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.


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.


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.


title: An update on C-reactive protein for intensivists.

text: This review aims to summarise the physiology of C-reactive protein (CRP), its possible roles and limitations as an inflammatory and infective marker in intensive care medicine, and also the emerging roles of CRP in the pathogenesis of cardiovascular and autoimmune diseases. Observational and animal studies on uses of CRP were retrieved from the PubMed database without any language restrictions. Quantitative data were not pooled because of the heterogeneity of patient characteristics and disparate ways in which CRP was studied. Serum CRP concentrations are determined by the synthetic rate of its production in the liver regulated predominantly by interleukin-6. It has a half-life of 19 hours and is relatively slow in its onset and offset in response to an acute inflammatory process when compared to procalcitonin. It has some favourable properties and limitations as an inflammatory marker An elevated CRP concentration is not specific to infections and the absolute CRP concentrations cannot be used to differentiate between bacterial, fungal and severe viral infections. The dynamic response of CRP to therapy that aims to modify the underlying inflammatory process and the clinical context of a patient are of pivotal importance when CRP concentrations are interpreted. CRP is found to be a significant partaker and prognostic factor in a wide range of cardiovascular and chronic diseases. In summary, CRP concentration is an important prognostic factor of many acute and chronic diseases. Serial CRP measurements may be useful to reflect a patient's response to therapy that aims to modify the underlying inflammatory process.


title: Study on the efficacies of splenic pedicle transection by using manual manipulation and Endo-GIA procedure for laparoscopic splenectomy.

text: Laparoscopy splenectomy (LS) was adopted in surgery from 1980s, it has become the main way of exploring for treating spleen diseases. Compared with conventional open surgery, LS has been gradually accepted by physicians and patients due to its advantages, including minimal surgical injury, less intraoperative blood loss, quick postoperative recovery, shorter hospital period, better cosmetic result, less risk of postoperative infections and improved postoperative quality of life Here, we try to investigate the splenic pedicle transection by using Endo-GIA (a linear stapling device) procedure and manual manipulation of secondary splenic pedicle for LS. A retrospective study was conducted on 60 patients who underwent LS. And patients were divided into two groups. 30 patients (group A) received splenic pedicle transection with Endo-GIA procedure and in the other 30 patients (group B) underwent secondary splenic pedicle transection for LS. Perioperative outcome measures of each group were recorded, including operation duration, intraoperative blood loss, postoperative flatus pass time, postoperative complications, drainage duration, hospital cost and length of hospital stay. Surgeries were successfully achieved in 60 patients. The operative duration of group A was significantly shorter than that of group B. However, group B was significantly superior over Endo-GIA group in terms of the intraoperative blood loss, postoperative flatus pass time, drainage duration, length of hospital stay and total cost of hospital stays. No significant differences were observed in postoperative fever, ascites and hyperamylasemia between two groups. Both of these two approaches for LS are safe and feasible. However, compared with Endo-GIA procedure, manual manipulation of secondary splenic pedicle for LS may leading to less intraoperative blood loss, results in less hospital expense, and hence can be widely adopted in clinical practice.


Severe acute respiratory syndrome (SARS) in Hong Kong's ICUs: the lessons learned.


title: Simulation and visualization of flow pattern in microarrays for liquid phase oligonucleotide and peptide synthesis.

text: Microfluidic microarrays have been developed for economical and rapid parallel synthesis of oligonucleotide and peptide libraries. For a synthesis system to be reproducible and uniform, it is crucial to have a uniform reagent delivery throughout the system. Computational fluid dynamics (CFD) is used to model and simulate the microfluidic microarrays to study geometrical effects on flow patterns. By proper design geometry, flow uniformity could be obtained in every microreactor in the microarrays.


title: [Viral infection].

