EVA is caused by an arterivirus (equine arteritis virus, EAV), and the vascular system is the principal but not unique viral target. EVA has variable presentations, including interstitial pneumonia, panvasculitis with edema, thrombosis and hemorrhage, lymphoid necrosis, renal tubular necrosis, abortion, and inflammation of male accessory genital glands. EAV antigen (EAVAg) can be demonstrated within the cytoplasm of epithelial cells such as alveolar pneumocytes, enterocytes, adrenal cortical cells, trophoblasts, thymus stroma, renal tubular cells, and male accessory genital gland cells. It can be also demonstrated within endothelia, in vascular, myometrial, and cardiac myocytes, macrophages, dendritelike cells of lymphoid organs, and chorionic mesenchymal stromal cells. In young and adult horses, following colonization of macrophages, the virus spreads systemically using circulating monocytes and enters the endothelium and tunica media of blood vessels, histiocytes, and dendritelike cells. Eventually, the virus multiplies within renal tubular cells. Lesions are uncommon in the aborted fetus; if present, they are mild, and EAVAg is frequently not detectable within fetal tissues and placenta. The clinical presentation and lesions of EVA may resemble those of other diseases. Complete pathologic examination associated with immunohistochemistry, virus isolation, and, especially in cases of abortion, serology will guarantee a directed and accurate diagnosis.


title: Immunodiagnosis of and immunoprophylaxis against the haemoparasites Babesia sp. and Anaplasma sp. in domestic animals.

text: The haemoparasites Babesia bovis and Anaplasma marginale are extremely important pathogens of cattle, affecting more than 300 million animals worldwide. Species of less importance affect cattle, dogs, horses and sheep. No vaccine is available for widespread usage. This review summarises existing immunodiagnostic and immunoprophylactic methods currently available for these diseases and reports on future immunoprophylactic methods.


COVID-19: The impact on pediatric emergency care


title: Fast reshaping of intensive care unit facilities in a large metropolitan hospital in Milan, Italy: facing the COVID-19 pandemic emergency

text: At the end of 2019, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak spread from China all around the world, causing thousands of deaths. In Italy, the hardest hit region was Lombardy, with the first reported case on 20 February 2020. San Raffaele Scientific Institute — a large tertiary hospital and research centre in Milan, Italy — was immediately involved in the management of the public health emergency. Since the beginning of the outbreak, the elective surgical activity of the hospital was rapidly reduced and large areas of the hospital were simultaneously reorganised to admit and assist patients with coronavirus disease 2019 (COVID-19). In addition, the hospital became the regional referral hub for cardiovascular emergencies in order to keep ensuring a high level of health care to non-COVID-19 patients in northern Italy. In a few days, a COVID-19 emergency department was created, improving the general ward capacity to a total number of 279 beds dedicated to patients with COVID-19. Moreover, the number of intensive care unit (ICU) beds was increased from 28 to 72 (54 of them dedicated to patients with COVID-19, and 18 to cardiology and cardiac surgery hub emergencies), both converting pre-existing areas and creating new high technology spaces. All the involved health care personnel were rapidly trained to use personal protection equipment and to manage this particular category of patients both in general wards and ICUs. Furthermore, besides clinical activities, continuously important research projects were carried out in order to find new strategies and more effective therapies to better face an unprecedented health emergency in Italy.


title: Emerging Issues and Future Developments in Capsule Endoscopy.

text: Capsule endoscopy (CE) has transformed from a research venture into a widely used clinical tool and the primary means for diagnosing small bowel pathology. These orally administered capsules traverse passively through the gastrointestinal tract via peristalsis and are used in the esophagus, stomach, small bowel, and colon. The primary focus of CE research in recent years has been enabling active CE manipulation and extension of the technology to therapeutic functionality; thus, widening the scope of the procedure. This review outlines clinical standards of the technology as well as recent advances in CE research. Clinical capsule applications are discussed with respect to each portion of the gastrointestinal tract. Promising research efforts are presented with an emphasis on enabling active capsule locomotion. The presented studies suggest, in particular, that the most viable solution for active capsule manipulation is actuation of a capsule via exterior permanent magnet held by a robot. Developing capsule procedures adhering to current healthcare standards, such as enabling a tool channel or irrigation in a therapeutic device, is a vital phase in the adaptation of CE in the clinical setting.


