title: Direct template matching reveals a host subcellular membrane gyroid cubic structure that is associated with SARS virus.

text: Viral infection can result in alterations to the host subcellular membrane. This is often reported when using transmission electron microscopy (TEM), resulting in a description of tubuloreticular membrane subcellular ultrastructure rather than a definition based on 3-D morphology. 2-D TEM micrographs depicting subcellular membrane changes are associated with subcellular SARS virion particles [Goldsmith CS, Tatti KM, Ksiazek TG et al. Ultra-structural characterization of SARS coronavirus. Emerg Infect Dis 2004; 10: 320-326]. In the present study, we have defined the 2-D membrane pattern and shape associated with the SARS virus infection. This is by using a direct template matching method to determine what the 3-D structure of the SARS virus associated host membrane change would be. The TEM image for our purposes is defined on 2-D information, such as the membrane having undergone proliferation and from pattern recognition suggesting that the membrane-described pattern is possibly a gyroid type of membrane. Features of the membrane were used to compute and match the gyroid structure with an existing 2-D TEM micrograph, where it was revealed that the membrane structure was indeed a gyroid-based cubic membrane. The 2-D gyroid computer-simulated image that was used to match the electron micrograph of interest was derived from a mathematically well-defined 3-D structure, and it is from this 3-D derivative that allows us to make inferences about the 3-D structure of this membrane. In conclusion, we demonstrate that a 3-D structure can be defined from a 2-D membrane patterned image and that a SARS viral associated membrane change has been identified as cubic membrane morphology. Possible mechanisms for this cubic membrane change are discussed with respect to viral severity, persistence and free radical production.


title: Bovine coronavirus (BCV) infections in transported commingled beef cattle and sole-source ranch calves.

text: This study investigated bovine coronavirus (BCV) in both beef calves direct from the ranch and commingled, mixed-source calves obtained from an auction market. The level of BCV-neutralizing antibodies found in the calves varied among ranches in 2 different studies in a retained-ownership program (ROP), from the ranch to the feedlot. Calves with low levels of BCV-neutralizing antibodies (16 or less) were more likely to be treated for bovine respiratory disease (BRD) than those with higher titers. In 3 studies of commingled, mixed-source calves, BCV was recovered from calves at entry to the feedlot and the infections were cleared by day 8. The BCV was identified in lung samples [bronchoalveolar lavage (BAL) collection] as well as in nasal swabs. Calves with low levels of BCV-neutralizing antibodies at entry were most likely to be shedding BCV. Bovine coronavirus was isolated from both healthy and sick calves, but not from sick calves after 4 d arrival at the feedlot. Bovine coronavirus (BCV) should be considered along with other bovine respiratory viruses in the diagnosis of etiologies in bovine respiratory disease, especially for animals that become sick shortly after arrival. If approved vaccines are developed, it would be best to carry out vaccination programs before calves are weaned, giving them sufficient time to gain active immunity before commingling with other cattle.


The SARS epidemic in Hong Kong: what lessons have we learned?


title: Are specialized endotracheal tubes and heat-and-moisture exchangers cost-effective in preventing ventilator associated pneumonia?

text: Ventilator-associated pneumonia (VAP) is a common and serious complication of mechanical ventilation via an artificial airway. As with all nosocomial infections, VAP increases costs, morbidity, and mortality in the intensive care unit (ICU). VAP prevention is a multifaceted priority of the intensive care team, and can include the use of specialized artificial airways and heat-and-moisture exchangers (HME). Substantial evidence supports the use of endotracheal tubes (ETTs) that allow subglottic suctioning; silver-coated and antiseptic-impregnated ETTs; ETTs with thin-walled polyurethane cuffs; and HMEs, but these devices also can have adverse effects. Controversy still exists regarding the evidence, cost-effectiveness, and disadvantages and risks of these devices.


title: A cross-sectional, comparative study of pain and activity in persons with and without injection-related venous ulcers.

text: Persons with leg ulcers, including venous ulcers, often report pain. A cross-sectional, comparative study was conducted among 61 patients receiving care in an urban clinic (31 with and 30 without a venous ulcer, mean age 54 years [range 40 to 65 years], 93% African American) to examine pain and its relation to activity and walking in persons with injectionrelated venous ulcers. The questionnaire included items about pain and its treatment, as well as activity and walking (ie, Brief Pain Inventory [BPI] Short Form, Self-Treatment of Pain, Pain and Narcotic Use, Difficulty with Activities, and Walking Scale questionnaires). Among those with a venous ulcer (VU+), worst pain significantly related to total interference (r = 0.65, P <0.0001) and total difficulty (r = 0.42, P = 0.02) BPI scores. The common pain sites for those VU+ involved the legs (24, 36.4%), wound sites (13, 19.7%), back (eight, 12.1%), general body (five, 7.6%), shoulder and knee (four, 6.1% each), and other (eight, 12.1%). Persons VU+ were more likely than those without a venous ulcer (VU-) to have received a prescription for narcotics in the past year (96% versus 41%, X(2) = 21.3, P <0.0001). Persons VU+ versus VU- were significantly (X(2) = 8.89, P = 0.003) more likely to resort to street drug use and relapse to addiction if pain was not adequately treated. They were also twice as likely to have decreased walking over the past 5 years (67% versus 33%, X(2) = 5.93, P <0.02). Among those VU+, venous ulcers added to chronic pain and decreased walking. These findings highlight the negative effects of injection-related venous ulcers on pain, activity, and walking, as well as the propensity of this groupto resort to illicit drug use for pain control. Persons VU+ should have pain assessed and treated.


title: Comparative analysis of the base composition and codon usages in fourteen mycobacteriophage genomes.

text: To study the possible codon usage and base composition variation in the bacteriophages, fourteen mycobacteriophages were used as a model system here and both the parameters in all these phages and their plating bacteria, M. smegmatis had been determined and compared. As all the organisms are GC-rich, the GC contents at third codon positions were found in fact higher than the second codon positions as well as the first + second codon positions in all the organisms indicating that directional mutational pressure is strongly operative at the synonymous third codon positions. Nc plot indicates that codon usage variation in all these organisms are governed by the forces other than compositional constraints. Correspondence analysis suggests that: (i) there are codon usage variation among the genes and genomes of the fourteen mycobacteriophages and M. smegmatis, i.e., codon usage patterns in the mycobacteriophages is phage-specific but not the M. smegmatis-specific; (ii) synonymous codon usage patterns of Barnyard, Che8, Che9d, and Omega are more similar than the rest mycobacteriophages and M. smegmatis; (iii) codon usage bias in the mycobacteriophages are mainly determined by mutational pressure; and (iv) the genes of comparatively GC rich genomes are more biased than the GC poor genomes. Translational selection in determining the codon usage variation in highly expressed genes can be invoked from the predominant occurrences of C ending codons in the highly expressed genes. Cluster analysis based on codon usage data also shows that there are two distinct branches for the fourteen mycobacteriophages and there is codon usage variation even among the phages of each branch.


title: Clinical manifestations and inflammatory cytokine responses in patients with severe acute respiratory syndrome.

text: BACKGROUND AND PURPOSE Severe acute respiratory syndrome (SARS) is a highly transmissible disease with significant morbidity and mortality. Death from SARS is most often due to rapidly progressive respiratory compromise (acute respiratory distress syndrome, ARDS) and subsequent multi-organ dysfunction. However, the mechanisms evoking respiratory distress and a fulminant systemic response remain unclear. In order to elucidate the pathogenic mechanisms of SARS, we analyzed clinical manifestations and levels of serum cytokines of SARS patients. METHODS Fourteen hospitalized patients with a diagnosis of SARS-associated coronavirus infection at National Taiwan University Hospital from March to May 2003 were included. Data on clinical manifestations, parameters of laboratory tests, complications and final outcomes of patients were collected retrospectively. Serial plasma inflammatory cytokines, including interleukin (IL)-1beta (IL-1beta), IL-6, IL-8 and tumor necrosis factor-alpha (TNF-alpha) of preserved serum were measured by enzyme immunoassay. RESULTS All 14 patients had fever, dry cough and dyspnea. Twelve were intubated during hospitalization. The median duration from onset of fever to the nadir level or most severe condition was 9 days for hypoxia, 7 days for lymphocytopenia, 6.5 days for thrombocytopenia, 9.5 days for maximal pulmonary infiltrates; to peak serum levels was 9 days for C-reactive protein (CRP), 10.5 days for IL-6, 13.5 days for IL-8 and 12 days for TNF-alpha; to defervescence was 13 days. There was no significant elevation of serum IL-1beta levels in any of the 14 patients. There were no significant differences in peak levels of IL-6, IL-8 and TNF-alpha between patients with and without ARDS. The 8 patients who died tended to have higher peak levels of serum TNF-alpha compared to those who survived (14 vs 9.1 pg/mL; p = 0.06). CONCLUSION Rapid elevation of inflammatory cytokines-IL-6, IL-8 and TNF-alpha might play a role in the development of SARS-related ARDS. The timing of elevations in inflammatory cytokines and CRP is correlated with progression of pulmonary infiltrates of SARS patients. The peak level of serum TNF-alpha tends to be higher in patients who die of SARS than in those who survive. Our results indicate that CRP and TNF-alpha might be used as prognostic markers of SARS.


title: Surveillance of resistance to adamantanes among influenza A(H3N2) and A(H1N1) viruses isolated worldwide.

text: Our previous reports demonstrated an alarming increase in resistance to adamantanes among influenza A(H3N2) viruses isolated in 2001-2005. To continue monitoring drug resistance, we conducted a comprehensive analysis of influenza A(H3N2) and A(H1N1) viruses isolated globally in 2005-2006. The results obtained by pyrosequencing indicate that 96.4% (n=761) of A(H3N2) viruses circulating in the United States were adamantane resistant. Drug resistance has reached 100% among isolates from some Asian countries. Analysis of correlation between the appearance of drug resistance and the evolutionary pathway of the hemagglutinin (HA) gene suggests at least 2 separate introductions of resistance into circulating populations that gave rise to identifiable subclades. It also indicates that resistant A(H3N2) viruses may have emerged in Asia in late 2001. Among A(H1N1) viruses isolated worldwide, resistance reached 15.5% in 2005-2006; in the United States alone, it was 4.0%. Phylogenetic analysis of the HA and M genes indicates that the acquisition of resistance in A(H1N1) viruses can be linked to a specific genetic group and was not a result of reassortment between A(H3N2) and A(H1N1) viruses. The results of the study highlight the necessity of close monitoring of resistance to existing antivirals as wells as the need for new therapeutics.


title: Infection of SDAV-immune rats with SDAV and rat coronavirus.

text: Infection of rats with sialodacryoadenitis virus (SDAV) or rat coronavirus (RCV) is acute and self-limiting, and elimination and control of either virus is based on the assumption that recovered rats are immune to reinfection. To test this hypothesis, we examined whether SDAV-immune rats could be infected with RCV or reinfected with SDAV. Sprague Dawley (SD) rats were inoculated intranasally with SDAV or with culture medium alone and serial SDAV antibody titers were obtained. Eleven months after inoculation, when antibody titers had stabilized, SDAV-immune and nonimmune rats were challenged with SDAV or RCV, and euthanatized 3 or 6 days later. SDAV-immune rats challenged with SDAV or RCV manifested acute rhinitis associated with virus antigen by 3 days after inoculation, but no lesions or antigen were subsequently found in the lower respiratory tract, salivary glands or lacrimal glands. There was also a marked anamnestic increase in antibody titer by 6 days after challenge. SDAV-immune rats challenged with SDAV or RCV also transmitted infection to nonimmune cage mates. This study indicates that 11 months after primary infection with SDAV, rats can be infected with SDAV or RCV, but that the severity of disease is significantly reduced.