text: Viral infections, particularly in the fetus or newborn and in immunocompromised individuals, can produce severe, even fatal disease. Indeed Herpesviridae infections are the major cause of morbidity and mortality in them. Antiviral drugs have been used for prophylaxis and treatment for some viral infections. An alternative approach is to boost the host immune defences. Administration of pooled human sera have been used for prophylaxis or alleviation of clinical severity of ongoing infections with measles, CMV, VZV and hepatitis A and B and so on. Human monoclonal antibody preparations, which have now being developed, may overcome the problems inherent in the pooled sera. These include high volume load, difficulty of quality control and adventitious transmission of unknown pathogenic agents.


title: Planning for Epidemics and Pandemics: Assessing the Potential Impact of Extended Use and Reuse Strategies on Respirator Usage Rates to Support Supply-and-Demand Planning Efforts.

text: During epidemics and pandemics healthcare personnel (HCP) are on the front line of disease containment and mitigation. Personal protective equipment (PPE), such as NIOSH-approved N95 filtering facepiece respirators (FFRs), serve an important role in minimizing HCP risks and are in high demand during public health emergencies. Because PPE demand can exceed supply, various public health strategies have been developed to reduce the rate of PPE consumption as supply dwindles. Extended use and limited reuse of N95 FFRs are strategies advocated by many governmental agencies used to increase the number of times a device can be used. Increased use of respirators designed for reuse-such as powered air-purifying respirators (PAPRs) and elastomeric half-mask and full facepiece air-purifying respirators- is another option designed to reduce the continuous need for new devices as the daily need for respirator use increases. Together, these strategies are designed to reduce the number of PPE units that must be discarded daily and, therefore, extend the longevity of available supply. The purpose of this paper is to theoretically estimate the impact of extended use and limited reuse strategies for N95 FFRs and the increased use of reusable respirator options on PPE consumed. The results suggest that a considerable reduction in PPE consumption would result from extended use and limited reuse of N95 FFRs and the increased use of respirators designed for reuse; however, the practical benefits must be balanced with the risks and economic costs. In addition, extended use and reuse strategies must be accompanied by proper procedures to reduce risk. The study is designed to support epidemic and pandemic PPE supply and demand planning efforts.


title: cis-preferential stimulation of alfalfa mosaic virus RNA 3 accumulation by the viral coat protein.

text: RNA 3 of alfalfa mosaic virus (AIMV) encodes the movement protein P3 and the viral coat protein (CP) which is translated from the subgenomic RNA 4. RNA 3 is able to replicate in tobacco plants transformed with the AIMV replicase genes P1 and P2 (P12 plants). Frameshifts or deletions in the P3 gene have little effect on RNA 3 accumulation in P12 protoplasts whereas such mutations in the CP gene result in a 100-fold reduction of plus-strand RNA 3 accumulation. When P12 protoplasts were inoculated with a mixture of a RNA 3 mutant with a deletion in the P3 gene and a mutant with a deletion in the CP gene, CP expressed by the P3 mutant was unable to upregulate plus-strand RNA accumulation of the CP mutant. However, when a wild-type CP gene and subgenomic promoter were inserted in a RNA 3 mutant with a defective CP gene, the mutant accumulated at wild-type levels. It is concluded that the function of CP in plus-strand RNA 3 accumulation acts in cis and cannot be complemented in trans. In P12 plants, P3 and CP mutants were able to complement each other at low and variable levels. This complementation in plants appeared to be correlated with the occurrence of recombination to wild-type RNA 3.


title: [Microwave ablation of liver tumors as a new instrument for minimally invasive liver surgery].