title: A loop 2 cytidine-stem 1 minor groove interaction as a positive determinant for pseudoknot-stimulated -1 ribosomal frameshifting.

text: The molecular determinants of stimulation of -1 programmed ribosomal frameshifting (-1 PRF) by RNA pseudoknots are poorly understood. Sugarcane yellow leaf virus (ScYLV) encodes a 28-nt mRNA pseudoknot that promotes -1 PRF between the P1 (protease) and P2 (polymerase) genes in plant luteoviruses. The solution structure of the ScYLV pseudoknot reveals a well ordered loop 2 (L2) that exhibits continuous stacking of A20 through C27 in the minor groove of the upper stem 1 (S1), with C25 flipped out of the triple-stranded stack. Five consecutive triple base pairs flank the helical junction where the 3' nucleotide of L2, C27, adopts a cytidine 27 N3-cytidine 14 2'-OH hydrogen bonding interaction with the C14-G7 base pair. This interaction is isosteric with the adenosine N1-2'-OH interaction in the related mRNA from beet western yellows virus (BWYV); however, the ScYLV and BWYV mRNA structures differ in their detailed L2-S1 hydrogen bonding and L2 stacking interactions. Functional analyses of ScYLV/BWYV chimeric pseudoknots reveal that the ScYLV RNA stimulates a higher level of -1 PRF (15 +/- 2%) relative to the BWYV pseudoknot (6 +/- 1%), a difference traced largely to the identity of the 3' nucleotide of L2 (C27 vs. A25 in BWYV). Strikingly, C27A ScYLV RNA is a poor frameshift stimulator (2.0%) and is destabilized by approximately 1.5 kcal x mol(-1) (pH 7.0, 37 degrees C) with respect to the wild-type pseudoknot. These studies establish that the precise network of weak interactions nearest the helical junction in structurally similar pseudoknots make an important contribution to setting the frameshift efficiency in mRNAs.


title: Household transmission of SARS, 2003.

text: BACKGROUND In the 2003 outbreak in Toronto (in Ontario, Canada) of severe acute respiratory syndrome (SARS), about 20% of cases resulted from household transmission. The purpose of our study was to determine characteristics associated with the transmission of SARS within households. METHODS A retrospective cohort of SARS-affected households was studied to determine risk factors for household transmission. Questionnaires addressed characteristics of the index case, the household and behaviours among household members. Potential risk factors for secondary transmission of infection were assessed in regression models appropriate to the outcome (secondary cases) and nonindependence of household members. RESULTS The 74 households that participated included 18 secondary cases and 158 uninfected household members in addition to the 74 index cases. The household secondary attack rate was 10.2% (95% confidence interval [CI] 6.7%-23.5%). There was a linear association between the time the index patient spent at home after symptom onset and the secondary attack rate. Infected health care workers who were index cases had lower rates of household transmission. INTERPRETATION SARS transmission in households is complex and increases with the length of time an ill person spends at home. Risk of transmission was lower when the index case was a health care worker. Rapid case identification is the public health measure most useful in minimizing exposure in the home.


title: [Detection of severe acute respiratory syndrome (SARS)-associated coronavirus RNA in autopsy tissues with in situ hybridization].

text: OBJECTIVE To explore the distribution of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) in SARS autopsy tissues at the molecular level. METHODS In situ hybridization was used to detect the expression and location of SARS-CoV RNA polymerase gene in autopsy tissues from SARS-Cov-infected subjects, including the lung, spleen, lymph nodes, pituitary, pancreas, parathyroid, adrenal glands, gastrointestinal tract, skin, brain, liver, kidney, blood vessels, striated muscles of the limbs, bone marrow, heart, ovary, uterus and testicles. RESULT SARS-CoV RNA was detected in the cytoplasm of the alveolar epithelia, infiltrating mononuclear phagocytes in the lungs, serous gland epithelium of the trachea/bronchus, monocytes in the spleen and lymph nodes, acinar cells in the pancreas, acidophilic cells in the parathyroid and pituitary, adrenal cortical cells, epithelia of the alimentary tracts, gastric parietal cells, sweat gland cells, brain neurons, hepatocytes near the central vein, epithelia of the distal renal tubules, bone marrow promyelocytes, and endothelia of the small veins. CONCLUSIONS SARS-CoV invades various organs of the body and distributes in a similar fashion to CD13, the receptor of human coronavirus 229E. The detection of SARS-CoV in the sweat glands, alimentary tracts and epithelia of the distal convoluted tubules of the kidney may help identify the transmission routes of SARS-CoV.