Clostridium difficile-associated disease in the elderly, United States.


title: The effect of oversewing the staple line in laparoscopic sleeve gastrectomy: randomized control trial.

text: INTRODUCTION Laparoscopic sleeve gastrectomy (LSG) is one of the most popular bariatric procedures. The procedure is associated with serious staple-line complications such as bleeding, leaks, and stenosis. AIM To determine whether oversewing the staple line, compared with clipping, in LSG reduces the incidence of postoperative bleeding. MATERIAL AND METHODS We conducted a parallel-group, prospective, randomized controlled trial (RCT) of 100 patients who underwent LSG for obesity at a single institution between May 2014 and August 2015. Patients were assigned to one of two groups for reinforcement of the staple line: the oversewing group (staple line oversewn) or the clipping group (staple line clipped). The primary outcome was reoperation for hemodynamic instability caused by staple-line bleeding within 72 h postoperatively. The secondary outcomes were operative duration, length of hospital stay, postoperative leaks, and postoperative stenosis. RESULTS Mean operative duration was longer in the oversewing group (78.2 ±20.5 min) than in the clipping group (64.1 ±16.5 min, p < 0.001). Mean length of hospital stay was comparable in both groups. Postoperatively, there was no significant between-group difference in bleeding (oversewing, n = 0 vs. clipping, n = 2 (4.6%); p = 0.21) or in stenosis and leakage (both outcomes: oversewing, n = 0, vs. clipping, n = 1 (2.3%); p = 0.46). CONCLUSIONS Oversewing the staple line prolongs operative duration. No conclusions can be drawn regarding the effects of oversewing on staple-line bleeding, postoperative leakage and stenosis, or length of hospital stay.


title: Acute exacerbations of chronic obstructive pulmonary disease: causes and impacts.

text: Acute exacerbations of chronic obstructive pulmonary disease (AECOPD) are recognised clinically as episodes of increased breathlessness and productive cough requiring a more intensive treatment. A subset of patients with this disease is especially prone to such exacerbations. These patients are labelled as 'frequent exacerbators'. Though yet poorly characterised in terms of host characteristics, including any genetic basis, these patients are believed to represent a distinct phenotype as they have a different natural history with a more progressive disease and a poorer prognosis than those who get exacerbations infrequently. Most exacerbations appear to be associated with infective triggers, either bacterial or viral, although 'non-infective' agents, such as air pollution and other irritants may also be important. Susceptibility to exacerbations is determined by multiple factors. Several risk factors have been identified, some of which are modifiable. Chronic obstructive pulmonary disease (COPD) exacerbations are major drivers of health status and patient-centered outcomes, and are a major reason for health care utilisation including hospitalisations and intensive care admissions. These are associated with considerable morbidity and mortality, both immediate and long-term. These episodes have a negative impact on the patient and the disease including high economic burden, increased mortality, worsening of health status, limitation of activity, and aggravation of comorbidities including cardiovascular disease, osteoporosis and neuro-psychiatric complications. Exacerbations also increase the rate of progression of disease, increasing the annual decline in lung function and leading to a poorer prognosis. Evaluation of risk of exacerbations is now included as a major component of the initial assessment of a patient with COPD in addition to the traditionally used lung function parameter, forced expiratory volume in one second (FEV1). Decreasing the risk of exacerbations and their prevention is a major therapeutic goal of management in COPD.


title: [Pathological and ultramicrostructural changes of tissues in a patient with severe acute respiratory syndrome].

text: OBJECTIVE To study the morphological, ultramicrostructural and pathological changes of tissues from a patient with severe acute respiratory syndrome (SARS). METHODS One autopsy case of diagnosed SARS was investigated. Lung puncture was performed immediately after the patient died, and the autopsy was done after 12 h. The specimens from lymph nodes, spleen, small intestine, colon and bone marrow were studied by immunohistochemical technique. The antibodies used included CD20, CD45RO (UCHL-1), CD4, CD8, CD68 and CD34. RESULTS The principal lesions of the SARS case consisted of acute lobular intrastitial pneumonia, hyaloid membranes of pulmonic alveoli and hyperplasia and shedding of alveolar epithelium of. Virus-like inclusions occasionally contained cytoplasm of the alveolar epithelium, which were positive by histochemical staining. The adjacent blood-vessels were changed by hyperplasia and enlargement. The structures of lymph nodes and spleen were damaged with lymph follicles depletion and splenic nodules atrophy. The specific changes included reduction of lymphocytes and hyperplasia of histiocytes, depletion of the follicles of small intestine and colon wall, decreased hyperplasia of the bone marrow and increased number of the megakaryocyte. Meanwhile, in the immunohistochemical study, CD(20)(+) B cells were fully expressed in lymph nodes and spleen, and the CD45RO (UCHL-1)(+) T cells were scatteredly expressed. The number of CD4(+) help T cell was markedly decreased, while the number of CD8+ poisonal T cells increased, and the ratio of the former and latter was no more than 0.5. Under the electronic microscopy observation, virus-like particles with 80 - 160 nm diameter and halo or garland envelope were found in mononuclear macrophage and cytoplasm of alveolar epithelium. CONCLUSION The specific lesions of SARS consist of lobular intrastitial pneumonia with the formation of hyaline membranes of lung, haemorrhage, necrosis, inflammation of blood vessels and the damages of extralung lymphohemopioetic system. The damages were very similar to the pathological features of tissues infected by human immunodeficiency virus, in which numbers of T cells decreased and CD(4)(+) T cell/CD(8)(+) T cell ratio was no more than 0.5. According to the virus-like particles found in lung of the SARS case, it is considered that these virus-like particles may be a new kind of coronavirus which caused the "atypical pneumonia".


title: Pulmonary artery thrombosis in a patient with severe acute respiratory syndrome.

text: Severe acute respiratory syndrome (SARS) is an emerging infectious disease with both pulmonary and extra-pulmonary manifestations. Although coagulation abnormalities are common in these patients, clinically overt thromboembolic events are rarely reported. This report describes the first case of pulmonary artery thrombosis in a patient with laboratory confirmed SARS.


title: Peripheral thyroid dysfunction in depression.

text: The involvement of the thyroid gland and thyroid hormones is generally believed to be important in the aetiopathogenesis of major depression. Major support comes from studies in which alterations in components of the hypothalamic-pituitary-thyroid (HPT) axis have been documented in patients with primary depression. However, screening thyroid tests are often routine and add little to the diagnostic evaluation. Overt thyroid disease is rare among depressed inpatients. The finding that depression often co-exists with autoimmune subclinical thyroiditis suggests that depression may cause alterations in the immune system, or that in fact it could be an autoimmune disorder itself. The outcome of treatment and the course of depression may be related to thyroid status as well. Augmentation of antidepressant therapy with the co-administration of thyroid hormones (mainly T3) is a well-documented treatment option for refractory depressed patients. Review of the literature suggests that there are no conclusive data on the role of thyroid function in depression. It is clear that depression is not characterised by an overt thyroid dysfunction, but it is also clear that a subgroup of depressed patients may manifest subtle thyroid abnormalities, or an activation of an autoimmune process. There is a strong possibility that the presence of a subtle thyroid dysfunction is a negative prognostic factor for depression and may demand specific therapeutic intervention.


title: Genetic and antigenic analysis of the G attachment protein of bovine respiratory syncytial virus strains.

text: Antigenic and genetic studies of bovine respiratory syncytial virus (BRSV) were made on isolates obtained from three continents over 27 years. Antigenic variation between eight isolates was initially determined using protein G-specific monoclonal antibodies. Four distinct reaction patterns were observed, two of which corresponded to the previously established subgroups A and AB. A third pattern was produced by five Scandinavian strains and a fourth was observed from a single Dutch isolate. The genetic diversity of 27 strains of BRSV was investigated by comparative nucleotide sequence analysis of a 731 nucleotide fragment in the G protein gene. Nine of the BRSV strains were analysed by direct sequencing of RT-PCR amplicons whereas sequences of 18 BRSV and three human respiratory syncytial virus (HRSV) strains were obtained from GenBank. The analysis revealed similarities of 88-100% among BRSV strains and 38-41% between BRSV and HRSV. A phylogenetic tree created for BRSV revealed two main branches, one of which divided into five further lineages, each representing a geographic cluster. A correlation was evident between the positions of some strains in the phylogenetic tree and their antigenic pattern. For HRSV strains, a genetic similarity of only 62% allowed the distinction of two antigenic subgroups, A and B, a pattern which was not seen for BRSV. This study showed that genetic analysis was an accurate method for discriminating BRSV strains and that these viruses should be regarded as a single genetic and antigenic group, within which variants can be distinguished.


title: Characterization of Whitney's Clethrionomy gapperi virus isolates from Massachusetts.

text: Six strains of virus were recovered from the blood and/or liver of five Clethrionomys gapperi ochraceus trapped in southeastern Massachusetts during 1969. Biological, antigenic and physiochemical properties of these isolates are reported. USA M-2268a was selected as the reference strain. This strain was identical by complement-fixation and neutralization tests to Whitneys C. gapperie virus (USA 64-7855) from New York State and was related to, but distinct from, an unpublished agent (Johnson's Microtus montanus enterovirus USA M-1146) isolated in June, 1962 from voles trapped in Klamath County, Oregon. USA M-2268a was resistant to lipid solvents and acid pH and was stable at temperatures of 4 C, 22 C, and 37 C. Virus was detected over a 10-day observation period in four species of mosquitoes inoculated with USA M-2268a, although there was no evidence of infection or replication, and transmission attempts by bite failed. Neutralizing antibody was detected in C.g. gapperi and C. g. ochraceus in various habitats throughout the state.