text: Incidence of primary and secondary liver tumors is increasing. Hepatic resection remains the treatment of choice for hepatic tumors. For various reasons the vast majority of patients with liver tumors are not suitable for resection. These patients are candidates for several image-guided focal thermal ablative therapies as alternatives to resection. Currently available ablative techniques include cryotherapy, radiofrequency ablation (RFA), microwave ablation (MWA), laser ablation, high-intensity focused ultrasound ablation (HIFU), and ethanol injection. Presently RFA is most widely heat-based technology used for treatment of liver malignancies due to its availability, efficacy and low complication rates. However, RFA can be time-consuming and associated with higher recurrence rates in larger lesions. MWA is a new thermal ablative technique that uses electromagnetic energy to produce coagulation necrosis. MWA has several advantages over RFA such as an improved convection profile, consistently higher intratumoral temperatures, larger ablation volumes, faster ablation times, and the option of using multiple antennae simultaneously. We report our first experience using MWA and a Coviden Valleyab 915 MHz generator for ablation of liver tumor with respect to our previous experience with RFA. Further this study reviews current literature on the RFA and MWA for the treatment of the liver malignances. In our opinion although experience is limited MWA appears to be a safe and effective technology for hepatic tumor ablation and in some cases may be superior alternative to RFA.


title: Expanding ICU facilities in an epidemic: recommendations based on experience from the SARS epidemic in Hong Kong and Singapore.

text: Epidemics have the potential to severely strain intensive care resources and may require an increase in intensive care capability. Few intensivists have direct experience of rapidly expanding intensive care services in response to an epidemic. This contribution presents the recommendations of an expert group from Hong Kong and Singapore who had direct experience of expanding intensive care services in response to the epidemic of severe acute respiratory syndrome. These recommendations cover training, infection control, staffing, communication and ethical issues. The issue of what equipment to purchase is not addressed. Early preparations should include fit testing of negative pressure respirators, training of reserve staff, sourcing of material for physical modifications to the ICU, development of infection control policies and training programmes, and discussion of triage and quarantine issues.


Classification of Diseases and Injuries.


title: The Coronaviridae now comprises two genera, coronavirus and torovirus: report of the Coronaviridae Study Group.

text: At the April 1992, mid-term meeting of the International Committee on Taxonomy of Viruses (ICTV) a proposal from the Coronaviridae Study Group (CSG) to include the torovirus genus in the Coronaviridae was accepted. Following another proposal, the arterivirus genus was removed from the Togaviridae but not assigned to another family. The arteriviruses have some features in common with the Coronaviridae but also have major differences. After much debate, culminating in September 1992, it was decided that the CSG would not recommend inclusion of arterivirus in the Coronaviridae. It was agreed that (a) the nomenclature used for coronavirus genes, mRNAs and polypeptides (Cavanagh et al., 1990) should be used for toroviruses, (b) that the small (about 100 amino acids) membrane-associated protein, which is distinct from the integral membrane glycoprotein M, associated with virions of infectious bronchitis (Liu & Inglis, 1991) and transmissible gastroenteritis (Godet et al., 1992) coronaviruses would be referred to by the acronym sM (lower case 's') and (c) that 'pol' (polymerase) should be used as a working term for gene 1, which comprises open reading frames (ORFs) 1a and 1b in both genera of the Coronaviridae.


title: Relationship between ambient air pollution and daily mortality of SARS in Beijing.

text: OBJECTIVE To study the relationship between ambient air pollution and daily mortality of SARS in Beijing. METHODS The approach of time-series Poisson regression was used to assess the relationship between daily SARS mortality, ambient air pollution, and other factors from April 25 to May 31, 2003 in Beijing. RESULTS An increase of each 10 microg/m3 over a 5-day moving average of PM10, SO2 and NO2 corresponded to 1.06 (1.00-1.12), 0.74 (0.48-1.13) and 1.22 (1.01-1.48) relative risks (RRs) of daily SARS mortality, respectively. The relative risks (RRs) values depended largely on the selection of lag days. CONCLUSION The daily mortality of SARS might be associated with certain air pollutants in Beijing.


title: Clinical application of long-term Palindrome catheter in hemodialysis patients.