title: Strengthening public health laboratory capacity in Thailand for International Health Regulations (IHR) (2005).

text: INTRODUCTION Thailand conducted a national laboratory assessment of core capacities related to the International Health Regulations (IHR) (2005), and thereby established a baseline to measure future progress. The assessment was limited to public laboratories found within the Thai Bureau of Quality and Safety of Food, National Institute of Health and regional medical science centres. METHODS The World Health Organization (WHO) laboratory assessment tool was adapted to Thailand through a participatory approach. This adapted version employed a specific scoring matrix and comprised 16 modules with a quantitative output. Two teams jointly performed the on-site assessments in December 2010 over a two-week period, in 17 public health laboratories in Thailand. The assessment focused on the capacity to identify and accurately detect pathogens mentioned in Annex 2 of the IHR (2005) in a timely manner, as well as other public health priority pathogens for Thailand. RESULTS Performance of quality management, budget and finance, data management and communications was considered strong (>90%); premises quality, specimen collection, biosafety, public health functions, supplies management and equipment availability were judged as very good (>70% but ≤90%); while microbiological capacity, staffing, training and supervision, and information technology needed improvement (>60% but ≤70%). CONCLUSIONS This assessment is a major step in Thailand towards development of an optimized and standardized national laboratory network for the detection and reporting of infectious disease that would be compliant with IHR (2005). The participatory strategy employed to adapt an international tool to the Thai context can also serve as a model for use by other countries in the Region. The participatory approach probably ensured better quality and ownership of the results, while providing critical information to help decision-makers determine where best to invest finite resources.


What Have We Learned From the Covid-19 Pandemic so Far?


title: Identification, expression in E. coli and insect cells of the non-structural protein NS2 encoded by mRNA2 of bovine coronavirus (BCV).

text: The coding part of mRNA 2 (ORF2) of BCV (F15 strain) was cloned and sequenced. The comparison of our sequence data with the sequence of the same ORF of BCV Quebec strain previously published revealed a major difference in the length of the C-terminal part of the NS2 protein. In vitro transcription and translation of ORF2 resulted in the synthesis of a single protein migrating with a Mr of 31 kDa. The ORF2 was fused in frame with the glutathione S transferase gene (GSH) in the pGEX vector. The fusion protein was synthesized as inclusion bodies which were concentrated and used to raise a monospecific antiserum. Alternatively the fusion protein was solubilized, purified by affinity chromatography and cleaved with Factor Xa to yield pure recombinant NS2. The ORF2 was also expressed in the baculovirus system and the recombinant proteins expressed in pro- and eukaryotic systems were compared on the basis of their size and immunoreactivity. Immunoprecipitation performed with the monospecific antiserum allowed us to identify NS2 in HRT18 infected cells, to follow its kinetic of synthesis, and to ascertain that NS2 was not incorporated in the virion as a minor structural component.


Regulation of persistent infections with human coronavirus OC43.


title: Polyomavirus infection in a free-ranging California sea lion (Zalophus californianus) with intestinal T-cell lymphoma.

text: An adult female California sea lion (Zalophus californianus) that stranded in central California was found to have a small glossal polypoid mass on gross necropsy. Histologically, the mass was consistent with a fibropapilloma, and intranuclear inclusions were found within endothelial cells lining small arterioles within the mass. Electron microscopy revealed 40-nm virions within endothelial intranuclear inclusions. Rolling circle amplification was used to obtain a partial viral genomic sequence. Sequence analysis identified the virus as a novel polyomavirus, tentatively named California sea lion polyomavirus 1. In addition, the sea lion had a severely thickened small intestine and swollen pale kidneys on gross examination. Severe renal amyloidosis with chronic interstitial nephritis was diagnosed histologically as well as T-cell intestinal lymphoma, which was confirmed via immunophenotyping and molecular clonality. The relationship, if any, between polyomavirus infection and the other disease processes in this sea lion is not known, but it is considered unlikely that the polyomavirus induced the lymphoma.