title: Infectious disease and the decline of Steller sea lions (Eumetopias jubatus) in Alaska, USA: insights from serologic data.

text: Serologic data were examined to determine whether infectious disease may have played a role in the decline of Steller sea lions (Eumetopias jubatus) in the Gulf of Alaska and Aleutian Islands, USA. Available published data, unpublished data, and recent collections (1997-2000) were compared and reviewed. Data were stratified by geography to compare the declining western Alaskan population in the Aleutian Islands through eastern Prince William Sound to the increasing population in southeastern Alaska. Prevalences of antibodies from the 1970s to the early 1990s were noted for Leptospira interrogans, Chlamydophila psittaci, Brucella spp., phocid herpesvirus-1, and calciviruses. Serum samples collected from 1997-2000 were tested for antibodies to these agents as well as to marine mammal morbilliviruses, canine parvovirus, and canine adenovirus-1 and -2. Conclusions could not be drawn about changes in antibody prevalence to these agents during the decline of Steller sea lions, however, because data were incomplete or not comparable as a result of inconsistencies in testing techniques. Despite these shortcomings, results provided no convincing evidence of significant exposure of Steller sea lions to morbilliviruses, Brucella spp., canine parvovirus, or L. interrogans. Steller sea lions have been exposed to phocid herpesviruses, caliciviruses, canine adenovirus, and C. psittaci or to cross-reactive organisms in regions of both increasing and decreasing sea lion abundance. Based on similar antibody prevalence estimates from the increasing and decreasing populations, these agents are unlikely to have been the primary cause of the population decline. They may have contributed to the decline or impeded population recovery, however, because of undetected mortality and morbidity or reductions of fecundity and body condition in animals under other stresses. Systematic monitoring for disease agents and their effects is needed to determine whether infectious disease currently plays a role in the decline and lack of recovery of Steller sea lions.


title: Immobilization of swift foxes with ketamine hydrochloride-xylazine hydrochloride.

text: There is an increasing need to develop field immobilization techniques that allow researchers to handle safely swift foxes (Vulpes velox) with minimal risk of stress or injury. We immobilized captive swift foxes to determine the safety and effectiveness of ketamine hydrochloride and xylazine hydrochloride at different dosages. We attempted to determine appropriate dosages to immobilize swift foxes for an adequate field-handling period based on three anesthesia intervals (induction period, immobilization period, and recovery period) and physiologic responses (rectal temperature, respiration rate, and heart rate). Between October 1998-July 1999, we conducted four trials, evaluating three different dosage ratios of ketamine and xylazine (2.27:1.2, 5.68:1.2, and 11.4:1.2 mg/kg ketamine:mg/kg xylazine, respectively), followed by a fourth trial with a higher dosage at the median ratio (11.4 mg/kg ketamine: 2.4 mg/kg xylazine). We found little difference in induction and recovery periods among trials 1-3, but immobilization time increased with increasing dosage (P < 0.08). Both the immobilization period and recovery period increased in trial 4 compared with trials 1-3 (P < or = 0.03). There was a high variation in responses of individual foxes across trials, making it difficult to identify an appropriate dosage for field handling. Heart rate and respiration rates were depressed but all physiologic measures remained within normal parameters established for domestic canids. We recommend a dosage ratio of 10 mg/kg ketamine to 1 mg/kg xylazine to immobilize swift foxes for field handling.


title: Translation from the 5' untranslated region (UTR) of mRNA 1 is repressed, but that from the 5' UTR of mRNA 7 is stimulated in coronavirus-infected cells.

text: Viral gene products are generally required in widely differing amounts for successful virus growth and assembly. For coronaviruses, regulation of transcription is a major contributor to these differences, but regulation of translation may also be important. Here, we examine the possibility that the 5' untranslated regions (UTRs), unique for each of the nine species of mRNA in the bovine coronavirus and ranging in length from 70 nucleotides (nt) to 210 nt (inclusive of the common 5'-terminal 65-nt leader), can differentially affect the rate of protein accumulation. When the natural 77-nt 5' UTR on synthetic transcripts of mRNA 7 (mRNA for N and I proteins) was replaced with the 210-nt 5' UTR from mRNA 1 (genomic RNA, mRNA for viral polymerase), approximately twofold-less N, or (N) CAT fusion reporter protein, was made in vitro. Twofold less was also made in vivo in uninfected cells when a T7 RNA polymerase-driven transient-transfection system was used. In coronavirus-infected cells, this difference surprisingly became 12-fold as the result of both a stimulated translation from the 77-nt 5' UTR and a repression of translation from the 210-nt 5' UTR. These results reveal that a differential 5' UTR-directed regulation of translation can occur in coronavirus-infected cells and lead us to postulate that the direction and degree of regulation is carried out by viral or virally induced cellular factors acting in trans on cis-acting elements within the 5' UTR.


title: Detection of respiratory pathogens in air samples from acutely infected pigs.

text: Pathogens causing significant respiratory disease in growing pigs include Porcine reproductive and respiratory syndrome virus, Porcine circovirus 2, swine influenza virus, porcine respiratory coronavirus, Mycoplasma hyopneumoniae, and Bordetella bronchiseptica. The objective of this research was to characterize the respiratory excretion of these pathogens by acutely infected pigs. Pigs were inoculated under experimental conditions with 1 pathogen. Samples were collected from the upper respiratory tract and exhaled air. All pathogens were detected in swabs of the upper respiratory tract, but only M. hyopneumoniae and B. bronchiseptica were detected in expired air from individually sampled, acutely infected pigs. These findings suggest either that the acutely infected pigs did not aerosolize the viruses or that the quantity of virus excreted was below the detection threshold of current sampling or assay systems, or both, at the individual-pig level.


title: Pathogenesis of primary respiratory disease induced by isolates from a new genetic cluster of bovine viral diarrhea virus type I.

text: The pathogenesis of infection induced by cytopathogenic isolates from the newly identified genetic cluster Id of bovine viral diarrhea virus (BVDV) type I was studied in two experimental infections of previously seronegative, immunocompetent calves. Experiment 1 focused on the evaluation of clinical patterns, viremia, and serological responses. All infected calves in this experiment developed respiratory symptoms and seroconverted to BVDV positivity. Contact calves also contracted a respiratory tract infection following exposure to infected animals. Viremia was demonstrated between postinfection days 2 and 17, and the virus was detected in organ specimens of all but one each of the infected and contact calves. In experiment 2, the distribution of BVDV in various tissues of calves euthanized at defined days postinfection was studied. In two of these calves recurrent shedding of BVDV in nasal secretions was shown. BVDV was detected in various tissues of all infected calves throughout the experiment and also following seroconversion and the clearance of BVDV from the circulatory system. Despite the widespread distribution of the virus in various organs, significant tissue damage was found mainly in respiratory tract and lymphoid tissues. These experiments revealed that viruses from cluster Id of BVDV are able to induce primary respiratory disease in previously seronegative, immunocompetent calves. Contact transmission and virus recurrence, contrary to observations from acute experimental infections with noncytopathogenic BVDV, are likely to reflect differences in biological features of these cytopathogenic isolates. Virus shedding and its presence in tissues following peripheral clearance and in the presence of antibodies may have implications in the diagnosis, pathogenesis, and epidemiology of BVDV-induced syndromes in cattle.


title: [Respiratory complaints in the time of COVID-19].

text: The current guidelines for diagnosis and treatment of primary respiratory infections are still useful during the COVID-19 epidemic.Telephone triage of patients with respiratory complaints aims to identify patients with complications or an increased risk of complications.There are no indications to test for SARS-CoV-2 virus in general practice.During this COVID-19 epidemic, protective clothing is recommended in all physical contacts with patients with respiratory complaints.There is no reason to be cautious about using NSAIDs in patients suspected of COVID-19.Amoxicillin is first choice treatment for respiratory infections during the COVID-19 epidemic; there is lack of evidence to support azithromycin as a first choice.Respiratory rate > 24 / min or saturation <92-94% indicate imminent respiratory decompensation and may be reasons for referral.


title: Stereotactic radiofrequency ablation of unresectable intrahepatic cholangiocarcinomas: a retrospective study.

text: PURPOSE To evaluate treatment effects, complications, and outcome of percutaneous stereotactic radiofrequency ablation (SRFA) of intrahepatic cholangiocarcinoma (ICC). PATIENTS AND METHODS Eleven consecutive patients (nine men and two women) with a total of 36 inoperable ICCs (18 initial lesions, 16 lesions newly detected during follow-up, and two local recurrences) underwent SRFA between December 2004 and June 2010. Two different radiofrequency ablation (RFA) devices with internally cooled electrodes were used. Tumor diameters ranged from 0.5 to 10 cm (median 3.0 cm). A total of 23 SRFA sessions were performed. The efficacy of SRFA was evaluated by contrast-enhanced computed tomography or magnetic resonance imaging 1 month after treatment and then every 3 months. RESULTS Primary technical effectiveness rate was 92%. Further follow-up every 3 months revealed three local recurrences (8%), two of which were successfully retreated, resulting in a secondary technical effectiveness rate of 98%. After a total of 23 RFA sessions, three major complications occurred (13%) that could be managed interventionally. Mean follow-up time was 35 months (range 12-81 months). One- and 3-year overall survival rates were 91 and 71%, respectively. The median overall survival was 60 months (according to the life table method). Eight (73%) of 11 patients were still alive at the end of follow-up. CONCLUSION SRFA is effective in the treatment of unresectable ICC even if the tumor is large and located close to major vessels. SRFA shows a survival benefit compared to other palliative treatment options and may also be considered as the first-line local treatment of ICCs in selected patients.


title: Effect of various pyrimidines possessing the 1-[(2-hydroxy-1-(hydroxymethyl)ethoxy)methyl] moiety, able to mimic natural 2'-deoxyribose, on wild-type and mutant hepatitis B virus replication.