text: INTRODUCTION This study was conducted to compare catheter function, dialysis adequacy, and dialysis-related complications among patients receiving long-term dialysis via Palindrome catheter, Permcath catheter, and arteriovenous fistula (AVF) as vascular access. MATERIALS AND METHODS Forty-three patients undergoing dialysis with Permcath catheter, 49 with Palindrome catheter, and 56 with AVF were recruited. Urea clearance (KT/V), urea reduction rate, and the highest blood flow at the arteriovenous junction during dialysis were determined during the dialysis sessions. Catheter-related infection, catheter-associated thrombosis, and annual patency rate were also evaluated. RESULTS In patients using Permcath catheter for dialysis, the incidence of secondary renal injury (metabolic diseases, hypertension, and ischemic kidney diseases) was 73.1%, which was significantly higher than that in patients with AVF (51.5%; P = .001). In the Palindrome group, maximum blood flow, KT/V, urea reduction rate, and annual patency rate were significantly higher than those in the Permcath group, and the incidence of access-related infection was significantly higher than that in the AVF group. In the Palindrome group, the prevalence of thromboembolism was 30.6%, which was significantly lower than that in the Permcath group (46.5%), but higher than that in the AVF group (5.4%). CONCLUSIONS For dialysis patients, Palindrome catheter was superior to Permcath catheter and comparable with the AVF in terms of the maximum blood flow, dialysis adequacy, and annual patency rate. Dialysis with Palindrome catheter has a high infection rate and a high incidence of thromboembolism as in the dialysis with Permcath catheter.


title: Human neutrophil elastase in RSV bronchiolitis.

text: Acute bronchiolitis is the most common lower respiratory tract infection in young children and may be life-threatening in those with underlying cardiac or respiratory conditions. We evaluated the nasal and serum levels of human neutrophil elastase (HNE) in patients with acute respiratory syncytial virus (RSV) bronchiolitis and investigated the correlation of these levels with illness severity. Fifty-one patients (28 boys, 23 girls) with acute bronchiolitis positive for RSV by direct immunoenzyme assay in nasal secretions (Group A) were studied. Thirty healthy children (17 boys, 13 girls) constituted the control group (Group B). Subjects in both groups were matched for age and gender. The ages (mean+/-SE) in Groups A and B were 4.5+/-0.41 and 5.0+/-0.65 mo, respectively. Venous blood and nasal secretions were taken from patients in group A on 1, 5, and 15 days after admission and once from controls (Group B) for determinations of HNE in nasal lavage and serum, as well as white blood counts (WBC). The peripheral blood eosinophil and neutrophil counts were elevated in 22/51 patients (43.1%) and 15/51 patients (29.4%), respectively. In nasal lavage specimens, neutrophils represented>or=75% and eosinophils>2% of all cells in 42/51 (82.0%) patients and 11/51 (21.5%) patients, respectively. There was strong correlation between the level of HNE and the percentage of neutrophils in nasal lavage (r=0.92). The mean nasal HNE concentrations of the patients on 1, 5, and 15 days after admission were higher than those of Group B (p<0.0001, p<0.001, p<0.001, respectively). Mean serum HNE concentrations on 1, 5, and 15 days after admission were higher in Group A than in Group B (p<0.0001, p<0.0001, p<0.0001, respectively). Nasal and serum HNE concentrations showed no correlations with the clinical score of disease severity (r=0.28 and r=0.29, respectively). This study shows that (a) serum and nasal HNE concentrations were significantly higher in RSV bronchiolitis patients than in controls, (b) they did not return to normal after the respiratory symptoms had improved, and (c) they showed no significant correlations with clinical score of severity. The results indicate that neutrophils contribute significantly to airway inflammation in these subjects and HNE levels in serum and nasal lavage may be useful markers of inflammation in acute RSV bronchiolitis.


title: Are we ready for pandemic influenza?