title: Molecular determinants of human neutralizing antibodies isolated from a patient infected with Zika virus.

text: The 2015-2016 outbreak of Zika virus (ZIKV) disease has affected many countries and is a major public health concern. ZIKV is associated with fetal microcephaly and neurological complications, and countermeasures are needed to treat and prevent ZIKV infection. We report the isolation of 13 specific human monoclonal antibodies from a single patient infected with ZIKV. Two of the isolated antibodies (Z23 and Z3L1) demonstrated potent ZIKV-specific neutralization in vitro without binding or neutralizing activity against strains 1 to 4 of dengue virus, the closest relative to ZIKV. These two antibodies provided postexposure protection to mice in vivo. Structural studies revealed that Z23 and Z3L1 bound to tertiary epitopes in envelope protein domain I, II, or III, indicating potential targets for ZIKV-specific therapy. Our results suggest the potential of antibody-based therapeutics and provide a structure-based rationale for the design of future ZIKV-specific vaccines.


title: Radiographic features of SARS in paediatric patients: a review of cases in Singapore.

text: INTRODUCTION Severe acute respiratory syndrome (SARS) is a newly emerged atypical pneumonia caused by the SARS-associated coronavirus (SARS-CoV). Chest radiographic appearances have been reported as non-specific, ranging from normal to peribronchial thickening and ill-defined airspace shadowing. This study is a retrospective review of chest radiographic findings in children with suspected and probable SARS during the 2003 outbreak in Singapore. MATERIALS AND METHODS We focused on children admitted to the SARS treatment ward from March 2003 to May 2003. Chest radiographs of children admitted with suspected or probable SARS as well as other febrile illness during this period were retrospectively and independently reviewed by 3 radiologists. The radiographs were randomised and anonymised before interpretation. Subsequently, we identified the radiographs of patients who were categorised as suspected or probable SARS. We present our findings in these patients' radiographs. RESULTS A total of 67 patients' serial chest radiographs were interpreted. Of these, we subsequently selected those patients with suspected or probable SARS for analysis. The radiographic abnormalities in suspected or probable SARS patients consisted of patchy ground glass opacities or patchy airspace consolidation. The abnormalities had a predominantly lower zone distribution on chest radiographs, followed by mid-zone involvement. There was a slight preponderance of peripheral zone involvement. There was equal distribution of abnormalities in both lungs. All the children with radiographic abnormalities made uneventful recoveries and had normal radiographs on follow-up review. CONCLUSIONS In children, SARS appears to have a relatively mild and nonspecific pattern of respiratory illness. The radiographic features in children with suspected or probable SARS in our study were comparable to other clusters of paediatric patients during initial presentation. It is difficult to distinguish SARS in children from other viral pneumonias on radiographic features alone. Positive travel history to endemic regions or positive contact history, and laboratory findings of lymphopaenia, leukopaenia and thrombocytopaenia are important clues.


title: [Roles of multidisciplinary team in diagnosis and treatment of suspected cases of COVID-19].

text: OBJECTIVE To analyze the roles of multidisciplinary team (MDT) in the diagnosis and treatment of suspected cases of coronavirus disease 2019 (COVID-19). METHODS The clinical data of 48 patients with suspected COVID-19 admitted in Jinhua Municipal Central Hospital from January 21, 2020 to March 20, 2020 were retrospectively analyzed. RESULTS In the 48 suspected cases, 18 were diagnosed with COVID-19, and 30 were excluded. Each of the confirmed cases were discussed among MDT for 2 to 12 times with an average of (4.7±3.2) times; while for non-COVID-19 patients were discussed for 2 to 4 times with an average of (2.3±0.6) times. With the guidance of MDT, one COVID-19 patient was transferred to designated provincial hospital after effective treatment; one patient complicated with acute cholecystitis underwent gallbladder puncture and drainage; and COVID-19 was excluded in a highly suspected patient after alveolar lavage fluid examination. Except one transferred patient, all 17 confirmed COVID-19 patients were cured and discharged. There was no cross-infection occurred in suspected patients during the hospitalization. There were no deaths and no medical staff infections. CONCLUSIONS The efficiency of diagnosis and treatment for suspected COVID-19 patients can be improved with MDT, particularly for complicated cases.