text: Hepatitis B virus (HBV) is the most common cause of chronic liver disease worldwide. Development of drug resistance against clinical anti-HBV drug lamivudine due to long-term use and rebound of viral DNA after cessation of treatment has been a major setback of the current therapy. We have synthesized a series of pyrimidine nucleosides possessing a variety of substituents at the C-5 position, and a 1-[(2-hydroxy-1-(hydroxymethyl)ethoxy)methyl] flexible acyclic glycosyl moiety at the N-1 position, that have the ability to mimic the natural 2'-deoxyribosyl moiety. Some of these potential antiviral compounds included variations at both C-5 and C-6 positions of the uracil base. Other variations of the uracil derivatives were the 6-aza congeners. 4-Amino and 4-methoxy pyrimidine derivatives were also made. Compounds in which the base moiety was substituted by 5-chloro- (25), 5-(2-bromovinyl)- (32), or 5-bromo-6-methyl- (37) groups possess significant activity against duck-HBV, wild-type human HBV (2.2.15 cells), and lamivudine-resistant HBV containing single and double mutations. No cytotoxicity was seen in host HepG2 and Vero cells, up to the highest concentration tested. The anti-HBV activity exhibited by compounds 25, 32, and 37 was superior for human HBV and comparable for DHBV to that of the corresponding purine nucleoside, ganciclovir. Further, they were only 10-15-fold less inhibitory against human HBV in 2.2.15 cells than the reference drug, lamivudine. Other compounds in the series were moderately inhibitory against DHBV and wild-type human HBV. The size of the halogen and the electronegativity of the substituents at the 5- and 6-positions are important for antiviral activity toward HBV. These compounds were also evaluated for their antiviral activity for West Nile virus, respiratory syncytial virus, SARS-coronavirus, and hepatitis C virus. They were generally inactive in these antiviral assay systems (at concentrations up to 100 microg/mL). 1-[(2-Hydroxy-1-(hydroxymethyl) ethoxy)methyl]-5-fluorocytosine (34) showed some inhibitory activity against hepatitis C virus. Taken together, these data support our previous observations that the 5-substituted pyrimidine nucleosides containing acyclic glycosyl moieties have potential to serve as a new generation of potent, selective, and nontoxic anti-HBV agents for wild-type and lamivudine-resistant mutant HBV.


title: Augmented reality assisted laparoscopic partial nephrectomy.

text: Computer assisted navigation is a widely adopted technique in neurosurgery and orthopedics. However, it is rarely used for surgeries on abdominal organs. In this paper, we propose a novel, noninvasive method based on electromagnetic tracking to determine the pose of the kidney. As a clinical use case, we show a complete surgical navigation system for augmented reality assisted laparoscopic partial nephrectomy. Experiments were performed ex vivo on pig kidneys and the evaluation showed an excellent augmented reality alignment error of 2.1 mm ± 1.2 mm.


[Diagnosis and treatment of acute bronchitis].


title: [Seromonitoring of laboratory mouse and rat colonies for common murine pathogens (author's transl)].

text: During a period from 1973 to 1978, 392 and 225 lots including 12,232 mouse and 8,044 rat individual sera, respectively, were examined for antibodies to murine hepatitis virus, Sendai virus, Bordetella bronchiseptica, Mycoplasma pulmonis, Tyzzer agents, Salmonella typhimurium and Corynebacterium kutscheri. Of mouse lots 94.5% and 39.3% from breeder and user colonies, respectively, were negative for all antibodies examined as well as 31.6% and 17.2% of rat breeder and user colonies, respectively. Among positive lots from mouse users, high positivity rates were seen with Senai virus (47.6%), M. pulmonis (19.0%), and murine hepatitis virus (JHM : 18.2%, MHV : 31.0%), while the rates were high in rat user lots with Sendai virus (24.4%), B. bronchiseptica (39.3%) M. pulmonis (12.5%), murine coronaviruses (JHM : 19.0%, MHV-2 : 28.0%) and tyzzer agents (MSK : 19.6%, RT : 17.9%). These pathogenes with high positivities should be monitored indispensably as a quality control of laboratory mice and rats.


title: The diverse biological roles of mammalian PARPS, a small but powerful family of poly-ADP-ribose polymerases.

text: Poly-ADP-ribose metabolism plays a mayor role in a wide range of biological processes, such as maintenance of genomic stability, transcriptional regulation, energy metabolism and cell death. Poly-ADP-ribose polymerases (PARPs) are an ancient family of enzymes, as evidenced by the poly-ADP-ribosylating activities reported in dinoflagellates and archaebacteria and by the identification of Parp-like genes in eubacterial and archaeabacterial genomes. Six genes encoding "bona fide" PARP enzymes have been identified in mammalians: PARP1, PARP2, PARP3, PARP4/vPARP, PARP5/Tankyrases-1 and PARP6/Tankyrases-2. The best studied of these enzymes PARP1 plays a primary role in the process of poly-ADP-ribosylation. PARP1-mediated poly-ADP-ribosylation has been implicated in the pathogenesis of cancer, inflammatory and neurodegenerative disorders. This review will summarize the novel findings and concepts for PARP enzymes and their poly-ADP-ribosylation activity in the regulation of physiological and pathophysiological processes. A special focus is placed on the proposed molecular mechanisms involved in these processes, such as signaling, regulation of telomere dynamics, remodeling of chromatin structure and transcriptional regulation. A potential functional cross talk between PARP family members and other NAD+-consuming enzymes is discussed.


Doubtful duty: physicians' legal obligation to treat during an epidemic.


title: Dividing the Emergency Department into Red, Yellow, and Green Zones to Control COVID-19 Infection; a Letter to Editor.

text: COVID-19, in certain respects, can be viewed as a CBRN (chemical, biological, radiological, or nuclear) event due to being a consequence of SARS-CoV2 virus (the "contaminant"). We, thus, reorganized our emergency department (ED) into 3 distinct zones (red, yellow, and green) for the purpose of infection control. Patients with high or medium risk of COVID-19 infection are managed in the red zones. Low-risk patients are managed in the yellow zones. All patients are prohibited to enter the green zones. Green zones are used by healthcare providers (HCPs) for personal protective equipment (PPE) donning, inventory, planning, and dining. Only HCPs who work in the red zones are required to use full level PPE (aerosol precaution). HCPs working in the yellow zones require less PPE (contact and droplet precaution). No PPE is required in the green zones. Establishing red, yellow, and green zones in the ED can be helpful in reducing cross-infections and minimizing demand for PPE.


title: Retrospective study of noroviruses in samples of diarrhoea from cattle, using the Veterinary Laboratories Agency's Farmfile database.

text: A collaborative study was undertaken by the Veterinary Laboratories Agency (vla) and the Royal Veterinary College (rvc) to determine the prevalence of bovine noroviruses in cattle with diarrhoea. Samples of bovine diarrhoea were provided by the vla from routine diagnostic submissions and a reverse transcription-pcr was used by the rvc to detect the viruses. Epidemiological information about the samples was provided retrospectively by the Farmfile database. Noroviruses were detected in 44 (11 per cent) of the 398 samples tested, and Farmfile data were used to investigate the differences between the positive and negative animals.


Early intervention for first episode psychosis.


title: Hand-assisted, conventional and laparoendoscopic single-site surgery for partial nephrectomy without ischemia using a microwave tissue coagulator.

text: PURPOSE We report our experience of minimally invasive partial nephrectomy without ischemia using a microwave tissue coagulator (MTC) for hand-assisted laparoscopic partial nephrectomy (HALPN), conventional laparoscopic partial nephrectomy (CLPN), and laparoendoscopic single-site surgery for partial nephrectomy (LESSPN). We retrospectively compared the results of these techniques to better define the individual role and the benefits. MATERIALS AND METHODS From July 2005 to September 2012, 28 patients with small and exophytic renal tumors underwent HALPN (n = 12), CLPN (n = 10) and LESSPN (n = 6). In these procedures, the surgeon used an MTC for circumferential coagulation around the tumor. After coagulation, the tumor was resected without renal pedicle clamping. RESULTS The mean operative time was 259, 194 and 174 min for the HALPN, CLPN and LESSPN groups respectively. Two patients (one in HALPN group and one in LESSPN group) converted to laparotomy due to an inability to maintain hemostasis; however, there were no conversions to ischemic partial nephrectomy or radical nephrectomy. No differences between HALPN, CLPN and LESSPN were noted in terms of estimated blood loss, measured analgesic requirements, outcomes, or complications. CONCLUSION We believe that these techniques are feasible and that they minimize the risk of unexpected collateral thermal damage by appropriate MTC needle puncture. When deciding to use HALPN, CLPN or LESSPN, our findings suggest that the choice of surgical approach should depend on the patient's individual circumstance.


title: Demographic effects of canine parvovirus on a free-ranging wolf population over 30 years.

text: We followed the course of canine parvovirus (CPV) antibody prevalence in a subpopulation of wolves (Canis lupus) in northeastern Minnesota from 1973, when antibodies were first detected, through 2004. Annual early pup survival was reduced by 70%, and wolf population change was related to CPV antibody prevalence. In the greater Minnesota population of 3,000 wolves, pup survival was reduced by 40-60%. This reduction limited the Minnesota wolf population rate of increase to about 4% per year compared with increases of 16-58% in other populations. Because it is young wolves that disperse, reduced pup survival may have caused reduced dispersal and reduced recolonization of new range in Minnesota.


title: Escherichia coli heat-stable enterotoxin in feces and intestines of calves with diarrhea.

text: Two experiments were conducted to evaluate detection of Escherichia coli heat-stable enterotoxin (ST) in the feces of calves as a method for implicating E coli in neonatal calf diarrhea. The first experiment evaluated the use of the infant mouse test for detection of ST in the feces of calves with naturally occurring diarrhea. Simultaneous identification of bovine enteropathogenic strains of E coli (EEC) and of other infective agents implicated in neonatal calf diarrhea was attempted in these samples. The ST was detected with certainty in only 7 of 41 samples from calves less than or equal to 3 weeks old. Enteropathogenic E coli, however, was detected in 27 samples. In 23 of these 27 samples, EEC was the only recognizable diarrheagenic agent. In a small percentage of the samples, Salmonella, rotavirus, coronavirus, and cryptosporidium were recognized alone, in combination with each other, or with EEC. In the second experiment, 6 calves were fed colostrum from cows inoculated with the bovine EEC strain B44; 6 were given colostrum from cows vaccinated with non-EEC strain 28F, and 4 were given milk from nonvaccinated heifers. Two of the calves that were given colostrum from cows inoculated with strain B44 were challenge exposed with the non-EEC strain 28F. The remaining calves were challenge exposed with the EEc strain B44. Fecal samples were taken from these calves at intervals and were examined for the presence of ST and of the challenge-exposure organism. The ST was detected in approximately one half of the fecal samples obtained, and it was most often detected in the early stages of the induced diarrhea. Calves were observed to shed the challenge-exposure EEC strain for long periods in the absence of diarrhea or detectable amounts of ST in the feces. The ST was detectable in fecal samples when the diarrhea was severe and when the dry matter content of the fecal samples was low.


title: Outbreak of severe acute respiratory syndrome in a tertiary hospital in Singapore, linked to an index patient with atypical presentation: epidemiological study.

text: OBJECTIVE To describe an outbreak of severe acute respiratory syndrome (SARS) in a tertiary hospital in Singapore, linked to an index patient with atypical presentation, and the lessons learnt from it. DESIGN Descriptive study. SETTING A tertiary hospital in Singapore. PARTICIPANTS Patients, healthcare workers, and visitors who contracted SARS in Singapore General Hospital. MAIN OUTCOME MEASURES Probable SARS as defined by the World Health Organization. RESULTS The index patient presented with gastrointestinal bleeding, initially without changes to his chest radiograph. Altogether 24 healthcare workers, 15 patients, and 12 family members and visitors were infected. The incubation period ranged from three to eight days. Only 13 patients were isolated on their dates of onset. CONCLUSIONS Atypical presentation of SARS infection must be taken into consideration when managing patients with a history of contact with SARS patients. The main gap in the containment strategy in this outbreak was the failure to identify the index patient as someone who had been discharged from a ward in another hospital that managed probable SARS cases. Strict infection control measures, a good surveillance system, early introduction of isolation procedures, and vigilant healthcare professionals are essential for controlling outbreaks.