text: During the past year, the public has become keenly aware of the threat of emerging infectious diseases with the global spread of severe acute respiratory syndrome (SARS), the continuing threat of bioterrorism, the proliferation of West Nile virus, and the discovery of human cases of monkeypox in the United States. At the same time, an old foe has again raised its head, reminding us that our worst nightmare may not be a new one. In 2003, highly pathogenic strains of avian influenza virus, including the H5N1 and H7N7 subtypes, again crossed from birds to humans and caused fatal disease. Direct avian-to-human influenza transmission was unknown before 1997. Have we responded to these threats by better preparing for emerging disease agents, or are we continuing to act only as crises arise? Here we consider progress to date in preparedness for an influenza pandemic and review what remains to be done. We conclude by prioritizing the remaining needs and exploring the reasons for our current lack of preparedness for an influenza pandemic.


title: Ribonucleoprotein of avian infectious bronchitis virus.

text: The ribonucleoprotein (RNP) of avian infectious bronchitis virus (IBV) was examined by electron microscopy after shadowing with carbon/platinum. Linear RNP strands up to 6.7 microns in length, from three IVB strains, were sensitive to both pancreatic RNase and to proteases. These strands were obtained from spontaneously disrupted complete particles but not from disrupted incomplete particles that lacked RNP. They were also released from Nonidet P40-disrupted particles and could be isolated on sucrose density gradients at a density of 1.27 g/ml. In some cases, helical RNP complexes associated with virus particles were observed that were similar to RNPs of human coronavirus strain 229E and mouse hepatitis virus strain 3.


title: Pneumonia in pregnancy.

text: OBJECTIVE Historically, pneumonia during pregnancy has been associated with increased morbidity and mortality compared with nonpregnant women. The goal of this article is to review current literature describing pneumonia in pregnancy. This review will identify maternal risk factors, potential complications, and prenatal outcomes associated with pneumonia and describe the contemporary management of the varied causes of pneumonia in pregnancy. RESULTS Coexisting maternal disease, including asthma and anemia, increase the risk of contracting pneumonia in pregnancy. Neonatal effects of pneumonia in pregnancy include low birth weight and increased risk of preterm birth, and serious maternal complications include respiratory failure. Community-acquired pneumonia is the most common form of pneumonia in pregnancy, with Streptococcus pneumoniae, Haemophilus influenzae, and Mycoplasma pneumoniae accounting for most identified bacterial organisms. Beta-lactam and macrolide antibiotics are considered safe in pregnancy and are effective for most community-acquired pneumonia in pregnancy. Viral respiratory infections, including varicella, influenza, and severe acute respiratory syndrome, can be associated with maternal pneumonia. Current antiviral and respiratory therapies can reduce maternal morbidity and mortality from viral pneumonia. Influenza vaccination can reduce the prevalence of respiratory hospitalizations among pregnant women during influenza season. Pneumocystis pneumonia continues to carry significant maternal risk to an immunocompromised population. Prevention and treatment of Pneumocystis pneumonia with trimethoprim/sulfamethoxazole is effective in reducing this risk. CONCLUSIONS Prompt diagnosis and treatment with contemporary antimicrobial therapy and intensive care unit management of respiratory compromise has reduced the maternal morbidity and mortality due to pneumonia in pregnancy. Prevention with vaccination in at-risk populations may reduce the prevalence and severity of pneumonia in pregnant women.


title: CEACAM2 positively regulates integrin αIIbβ3-mediated platelet functions.

text: Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is an Ig-ITIM superfamily member that regulates integrin αIIbβ3 function. We hypothesized that its twin protein, CEACAM2, exerts a similar physiologic role in murine platelets. CEACAM2-deficient mice (Cc2-/-) displayed prolonged tail bleeding times and increased volume of blood loss. Cc2-/- platelets have moderate integrin αIIbβ3-mediated functional defects with impaired kinetics of platelet spreading on fibrinogen and type I collagen and delayed kinetics in the retraction of fibrin clots in vitro. This functional integrin αIIbβ3 defect could not be attributed to altered integrin αIIbβ3 expression. Cc2-/- platelets displayed normal 'inside-out' signaling properties as demonstrated by normal agonist-induced binding of soluble fluorescein isothiocyanate (FITC)-fibrinogen and JON/A antibody binding. This data provides direct evidence that disruption of CEACAM2 induces a moderate integrin αIIbβ3-mediated platelet function defect, and that CEACAM2 is essential to maintain a normal integrin αIIbβ3-mediated platelet function.