Management of Burn Wounds Under Prolonged Field Care.


title: COVID 19- Clinical Profile, Radiological Presentation, Prognostic Predictors, Complications and Outcome: A Perspective from the Indian Subcontinent.

text: Background Since December 2019, we have been facing one of the worst pandemics of human history. It originated from the Hubei province in China as a case of pneumonia, later named COVID-19.1 The causative pathogen, a new enveloped betacoronavirus2 is now known as Severe acute respiratory syndrome corona virus-2 (SARS-CoV 2). India reported its first case of COVID19, on 30th January 2020. We aim to identify the defining clinical and radiological characteristics, severity and prognosis, along with impact of age on outcome. Methods Cross sectional, observational study of patients diagnosed with COVID -19 [RT-PCR]. Results We observed male predominance, mean age of 36 years, with less or no symptoms, majority brought in after screening and contact tracing by the screening teams. Thrombocytopenia, lymphocytosis, raised LDH was common (>35%, p<0.05). Patients over the age of 60 were the ones having severe illness and more complications (p<0.05). Radiographic abnormality was frequently associated irrespective of clinical presentation and its severity. Poor prognosis was noted in elderly, especially those with co-morbidities. Discussion Though the disease has a relatively mild course in this part of the subcontinent, patients aged ≥60 are at significant risk for morbidity and mortality. Clinical and laboratory findings are similar to those found in viral diseases. Increased risk of cardiac involvement needs to be looked into. Chest X-ray proves sufficient for imaging, reducing the requirement of CT scans. Studies involving larger sample size and interventional trials are need of the hour.


title: Neurobiological and psychological correlates of suicidal attempts and thoughts of death in patients with major depression.

text: Suicide is a major problem for psychiatry. Depression is the most common mental disorder related to suicidal behavior. The present study aimed at investigating the relationship between the symptomatology related to death, dying, and suicide and neurobiological factors in depressed patients. Fifty patients aged 21-60 years suffering from major depression were investigated. Schedules for Clinical Assessment in Neuropsychiatry version 2.0 and the International Personality Disorder Examination were used to assist the clinical diagnosis. The psychometric assessment included the Hamilton Depression Rating Scale, the Hamilton Anxiety Scale, the 1965 and 1971 Newcastle Depression Diagnostic Scales, the Diagnostic Melancholia Scale, the General Assessment of Functioning Scale, and the Personality Deviance Scale. Psychophysiological methods included electro-oculogram, flash electroretinogram under photopic and scotopic conditions, and pattern-reversal visual evoked potentials. Biological markers included the 1-mg dexamethasone suppression test, the 30-mg dexfenfluramine challenge test, and brain (99m)Tc-HMPAO SPECT. Statistical analysis included one-, two-, and three-way Manova and Mancova and the Scheffé test as post hoc test. Patients without thoughts of death had higher self-confidence levels and less overdependency on others and intropunitiveness. The suicidal patients had a significantly prolonged pattern-reversal visual evoked potential latency in comparison with the other patients. The findings of this were related to the status of the patient at the time of the interview but not to his/her history. They also provide neurobiological data to support the need for a combined presence of self-directed aggression and a higher arousal level or disinhibition of self-directed aggressive thoughts in order for a patient to become suicidal. Further study is needed to test whether psychophysiological methods, which are noninvasive and easy to perform, are of value in the therapeutic planning and monitoring of responses.


title: Antigenic variation among transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus strains detected with monoclonal antibodies to the S protein of TGEV.