Predicting failure of acute stroke intervention.


SARS outbreak. Modelers struggle to grasp epidemic's potential scope.


title: [Transanal endoscopic microsurgery].

text: Transanal endoscopic microsurgery was introduced by Buess and co-workers in 1984 for transanal removal of tumours up to 20 cm above anus. In the present study, 18 transmural and two mucosal resections were performed for rectal adenomas (16) and early rectal cancer (4). Median operation time was 85 minutes (55-140 minutes) and the median postoperative stay in hospital was 3 days (1-5). All tumours were removed radically and in none was there tumour involvement of the resection margin. There were no serious complications. After a median observation time of 10 months (1-17 months) there have been no recurrences. Functional results are excellent; none of the patients have developed incontinence. It is concluded that transanal endoscopic microsurgery (TEM) is an adequate method for removal of benign rectal tumours and, in selected cases, early rectal cancer.


Assembly of mouse hepatitis virus strain JHM.


title: Field investigations of emergency department syndromic surveillance signals--New York City.

text: INTRODUCTION The New York City (NYC) Department of Health and Mental Hygiene (DOHMH) has operated a syndromic surveillance system based on emergency department (ED) chief-complaint data since November 2001. This system was created for early detection of infectious-disease outbreaks, either natural or intentional. However, limited documentation exists regarding epidemiologic field investigations conducted in response to syndromic surveillance signals. OBJECTIVE DOHMH conducted field investigations to characterize syndromic surveillance signals by person, place, and time and to determine whether signals represented true infectious-disease outbreaks. METHODS A DOHMH physician reviews ED-based syndromic surveillance results daily to look for signals. When necessary, field investigations are conducted and consist of a review of the patient line list, telephone interviews with hospital staff, chart reviews, interviews with patients, and collection and testing of specimens. RESULTS In November 2002, a series of citywide signals for diarrhea and vomiting syndromes, which coincided with institutional outbreaks consistent with viral gastroenteritis, prompted DOHMH to send mass e-mail notification to NYC ED directors and institute collection of stool specimens. Three of four specimens collected were positive for norovirus. In December 2002, DOHMH investigated why an ED syndromic signal was not generated after 15 ill patients were transferred to a participating ED during a gastrointestinal outbreak at a nursing home. Field investigation revealed varying chief complaints, multiple dates of ED visits, and a coding error in a complementary DOHMH syndromic system, and confirmed a seasonal norovirus outbreak. During March 2003, the system generated a 4-day citywide respiratory signal and a simultaneous 1-day hospital-level fever signal in a predominantly Asian community. In those instances, epidemiologic investigation provided reassurance that severe acute respiratory syndrome was not present. CONCLUSION Detailed field investigations of syndromic signals can identify the etiology of signals and determine why a given syndromic surveillance system failed to detect an outbreak captured through traditional surveillance. Validation of the utility of syndromic surveillance to detect infectious-disease outbreaks is necessary to justify allocating resources for this new public health tool.


title: [Severe acute respiratory syndrome (SARS). Lesson and challenges].

text: After four months of silence before a new incursion of severe acute respiratory syndrome virus (SARS), world wide investigators specially from Asia had have to face a coronavirus capable of great spreading and inducing high morbidity. The source and the capacity of this virus to associate with other opportunist microorganisms to induce progression of the disease, remain unclear. The comparison of the genomic sequence of different strains has generated discrepancies as for if the SARS epidemic arose as an unique outbreak or it was produced for more than one genotype. However, this fact is secondary to the imminent threat that was controlled in August of 2003, after affecting 8.422 individuals, but before which we should be remain alert, since it is clear that the end of the SARS is not yet here; and that the appearance of a single case can mean the beginning of a new wave, joining the outbreak of other diseases such as the avian flu. In this regard, it is very important to know the epidemic bases of morbid-mortality of SARS and to keep a world wide surveillance in order to detect possible further cases.


title: Identification and characterization of a coronavirus packaging signal.

text: Previously, a mouse hepatitis virus (MHV) genomic sequence necessary for defective interfering (DI) RNA packaging into MHV particles (packaging signal) was mapped to within a region of 1,480 nucleotides in the MHV polymerase gene by comparison of two DI RNAs. One of these, DIssF, is 3.6 kb in size and exhibits efficient packaging, whereas the other, DIssE, which is 2.3 kb, does not. For more precise mapping, a series of mutant DIssF RNAs with deletions within this 1,480-nucleotide region were constructed. After transfection of in vitro-synthesized mutant DI RNA in MHV-infected cells, the virus product was passaged several times. The efficiency of DI RNA packaging into MHV virions was then estimated by viral homologous interference activity and by analysis of intracellular virus-specific RNAs and virion RNA. The results indicated that an area of 190 nucleotides was necessary for packaging. A computer-generated secondary structural analysis of the A59 and JHM strains of MHV demonstrated that within this 190-nucleotide region a stable stem-loop of 69 nucleotides was common between the two viruses. A DIssE-derived DI DNA which had these 69 nucleotides inserted into the DIssE sequence demonstrated efficient DI RNA packaging. Site-directed mutagenic analysis showed that of these 69 nucleotides, the minimum sequence of the packaging signal was 61 nucleotides and that destruction of the secondary structure abolished packaging ability. These studies demonstrated that an MHV packaging signal was present within the 61 nucleotides, which are located on MHV genomic RNA 1,381 to 1,441 nucleotides upstream of the 3' end of gene 1.


title: Sars.

text: NURSES MUST have the authority to order immediate infection prevention measures if they suspect a patient has SARS or any other dangerous infectious disease, the World Health Organization (WHO) has warned. Experts say the rapid spread of SARS in March proved there is no room for complacency in the management of patients with possible infections. The cities affected had exceptionally high standards of healthcare facilities.


title: History and recent advances in coronavirus discovery.

text: Human coronaviruses, first characterized in the 1960s, are responsible for a substantial proportion of upper respiratory tract infections in children. Since 2003, at least 5 new human coronaviruses have been identified, including the severe acute respiratory syndrome coronavirus, which caused significant morbidity and mortality. NL63, representing a group of newly identified group I coronaviruses that includes NL and the New Haven coronavirus, has been identified worldwide. These viruses are associated with both upper and lower respiratory tract disease and are likely common human pathogens. The global distribution of a newly identified group II coronavirus, HKU1, has not yet been established. Coronavirology has advanced significantly in the past few years. The SARS epidemic put the animal coronaviruses in the spotlight. The background and history relative to this important and expanding research area are reviewed here.


title: Kawasaki syndrome in Denmark.

text: OBJECTIVE To describe the epidemiologic characteristics of Kawasaki syndrome (KS) and to estimate national KS incidence rates among children in Denmark. METHODS A retrospective population-based study using hospital discharge records with a KS diagnosis for children younger than 15 years selected from the Danish National Hospital Register for 1981-2004. Incidence rates were calculated using the number of KS patients and corresponding census data. RESULTS During 1981-2004, 360 children younger than 15 years were hospitalized with KS in Denmark, with 73% younger than 5 years. In this age group, the average annual incidence of KS gradually increased from 1981 to 1999 and thereafter stabilized at 4.5 to 5.0 per 100,000 person-years. The incidence was greater for boys than for girls (RR = 1.6, 95% CI = 1.2-2.0) and was highest among infants younger than 1 year (4.5), declining with increasing age (P = 0.03). However, the age-specific decline in incidence was only observed for boys, whereas the incidence for girls remained unchanged by age. The median length of hospital stay was 12 days, and the incidence peaked in the winter months. CONCLUSIONS Major epidemiologic characteristics identified among Danish childhood KS are consistent with those described in previous studies, such as highest incidence among young children and winter-seasonality. The KS incidence rate among children younger than 5 years in Denmark increased steadily during the early study period (coinciding with global recognition of KS) and seems to have stabilized from 1998-1999 onwards. Although the incidence among Danish children was lower than that reported for several other European countries, differences in methodology challenge definite comparisons.