text: Five nonneutralizing monoclonal antibodies (MAb) generated to the virulent Miller strain of transmissible gastroenteritis virus (TGEV) and specific for the S protein were characterized. Competition assays between purified and biotinylated MAb indicated that MAb 75B10 and 8G11 mapped near a new subsite, designated V and 2 MAb, 44C11 and 45A8, mapped to a previously designated subsite D. A fifth MAb mapped between subsites V and E. These MAb were tested with 3 previously characterized MAb to subsites A, E, and F in fixed-cell ELISA and cell culture immunofluorescent assays against 5 reference and 9 field strains of TGEV and 2 US strains (ISU-1 and ISU-3) of porcine respiratory coronavirus (PRCV). Subsites A, E, and F were conserved on all TGEV and PRCV strains examined. The 2 MAb to subsite V, 8G11 and 75B10, reacted only with the Miller TGEV strains (M5C, M6, and M60), except that 75B10 also recognized field strain U328. The MAb 11H8 did not react with 4 field strains or the Purdue strains of TGEV. The 2 MAb to subsite D reacted with all TGEV strains examined, but not with 2 US PRCV strains, 2 European PRCV strains, 1 feline infectious peritonitis virus strain, and 1 canine coronavirus strain. Because of this specificity for TGEV, but not PRCV, these latter 2 subsite D MAb may be useful for the development of competition ELISA to differentiate serologically between TGEV and PRCV infections in swine, similar to the currently used European subsite D MAb.


title: Biophysical properties of coronavirus strain OC 43.

text: Coronavirus OC 43 from suckling mouse brains was subjected to sucrose density gradient centrifugation. Maxima of complement fixing (CF) activities occurred in the ribosomal (density 1.14) and mitochondrial (density 1.19) fraction which contained incomplete, damaged virus particles. The haemagglutinating (HA) activity was associated with the microsomal soluble protein fraction displayed no antigenic activity. The isolated purified virus proved to be very unstable.


title: Detection of rotavirus and coronavirus shedding in two beef cow herds in Idaho.

text: Fecal samples were taken at the time of pregnancy examinations and at parturition from two beef herds. They were also taken from sick calves at the onset of disease, and from 25% of the healthy calves at 15 days of age. All fecal samples were examined by electron microscopy for viruses.Four cows in herd A were detected excreting coronavirus, one at the time of the pregnancy examinations and three at parturition. The first cow was removed from the herd and the others calved at the end of the season. There were no sick calves.No cows in herd B were detected excreting virus at the time of pregnancy checks, but fourteen coronavirus and two rotavirus carrier cows were found at parturition. All but two calves sampled had large numbers of virus particles in their feces. Clinical illness was associated with dams shedding virus and with nightly low temperatures.


title: Pilot survey of the diagnosis and management of heart failure in primary care: a Singaporean perspective.

text: INTRODUCTION The diagnosis and management of congestive heart failure at the primary care level is difficult. Despite advances in medical therapy, namely angiotensin-converting enzyme inhibitors and beta-blockers, congestive heart failure remains a common cause of hospital visits. The extent to which these advances have been implemented in primary care is uncertain. We therefore sought to assess current practice patterns of primary care physicians and identify obstacles to optimal diagnosis and management of congestive heart failure in Singapore. MATERIALS AND METHODS Telephone interviews of 100 primary care physicians using structured questionnaires. RESULTS Primary care physicians diagnose congestive heart failure based on the presence of breathlessness (92.5%), lung crackles (52.5%), peripheral oedema (55.0%) and findings on chest radiography (76.8%). Only 1 in 3 mentioned raised jugular venous pressure as a useful diagnostic sign. Echocardiography was not commonly obtained (5%), mainly due to inaccessibility. While many (47.5%) were aware of the importance of angiotensin-converting enzyme inhibitors, few (16.2%) knew of the impact of beta-blockers on morbidity and mortality. Instead, diuretics were thought to be most important for prolonging survival in congestive heart failure (47.5%). The main obstacle to widespread angiotensin-converting enzyme inhibitors usage was dry cough. Patient compliance (57.5%) and cost (82.5%) were the main obstacles to optimising care. CONCLUSIONS Local primary care physicians diagnose congestive heart failure using non-specific symptoms and signs. Obstacles to optimal congestive heart failure management were identified. We hope our results will lead to strategies to improve patient care.


title: Rhinoviruses and coronaviruses - virological aspects of their role in causing colds in man.

text: Rhinoviruses are picornaviruses that cause colds. They are naturally temperature sensitive and highly adapted to grow in respiratory epithelium. Coronaviruses are structurally quite different, being enveloped viruses but are also adapted to grow in respiratory epithelium and also cause colds.