Veterinary public health.


title: Severe acute respiratory syndrome: quantitative assessment from chest radiographs with clinical and prognostic correlation.

text: OBJECTIVE This study examined findings of severe acute respiratory syndrome (SARS) on chest radiographs and presented a classification scheme using quantitative radiographic data supported by clinical parameters. MATERIALS AND METHODS Three radiologists who were blinded to the identity, diagnosis, treatment protocol, and outcome of each patient independently evaluated serial chest radiographs from 67 patients with confirmed SARS. In addition to the chest radiographic abnormalities and percentage of involvement, several quantitative improvement parameters, including the peak to 50% improvement time (PIT(50)), were collected. Correlation between PIT(50) and clinical parameters (duration of fever, cough, dyspnea, oxygen supplementation, intubation, and death) were evaluated using Wilcoxon's rank sum testing and Spearman's correlation. RESULTS The most common initial findings were unifocal air-space disease in the periphery of the lower lungs occurring a mean of 3.6 +/-2.4 (SD) days from symptoms onset. Peak abnormalities were seen at 10.4 +/- 2.9 days. PIT(50) was dependent on disease severity, showing a strong linear correlation with the clinical parameter duration of oxygen supplementation (r = 0.44, p = 0.0015). Three patterns of disease were recognized: pattern A (severe, 29.9%) with PIT(50) of more than 10 days, pattern B (typical, 44.8%) with PIT(50) of 10 or fewer days, and pattern C (mild, 25.4%) with minimal findings throughout the course of the disease. This classification was supported by collaborative clinical parameters. CONCLUSION The quantitative radiographic parameter PIT(50) has strong clinical correlation and can be used to differentiate severity of disease into severe, typical, and mild types.


title: Specific asparagine-linked glycosylation sites are critical for DC-SIGN- and L-SIGN-mediated severe acute respiratory syndrome coronavirus entry.

text: Severe acute respiratory syndrome (SARS) is caused by a newly emerged coronavirus (CoV) designated SARS-CoV. The virus utilizes angiotensin-converting enzyme 2 (ACE2) as the primary receptor. Although the idea is less clear and somewhat controversial, SARS-CoV is thought to use C-type lectins DC-SIGN and/or L-SIGN (collectively referred to as DC/L-SIGN) as alternative receptors or as enhancer factors that facilitate ACE2-mediated virus infection. In this study, the function of DC/L-SIGN in SARS-CoV infection was examined in detail. The results of our study clearly demonstrate that both proteins serve as receptors independently of ACE2 and that there is a minimal level of synergy between DC/L-SIGN and ACE2. As expected, glycans on spike (S) glycoprotein are important for DC/L-SIGN-mediated virus infection. Site-directed mutagenesis analyses have identified seven glycosylation sites on the S protein critical for DC/L-SIGN-mediated virus entry. They include asparagine residues at amino acid positions 109, 118, 119, 158, 227, 589, and 699, which are distinct from residues of the ACE2-binding domain (amino acids 318 to 510). Amino acid sequence analyses of S proteins encoded by viruses isolated from animals and humans suggest that glycosylation sites N227 and N699 have facilitated zoonotic transmission.


title: Coronavirus as agent of neonatal calf diarrhea in southern Chile.

text: Neonatal calf diarrhea is one of the most serious problems of cattle industry all over the world. Although the aetiology is complex, it is possible to assess that viruses play a very important role. During an investigation to study the importance of Rotavirus in enteric problems in calves and piglets, it was possible to demonstrate the presence, for the first time in southern Chile, of Coronavirus-like particles in a faecal sample of a 15 day old calf by means of electron microscopy. The importance of this diagnosis and some clinical characteristics of the disease are briefly discussed. Finally, the need for more research on this complex subject is recognized.


Inefficient infection of soluble receptor-resistant mutants of murine coronavirus in cells expressing MHVR2 receptor.


title: In situ hybridization for the detection and localization of porcine epidemic diarrhea virus in the intestinal tissues from naturally infected piglets.

text: Detection and localization of porcine epidemic diarrhea virus (PEDV) was studied by in situ hybridization with a nonradioactive digoxigenin-labeled probe in formalin-fixed, paraffin-embedded tissues from 10 naturally infected piglets. A 377-base pair cDNA probe for viral RNA encoding the membrane proteins of PEDV cell-culture-adapted strain V215/78 was generated by the reverse transcription polymerase chain reaction. In the retrospective study of pigs from herds with diarrhea, the 10 piglets naturally infected with PEDV had positive signals for PEDV by in situ hybridization. When intestinal tissues were hybridized with the PEDV probe, a strong signal was seen in the villus enterocytes of jejunum and ileum but not in the cecum and colon. Positive cells typically had dark brown reaction products in the cytoplasm. Scattered epithelial cells along the ileal Peyer's patches dome areas contained viral RNA. In one piglet, hybridization signal was also found in the duodenum. PEDV was not demonstrated in tissues outside of the intestinal tract. These findings indicate that jejunal and ileal villus enterocytes are the main target of PEDV replication during epizootic outbreaks of the disease.


title: Advances in the laboratory diagnosis of viral respiratory disease.

text: BACKGROUND In recent years significant advances in the laboratory diagnostics available to detect respiratory viral infections have been achieved. METHODS This article presents the types of diagnostic methods currently available to the practitioner, as well as those on the horizon. The article covers tissue culture, serology and direct examination as well as some rapid diagnostic techniques and molecular assays, previewing developing new technology. CONCLUSIONS Laboratory diagnosis will likely be clinically useful in some but not all cases of viral infection. As new diagnostic methods become widely available, it is increasingly important to develop guidelines for laboratory testing when viral illness is suspected.


title: A machine learning methodology for real-time forecasting of the 2019-2020 COVID-19 outbreak using Internet searches, news alerts, and estimates from mechanistic models.

text: We present a timely and novel methodology that combines disease estimates from mechanistic models with digital traces, via interpretable machine-learning methodologies, to reliably forecast COVID-19 activity in Chinese provinces in real-time. Specifically, our method is able to produce stable and accurate forecasts 2 days ahead of current time, and uses as inputs (a) official health reports from Chinese Center Disease for Control and Prevention (China CDC), (b) COVID-19-related internet search activity from Baidu, (c) news media activity reported by Media Cloud, and (d) daily forecasts of COVID-19 activity from GLEAM, an agent-based mechanistic model. Our machine-learning methodology uses a clustering technique that enables the exploitation of geo-spatial synchronicities of COVID-19 activity across Chinese provinces, and a data augmentation technique to deal with the small number of historical disease activity observations, characteristic of emerging outbreaks. Our model's predictive power outperforms a collection of baseline models in 27 out of the 32 Chinese provinces, and could be easily extended to other geographies currently affected by the COVID-19 outbreak to help decision makers.


title: [Emergency response plan for the neonatal intensive care unit during epidemic of 2019 novel coronavirus].

text: 2019 novel coronavirus (2019-nCoV) infection has been spreading in China since December 2019. Neonates are presumably the high-risk population susceptible to 2019-nCoV due to immature immune function. The neonatal intensive care unit (NICU) should be prepared for 2019-nCoV infections as far as possible. The emergency response plan enables the efficient response capability of NICU. During the epidemic of 2019-nCoV, the emergency response plan for the NICU should be based on the actual situation, including diagnosis, isolation, and treatment, as well as available equipment and staffing, and take into account the psychosocial needs of the families and neonatal care staff.


title: Cereulide and valinomycin, two important natural dodecadepsipeptides with ionophoretic activities.

text: Cereulide produced by Bacillus cereus sensu stricto and valinomycin synthesized mainly by Streptomyces spp. are natural dodecadepsipeptide ionophores that act as potassium transporters. Moreover, they comprise three repetitions of similar tetrapeptide motifs synthesized by nonribosomal peptide synthesis complexes. Resemblances in their structure find their reflections in the same way of action. The toxicity of valinomycin and cereulide is an effect of the disturbance of ionic equilibrium and transmembrane potential that may influence the whole organism and then cause fatal consequences. The vlm and ces operons encoding valinomycin and cereulide are both composed of two large, similar synthetase genes, one thioestrase gene and four other ORFs with unknown activities. In spite of the characterization of valinomycin and cereulide, genetic determinants encoding their biosynthesis have not yet been clarified.


title: Atypical and novel presentations of Coronavirus Disease 2019: a case series of three children.

text: Typical presentations of Coronavirus Disease 2019 (COVID19) including respiratory symptoms (cough, respiratory distress and hypoxia), fever and dyspnea are considered main symptoms in adults, but atypical presentation children could be a diagnostic challenge We report three children whose initial presentation was gastrointestinal, and in whom COVID-19 infection was found, concluding that cases of acute appendicitis, mesenteric adenitis and flank tenderness may mask and infection with this virus, which should therefore be investigated.


title: [Methods of treatment on different clinical stages in pancreatic cancer].

text: UNLABELLED Pancreatic cancer is a great surgical problem. Clinical observations confirm 10-15% level of R0 resection ability. Dissatisfying treatment effects results from: lach of simple screening tests, low specificity of methods used to diagnose less than 2 cm lesions, low availability of advanced diagnostic methods (endoscopic ultrasonography), anatomy of pancreatoduodenic area, ability of infiltration adhering structures and causing metastatic tumors. AIM Retrospective evaluation of treatment methods administered to patients with pancreatic cancer according to different clinical stages. MATERIAL AND METHODS Medical documentation of 97 patients treated during 2001-2006 period for pancreatic cancer was analyzed. The age of patients was 42-90 years (medium 66.04, SD 10.27). There were 47 women (48.45% of group) aged 43-90 (medium 67.89, SD 9.93) and 50 men (51.55%) aged 42-86 (medium 64.31, SD 10.64). RESULTS In analyzed group 87 tumors were located in the head of pancreas, 5 in the corpus and 5 in the cauda. In 13 cases (13.40%) partial resection of pancreas was performed. By passing anastomoses were applied in 45 cases: 26 (26.80% of total group) biliar and gastrointestinal, 19 (19.59%) only biliar anastomoses. Thoracovideoscopic splanchnicectomy was performed 26 times (26.80%) as a method of decreasing pain. 6 patients were treated with bile ducts drainage: endoscopic (4) or percutaneous (2). In 7 cases operations were limited to explorative laparotomy. Complication occurred in 5 (38.46%) cases following resection of the pancreas and 13 cases (15.38%) following paliative operations. Perioperative mortality accomplished 15.38% following pancreatic resection and 4.44% in remaining cases. CONCLUSIONS Pancreatic cancer is a neoplasm connected with unsuccessfull course. In most of cases diagnose is confirmed in clinical stage that disables resection with the aim of curation. Although huge progress in diagnostic methods, operational technique and perioperative care postoperative mortality is high. Videothoracoscopic splanchnicectomy is a worth reccomending method of decreasing pain.


title: Feline aminopeptidase N is a receptor for all group I coronaviruses.

text: Human coronavirus HCV-229E and porcine transmissible gastroenteritis virus (TGEV), both members of coronavirus group I, use aminopeptidase N (APN) as their cellular receptors. These viruses show marked species specificity in receptor utilization as they can only use APN of their respective species to initiate virus infection. Feline and canine coronaviruses are also group I coronaviruses. To determine whether feline APN could serve as a receptor for feline coronaviruses (FCoVs), we cloned the cDNA encoding feline APN (fAPN) by PCR from feline cells and stably expressed it in FCoV-resistant mouse or hamster cells. These became susceptible to infection with either of several biotypes of FCoVs. The expression of recombinant fAPN also made hamster and mouse cells susceptible to infection with other group I coronaviruses, including several canine coronavirus strains, transmissible gastroenteritis virus (TGEV), and human coronavirus HCV-229E. Thus, fAPN served as a functional receptor for each of these coronaviruses in group I. As expected, fAPN could not serve as a receptor for mouse hepatitis virus (MHV), a group II coronavirus which uses murine biliary glycoproteins as receptors. Thus, fAPN acts as a common receptor for coronaviruses in group I, in marked contrast to human and porcine APN glycoproteins which serve as receptors only for human and porcine coronaviruses, respectively. These observations suggest that cats could serve as a "mixing vessel" in which simultaneous infection with several group I coronaviruses could lead to recombination of viral genomes.


docid: nueoxytu

title: Trends in biotech literature 2005.