Transvenous Salvage of Migrated Large Volume Detachable Coil.


title: Dodging the CTL response: viral evasion of Fas and granzyme induced apoptosis.

text: The importance of CTL induced apoptosis as a vital part of the protection of host organisms from pathogenic viruses cannot be overstated. Conversely, the ability of a virus to evade CTL induced apoptosis is equally important to its survival. Important insights in viral pathogenesis and host immunology have been discovered through observations of this constantly evolving interchange. This mini review will build upon previously published comprehensive reviews by reorganizing the anti-apoptotic strategies specific for CTL induced apoptosis and integrating recent discoveries in viral evasion of Fas/FasL and perforin/granzyme mediated apoptosis. This updated look at viral evasion in the context of the CTL response should generate dialogue and provide impetus for research to illuminate interactions between the best defense against viruses and the viral adaptations to evade this defense.


Experimental infection with a virulent pneumoenteric isolate of bovine coronavirus.


title: Risk factors for feline infectious peritonitis among cats in multiple-cat environments with endemic feline enteric coronavirus.

text: OBJECTIVE To determine what risk factors, other than genetic predisposition, contribute to the incidence of feline infectious peritonitis (FIP) in private breeding catteries and animal shelters. DESIGN Cats from 7 catteries and a shelter were observed monthly for 1 year. At each visit, cats were examined, fecal samples were collected for determination of feline coronavirus shedding, and blood samples were collected for determination of coronavirus antibody titers. Diagnostic tests were performed on all cats that died of FIP. ANIMALS 275 purebred or random-bred cats that were kept by private breeder-owners in homes. RESULTS 24 cats died of FIP during the study. Development of FIP was not associated with cattery, mean cat number, mean age, sex, cattery median coronavirus antibody titer, husbandry and quarantine practices, caging and breeding practices, or prevalence of concurrent diseases. However, risk factors for FIP included individual cat age individual cat coronavirus titer, overall frequency of fecal coronarvirus shedding, and the proportion of cats in the cattery that were chronic coronavirus shedders. Deaths from FIP were more frequent in fall and winter, and on the basis of analysis of cattery records, the number of deaths varied yearly. Epidemics (> 10% mortality rate) were reported at least once in 5 years in 4 catteries. CLINICAL IMPLICATIONS Elimination of FIP from a cattery is only possible by total elimination of endemic feline enteric coronavirus (FECV) infection. The most important procedure to reduce FECV from catteries is elimination of chronic FECV shedders.


title: Detection of the ORF3 polypeptide of feline calicivirus in infected cells and evidence for its expression from a single, functionally bicistronic, subgenomic mRNA.

text: Feline calicivirus (FCV) is a small positive-stranded RNA virus within the family Caliciviridae. Its genome is 7690 nucleotides in length and encodes three open reading frames (ORFs). The smallest, ORF3, is located at the extreme 3' end of the genome and can potentially encode a polypeptide of approximately 12 kDa. In this paper, we report the identification of an ORF3-encoded polypeptide in FCV-infected cells using an antiserum raised against a bacterially-expressed bacteriophage T7 gene 10-ORF3 fusion protein. Although a small mRNA of 0-5 kb, which could potentially encode ORF3, has been described, reports on the number and size of FCV subgenomic RNAs have varied considerably. To clarify the situation, RNAs from FCV-infected cells were labelled in vivo using [32P]orthophosphate, an approach which provided definitive data. Only two RNA species were detected, the genomic RNA and a subgenomic mRNA of 2.4 kb. The 5' end of the subgenomic mRNA was mapped to position 5227 on the genomic RNA using RNA sequencing and primer extension methods. RNA isolated from FCV-infected cells in which no subgenomic RNA smaller than 2.4 kb was detectable directed the synthesis in rabbit reticulocyte lysate of the ORF3-encoded polypeptide. Furthermore, a synthetic RNA copy of the 2-4 kb subgenomic mRNA of FCV, containing both ORF2 and ORF3 polypeptides in the in vitro translation system. These data strongly suggest that ORF3 is expressed from the 2-4 kb subgenomic RNA and that this RNA is functionally bicistronic. The possible mechanisms by which ORF3 is expressed are discussed.