Influenza-like illness in residential care homes: a study of the incidence, aetiological agents, natural history, and health resource utilisation.


title: Feline aminopeptidase N serves as a receptor for feline, canine, porcine, and human coronaviruses in serogroup I.

text: Two members of coronavirus serogroup I, human respiratory coronavirus HCV-229E and porcine transmissible gastroenteritis virus (TGEV), use aminopeptidase N (APN) as their cellular receptors. These viruses show marked species specificity in receptor utilization, as HCV-229E can utilize human but not porcine APN, while TGEV can utilize porcine but not human APN. To determine whether feline APN could serve as a receptor for two feline coronaviruses in serogroup I, feline infectious peritonitis virus (FIPV) and feline enteric coronavirus (FeCV), we cloned the cDNA encoding feline APN (fAPN) by PCR from cDNA isolated from a feline cell line and stably expressed it in FIPV- and FeCV-resistant mouse and hamster cells. The predicted amino acid sequence of fAPN shows 78 and 77% identity with human and porcine APN, respectively. When inoculated with either of two biologically different strains of FIPV or with FeCV, fAPN-transfected mouse and hamster cells became infected and viral antigens developed in the cytoplasm. Infectious FIPV was released from hamster cells stably transfected with fAPN. The fAPN-transfected mouse and hamster cells were challenged with other coronaviruses in serogroup I including canine coronavirus, porcine coronavirus TGEV, and human coronavirus HCV-229E. In addition to serving as a receptor for the feline coronaviruses, fAPN also served as a functional receptor for each of these serogroup I coronaviruses as shown by development of viral antigens in the cytoplasm of infected mouse or hamster cells stably transfected with fAPN. In contrast, fAPN did not serve as a functional receptor for mouse hepatitis virus (MHV-A59), which is in serogroup II and utilizes mouse biliary glycoprotein receptors unrelated to APN. Thus, fAPN serves as a receptor for a much broader range of group I coronaviruses than human and porcine APNs. The human, porcine, and canine coronaviruses in serogroup I that are able to use fAPN as a receptor have previously been shown to infect cats without causing disease. Therefore, host factors in addition to receptor specificity apparently affect the virulence and transmissibility of nonfeline serogroup I coronaviruses in the cat.


title: Genetic analysis of determinants for spike glycoprotein assembly into murine coronavirus virions: distinct roles for charge-rich and cysteine-rich regions of the endodomain.

text: The coronavirus spike protein (S) forms the distinctive virion surface structures that are characteristic of this viral family, appearing in negatively stained electron microscopy as stems capped with spherical bulbs. These structures are essential for the initiation of infection through attachment of the virus to cellular receptors followed by fusion to host cell membranes. The S protein can also mediate the formation of syncytia in infected cells. The S protein is a type I transmembrane protein that is very large compared to other viral fusion proteins, and all except a short carboxy-terminal segment of the S molecule constitutes the ectodomain. For the prototype coronavirus mouse hepatitis virus (MHV), it has previously been established that S protein assembly into virions is specified by the carboxy-terminal segment, which comprises the transmembrane domain and the endodomain. We have genetically dissected these domains in the MHV S protein to localize the determinants of S incorporation into virions. Our results establish that assembly competence maps to the endodomain of S, which was shown to be sufficient to target a heterologous integral membrane protein for incorporation into MHV virions. In particular, mutational analysis indicated a major role for the charge-rich carboxy-terminal region of the endodomain. Additionally, we found that the adjacent cysteine-rich region of the endodomain is critical for fusion of infected cells, confirming results previously obtained with S protein expression systems.


title: Clinical issues and research in respiratory failure from severe acute respiratory syndrome.

text: The National Heart, Lung, and Blood Institute, along with the Centers for Disease Control and Prevention and the National Institute of Allergy and Infectious Diseases, convened a panel to develop recommendations for treatment, prevention, and research for respiratory failure from severe acute respiratory syndrome (SARS) and other newly emerging infections. The clinical and pathological features of acute lung injury (ALI) from SARS appear indistinguishable from ALI from other causes. The mainstay of treatments for ALI remains supportive. Patients with ALI from SARS who require mechanical ventilation should receive a lung protective, low tidal volume strategy. Adjuvant treatments recommended include prevention of venous thromboembolism, stress ulcer prophylaxis, and semirecumbent positioning during ventilation. Based on previous experience in Canada, infection control resources and protocols were recommended. Leadership structure, communication, training, and morale are an essential aspect of SARS management. A multicenter, placebo-controlled trial of corticosteroids for late SARS is justified because of widespread clinical use and uncertainties about relative risks and benefits. Studies of combined pathophysiologic endpoints were recommended, with mortality as a secondary endpoint. The group recommended preparation for studies, including protocols, ethical considerations, Web-based registries, and data entry systems.


title: Genetic interactions between an essential 3' cis-acting RNA pseudoknot, replicase gene products, and the extreme 3' end of the mouse coronavirus genome.

text: The upstream end of the 3' untranslated region (UTR) of the mouse hepatitis virus genome contains two essential and overlapping RNA secondary structures, a bulged stem-loop and a pseudoknot, which have been proposed to be elements of a molecular switch that is critical for viral RNA synthesis. It has previously been shown that a particular six-base insertion in loop 1 of the pseudoknot is extremely deleterious to the virus. We have now isolated multiple independent second-site revertants of the loop 1 insertion mutant, and we used reverse-genetics methods to confirm the identities of suppressor mutations that could compensate for the original insertion. The suppressors were localized to two separate regions of the genome. Members of one class of suppressor were mapped to the portions of gene 1 that encode nsp8 and nsp9, thereby providing the first evidence for specific interactions between coronavirus replicase gene products and a cis-acting genomic RNA element. The second class of suppressor was mapped to the extreme 3' end of the genome, a result which pointed to the existence of a direct base-pairing interaction between loop 1 of the pseudoknot and the genomic terminus. The latter finding was strongly supported by phylogenetic evidence and by the construction of a deletion mutant that reduced the 3' UTR to its minimal essential elements. Taken together, the interactions revealed by the two classes of suppressors suggest a model for the initiation of coronavirus negative-strand RNA synthesis.


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title: Structure and orientation of expressed bovine coronavirus hemagglutinin-esterase protein.

text: The sequence of the hemagglutinin-esterase (HE) gene for the Mebus strain of bovine coronavirus was obtained from cDNA clones, and its deduced product is a 47,700-kilodalton apoprotein of 424 amino acids. Expression of the HE protein in vitro in the presence of microsomes revealed N-terminal signal peptide cleavage and C-terminal anchorage but not disulfide-linked dimerization. Dimerization was observed only after expression in vivo, during which HE was also transported to the cell surface.


title: Preparing for the next influenza pandemic: a New South Wales perspective.

text: Pandemic influenza is one of a small number of infectious diseases that pose a significant global threat. Pandemic preparedness has accelerated around the world in recent years in response to the perceived increased risk of a pandemic developing following the emergence of H5N1 avian influenza in domestic poultry flocks in Asia, Africa and Europe. There is a hierarchy of pandemic plans - international, national, state, and local - and harmonisation of all of these is imperative for a coordinated and effective response. At the national and state levels, plans have been developed for a whole-of-government response to a pandemic, in addition to plans specifically for the health sector. It is inevitable that influenza pandemics will occur and careful planning is crucial to mitigate their potentially devastating effects.


title: Epidemiological and Clinical Aspects of COVID-19; a Narrative Review.

text: There are significant misconceptions and many obstacles in the way of illuminating the epidemiological and clinical aspects of COVID-19 as a new emerging epidemic. In addition, usefulness of some evidence published in the context of the recent epidemic for decision making in clinic as well as public health is questionable. However, misinterpreting or ignoring strong evidence in clinical practice and public health probably results in less effective and somehow more harmful decisions for individuals as well as subgroups in general populations of countries in the initial stages of this epidemic. Accordingly, our narrative review appraised epidemiological and clinical aspects of the disease including genetic diversity of coronavirus genus, mode of transmission, incubation period, infectivity, pathogenicity, virulence, immunogenicity, diagnosis, surveillance, clinical case management and also successful measures for preventing its spread in some communities.


title: Productivity losses associated with Fetal Alcohol Spectrum Disorder in New Zealand.

text: AIM To estimate the productivity losses due to morbidity and premature mortality of individuals with Fetal Alcohol Spectrum Disorder (FASD) in New Zealand (NZ). METHODS A demographic approach with a counterfactual scenario in which nobody in NZ is born with FASD was used. Estimates were calculated using (Census Year) 2013 data for the NZ population, the labour force, unemployment rate and average weekly wage, all of which were obtained from Statistics NZ. In order to estimate the number of FASD cases in 2013 and the related morbidity, the prevalence of FASD, obtained from the available epidemiological literature, was applied to the general population of NZ. Assumptions made on the level of impairment that would affect the ability of individuals with FASD to participate in the workforce or would reduce their productivity were based on data obtained from the current epidemiological literature. RESULTS In 2013, approximately 0.03% of the NZ workforce experienced a loss of productivity due to FASD-attributable morbidity and premature mortality, which translated to aggregate losses ranging from $NZ49 million to $NZ200 million - that is, 0.03% to 0.09% of the annual gross domestic product in NZ. These costs represent estimates for lost productivity attributable to FASD and do not include additional costs incurred by governmental and private entities including social costs, such as both higher costs and or less effective spending by the education, health and justice systems. CONCLUSION The estimated productivity losses associated with FASD further reinforces that effective FASD prevention as a primary public health strategy may be of significant value.


title: Spray cryotherapy is effective for bronchoscopic, endoscopic and open ablation of thoracic tissues.

text: OBJECTIVES Spray cryotherapy (SCT) delivers a liquid nitrogen spray via a catheter to produce cellular death. This study seeks to determine the histological changes after bronchoscopic, endoscopic and open SCT on tissues in the thoracic cavity. METHODS Yorkshire pigs underwent flexible bronchoscopy, endoscopy and thoracotomy for SCT of the airway, oesophagus and other intrathoracic structures, respectively. Variations in the duration and number of spray cycles for the same dosimetry were compared. RESULTS Bronchoscopic SCT of the airway resulted in cellular death up to the cartilage layer. Endoscopic SCT of the oesophagus led to cell death up to the adventitial layer. Tissue necrosis was severe in the lung, of full thickness in the pleura, but very superficial in the great vessels. The extracellular matrix (ECM) of treated tissues remained well-preserved. Having shorter but more cycles of SCT decreased the depth of the cellular necrosis. One pig developed ventricular fibrillation during the surgery and expired. CONCLUSIONS SCT causes reproducible tissue injury with the preserved ECM of most tissues within the thoracic cavity, making it enticing for ablation around vital structures like the great vessels with a decreased long-term risk. Further study is warranted to investigate the adverse events during SCT.


title: [Dynamics of peripheral blood B lymphocytes and natural killer cells in patients with severe acute respiratory syndrome].

text: OBJECTIVE To study the dynamics of peripheral blood B lymphocytes and natural killer (NK) cells in patients with severe acute respiratory syndrome (SARS). METHODS The absolute numbers of peripheral blood B lymphocytes and NK cells in 602 serial samples from 240 patients with SARS were counted, using flow cytometry, and compared with that of normal population. RESULTS The absolute numbers of peripheral blood B lymphocytes and NK cells in SARS patients were significantly lower than that of the normal population (P < 0.001) and were much lower in SARS patients with severe or extremely severe types, as compared with that of moderate or mild type cases (P < 0.001). The amount of B lymphocytes in recovery SARS patients increased at the 2nd week after onset, and gradually becoming normal at the 5th week of the disease onset. The number of NK cells was in the low level at onset, and keep decreasing at the 2nd week. However, it was increasing with the recovery of the disease, but did not reach to normal level at the 5th week after onset. CONCLUSION The absolute numbers of peripheral blood B lymphocytes and NK cells were associated with the severity of the disease, and detection of these two kinds of cells was useful for predicting the prognosis of SARS.


title: Activated surfaces for laser desorption mass spectrometry: application for peptide and protein analysis.

text: Thanks to the development of matrix assisted laser desorption/ionisation (MALDI), laser desorption based mass spectrometry became an essential method for the analysis of biomolecules. This review will discuss the various surface modifications used in combination with laser desorption mass spectrometry and their application for the analysis of peptides and proteins. In the first hand, some modified surfaces are designed to enhance the laser desorption/ionisation process; this includes the use of carbon, porous silicon surfaces and also immobilised matrix. In an other hand chemical and biochemical modified surfaces developed to isolate species with more or less specific interactions can be used for on-slide sample clean-up before MALDI-MS analysis. In addition, different experimental devices as mass spectrometers and microfluidic devices used for such a purpose will be presented.


title: Interference with cell cycle progression by parasitic genetic elements: sleeping beauty joins the club.

text: Transposable elements are discrete segments of DNA that have the distinctive ability to move and replicate within genomes. Similar to viruses, transposons are best viewed as molecular parasites that propagate themselves using resources of the host cell. Many viruses have developed strategies to modulate the host cell cycle machinery and cellular self-destruct mechanisms to maximize the chance for successful infection and the production of virus progeny. Recent evidence shows that transposable elements have also evolved mechanisms to modulate cell cycle progression for their own benefit. Thus, interference with the cell cycle seems to be a shared strategy of parasitic selfish genetic elements.


title: Down-regulation of TLR2, 3, 9 and Signaling Mediators, MyD88 and TRIF, Gene Transcript Levels in Patients with Kawasaki Disease Treated with IVIG.

text: Kawasaki disease (KD) is an acute febrile systemic vasculitis of childhood characterized by elevated levels of inflammatory mediators at the acute stage. High-dose intravenous immunoglobulin (IVIG) is well accepted as a conventional therapy for KD. The aim of the present study was to determine the expression level of Toll like receptors (TLRs) and their corresponding signaling mediators in PBMCs of IVIG-treated KD patients. TLR2, 3, 9 and signaling mediators, MyD88 and TRIF transcript levels were determined in PBMCs from 31 KD patients, before (acute phase), 2 weeks later (sub-acute phase) and 6 weeks later (convalescent phase) of IVIG therapy using real time PCR. The mean age of the patients was 3.6 years and 65% of subjects were male and 35% were female. 20 age-matched irrelevant febrile patients and 20 healthy subjects were included as control groups. Elevated levels of TLR2, MyD88, and TRIF gene transcripts were observed in the PBMCs at acute phase of untreated KD patients in compression with normal subjects. IVIG therapy resulted in significant decrease in TLR2, 3 and 9 (60-90%) as well as MyD88 and TRIF (60-70%) transcripts following 2 and 6 weeks. With Regard to significant up-regulation of MyD88 and TRIF at the acute phase of KD, our findings suggest TLR signaling pathway potential in KD pathogenesis and may also support the assumption of an infectious background in KD. Down-regulation of TLR members and corresponding mediators in IVIG treated patient suggest general TLR pathway suppression as a novel anti-inflammatory mechanism of IVIG.


Laboratory Diagnosis of COVID 19 - Perspectives.


Standing orders.


title: Disorders induced by the experimental infection of pigs with the porcine respiratory coronavirus (P.R.C.V.).

text: Five colostrum deprived hysterectomy-derived piglets were inoculated by the intratracheal route with a Porcine Respiratory Coronavirus (PRCV). The clinical effects and growth performances were compared with those in 5 similar piglets which were inoculated with Minimum Eagle Medium only. A transient hyperthermia, mild to severe dyspnoea, polypnoea and an obvious cessation of growth were recorded on infected pigs only whereas the control group did not show any adverse reaction after inoculation. The reasons for the variability of effects of experimental infection observed in previous trials, are discussed.


Suboptimal reporting of notifiable diseases in Canadian emergency departments: a survey of emergency physician knowledge, practices, and perceived barriers.


title: ACE2 receptor expression and severe acute respiratory syndrome coronavirus infection depend on differentiation of human airway epithelia.

text: Studies of patients with severe acute respiratory syndrome (SARS) demonstrate that the respiratory tract is a major site of SARS-coronavirus (CoV) infection and disease morbidity. We studied host-pathogen interactions using native lung tissue and a model of well-differentiated cultures of primary human airway epithelia. Angiotensin converting enzyme 2 (ACE2), the receptor for both the SARS-CoV and the related human respiratory coronavirus NL63, was expressed in human airway epithelia as well as lung parenchyma. As assessed by immunofluorescence staining and membrane biotinylation, ACE2 protein was more abundantly expressed on the apical than the basolateral surface of polarized airway epithelia. Interestingly, ACE2 expression positively correlated with the differentiation state of epithelia. Undifferentiated cells expressing little ACE2 were poorly infected with SARS-CoV, while well-differentiated cells expressing more ACE2 were readily infected. Expression of ACE2 in poorly differentiated epithelia facilitated SARS spike (S) protein-pseudotyped virus entry. Consistent with the expression pattern of ACE2, the entry of SARS-CoV or a lentivirus pseudotyped with SARS-CoV S protein in differentiated epithelia was more efficient when applied to the apical surface. Furthermore, SARS-CoV replicated in polarized epithelia and preferentially exited via the apical surface. The results indicate that infection of human airway epithelia by SARS coronavirus correlates with the state of cell differentiation and ACE2 expression and localization. These findings have implications for understanding disease pathogenesis associated with SARS-CoV and NL63 infections.


title: [Japanese college students' pessimism, coping strategies and anxiety: validation of the Japanese Defensive Pessimism Inventory (JDPI)].

text: The purpose of this study was to develop the Japanese Defensive Pessimism Inventory (JDPI), which measures defensive pessimism in an academic achievement situation for Japanese undergraduate students and differentiates between those who are realistically pessimistic and those who are defensively pessimistic. In Study 1,695 undergraduates completed the JDPI. A factor analysis revealed that the 24 items of the JDPI comprised four factors: Pessimism, Past experience, Positive reflectivity, and Effort. In Study 2, 618 undergraduates completed the JDPI, the Test Coping Strategy Scale, and the State-Trait Anxiety Inventory. The JDPI had high internal consistency and test-retest reliability. Defensive pessimists and strategic optimists had higher scores on the active coping strategy and lower scores on the avoidant-thinking coping strategy than did realistic pessimists. Furthermore, defensive pessimists and realistic pessimists had higher scores on the state anxiety and lower scores on the optimistic-thinking coping strategy than did strategic optimists. The results indicate that the JDPI had high concurrent validity.


title: Feline coronavirus participation in diarrhea of cats.

text: Fecal samples were examined for viruses participated in gastrointestinal disorders of cats, especially focusing on feline coronavirus (FCoV) by a reverse transcriptase-polymerase chain reaction assay. It was found that a primary viral pathogen was feline panleukopenia parvovirus (FPLV; 28.5% of the positive rate) and the secondary was FCoV (10.7%). Commonly reported clinical signs of cats of which feces were FCoV-positive were vomiting, diarrhea and dehydration with an exception of one serious case with concurrent FPLV infection.


title: Identification of angiotensin converting enzyme 2 in the rodent retina.

text: PURPOSE An active renin-angiotensin system has been found in the retina of rats and humans. Angiotensin-converting enzyme 2 (ACE2) is a recently discovered enzymatic homologue of Angiotensin-converting enzyme (ACE) that may be an important new component of the renin-angiotensin system (RAS). This study assesses the involvement of ACE2 in the normal and diabetic rodent retina and its modulation by ACE inhibition. METHODS Sprague-Dawley rats were randomised into three groups, control, diabetes, and diabetes plus ramipril, with diabetes induced with the cell toxin streptozocin and the study run for 24 weeks. ACE2 and ACE gene levels were measured using quantitative real-time polymerase chain reaction (QRT-PCR), ACE2 protein expression was confirmed by Western blotting, and ACE and ACE2 catalytic activity were measured using specific activity assays in the rat retina. Localisation of ACE2 mRNA and protein were determined by in situ hybridisation and immunohistochemistry, respectively. RESULTS ACE mRNA levels were decreased to approximately 50% in the diabetic retina, but ACE2 mRNA levels were not significantly changed. ACE but not ACE2 gene expression was influenced by ramipril treatment. Following immunostaining, both ACE2 and ACE protein were localised predominantly to the inner nuclear layer (INL) but also to photoreceptors. In the diabetic retina, ACE enzyme activity was decreased, whereas ACE2 enzyme activity was increased. CONCLUSIONS This study has identified ACE2 gene and catalytically active protein in the rodent retina. In diabetes, the major changes were a decrease in ACE but an increase in ACE2 enzymatic activity. The ACE inhibitor ramipril did not reduce ACE2 enzymatic activity.