title: The nucleoprotein is required for efficient coronavirus genome replication.

text: The construction of a set of transmissible gastroenteritis coronavirus (TGEV)-derived replicons as bacterial artificial chromosomes is reported. These replicons were generated by sequential deletion of nonessential genes for virus replication, using a modified TGEV full-length cDNA clone containing unique restriction sites between each pair of consecutive genes. Efficient activity of TGEV replicons was associated with the presence of the nucleoprotein provided either in cis or in trans. TGEV replicons were functional in several cell lines, including the human cell line 293T, in which no or very low cytopathic effect was observed, and expressed high amounts of heterologous protein.


title: Development of an enzyme-linked immunosorbent assay to detect chicken parvovirus-specific antibodies.

text: Here we report the development and application of an enzyme-linked immunosorbent assay (ELISA) to detect parvovirus-specific antibodies in chicken sera. We used an approach previously described for other parvoviruses to clone and express viral structural proteins in insect cells from recombinant baculovirus vectors. In baculovirus recombinant-infected Sf9 cells, the chicken parvovirus (ChPV) structural viral protein 2 (VP2) was detected as an abundant protein, and the 60-kDa VP2 strongly reacted with parvovirus-infected chicken serum in Western blot. A semipurified VP2 was then used in capture ELISA. Sera from chickens experimentally infected with ChPV and sera from uninfected chickens were tested to evaluate the assay. The ELISA was 93.3% sensitive and 100% specific in detecting ChPV-infected birds. Subsequent assays identified IgG type ChPV-specific maternally acquired antibodies in day-old chickens and demonstrated the production of virus-specific antibodies in young birds following infection with ChPV. In our study, a specific antibody response of infected chickens was observed starting with IgM production between 14 and 21 days postinfection (DPI) and switching into a predominant IgG response by 32 DPI. The availability of an ELISA for detection of virus-specific antibodies and its ability to differentiate between maternally acquired antibodies and antibodies produced following acute infection could prove to be a valuable tool to characterize pathobiological properties and immunogenicity of ChPV.


title: Detection of antibody to turkey coronavirus by antibody-capture enzyme-linked immunosorbent assay utilizing infectious bronchitis virus antigen.

text: An antibody-capture enzyme-linked immunosorbent assay (ELISA) for detection of antibody to turkey coronavirus (TCV) utilizing infectious bronchitis virus (IBV) antigen was developed. Anti-TCV hyperimmune turkey serum and normal turkey serum were used as positive or negative control serum for optimization of the ELISA system. Goat anti-turkey immunoglobulin G (light plus heavy chains) conjugated with horseradish peroxidase was used as detector antibody. The performance of the ELISA system was evaluated with 45 normal turkey sera and 325 turkey sera from the field and the cutoff point was determined. Serum samples of turkeys experimentally infected with TCV collected sequentially from 1 to 63 days postinfection were applied to the established antibody-capture ELISA using IBV antigens. The optimum conditions for differentiation between anti-TCV hyperimmune serum and normal turkey serum were serum dilution at 1:40 and conjugate dilution at 1:1600. Of the 325 sera from the field, 175 were positive for TCV by immunofluorescent antibody (IFA) assay. The sensitivity and specificity of the ELISA relative to IFA test were 93.1% and 96.7%, respectively, based on the results of serum samples from the field turkey flocks using the optimum cutoff point of 0.18 as determined by the logistic regression method. The ELISA values of all 45 normal turkey sera were completely separated from that of IFA-positive sera. The ELISA results of serum samples collected from turkeys experimentally infected with TCV were comparable to that of the IFA assay. Reactivity of anti-rotavirus, anti-reovirus, anti-adenovirus, or anti-enterovirus antibodies with the IBV antigens coated in the commercially available ELISA plates coated with IBV antigens could be utilized for detection of antibodies to TCV in antibody-capture ELISA.