title: Leader switching occurs during the rescue of defective RNAs by heterologous strains of the coronavirus infectious bronchitis virus.

text: A defective RNA (D-RNA), CD-61, derived from the Beaudette strain of the avian coronavirus infectious bronchitis virus (IBV), was rescued (replicated and packaged) using four heterologous strains of IBV as helper virus. Sequence analysis of the genomic RNA from the four heterologous IBV strains (M41, H120, HV10 and D207) identified nucleotide differences of up to 17% within the leader sequence and up to 4.3% within the whole of the adjacent 5' untranslated region (UTR). Analysis of the 5' ends of the rescued D-RNAs showed that the Beaudette leader sequence, present on the initial CD-61, had been replaced with the corresponding leader sequence from the helper IBV strain but the adjacent 5' UTR sequence of the rescued D-RNAs corresponded to the original CD-61 Beaudette sequence. These results demonstrated that the phenomenon of leader switching previously identified for the coronaviruses murine hepatitis virus and bovine coronavirus (BCoV) also occurred during the replication of IBV D-RNAs. Three predicted stem-loop structures were identified within the 5' UTR of IBV. Stem-loop I showed a high degree of covariance amongst the IBV strains providing phylogenetic evidence that this structure exists and is potentially involved in replication, supporting previous observations that a BCoV stem-loop homologue was essential for replication of BCoV defective interfering RNAs.


title: Comparative study of the effect of tocotrienols and -tocopherol on fasting serum lipid profiles in patients with mild hypercholesterolaemia: a preliminary report.

text: OBJECTIVES Hypercholesterolaemia is a risk factor for cardiovascular diseases. Tocotrienols reportedly possess hypocholesterolaemic activity. This study examined the effect of tocotrienols (T3) in TOCOVIDTM Suprabio TM on serum lipids. Patients and Methods :A randomised (2:1), open-label study of patients with mild hypercholesterolaemia (= 5.18mmol/L to <7.77mmol/L) and one additional cardiovascular risk factor was carried out. Subjects received either tocotrienols (as TOCOVIDTM Suprabio TM ) (n=28) or vitamin E (a-tocopherol) 500mg daily (n=16). Fasting lipids were compared at baseline and after 4 weeks therapy. RESULTS Following 4 weeks therapy, mean +/- SD total cholesterol declined significantly in the tocotrienol group (from 6.10+/-0.66 to 5.47+/-1.16; P=0.02) compared to the a-tocopherol group (from 5.92+/-0.52 to 5.47+/-0.76; P>0.05). Mean LDL-C levels (mmol/L) were also significantly reduced in the tocotrienol group (3.82+/-0.85 to 3.24+/-1.26; P=0.04), but not in those on a-tocopherol (3.84+/-0.75 to 3.28+/-0.94; P>0.05). There were no significant changes in HDL-C and triglycerides in both groups. The tocotrienol group experienced a net decline in TG (7.1+/-31.4 %; P>0.05) while the a-tocopherol group had a net increase at week 4 (38.6+/-61.7%; P>0.05). CONCLUSION The study adds to existing evidence of the favourable effect of tocotrienols on total cholesterol and LDL-C. However, the results need further evaluation.


title: Pathogenesis of avian flu H5N1 and SARS.

text: Avian influenza A (H5N1) and severe acute respiratory syndrome (SARS) coronavirus are infections that cause a severe viral pneumonia leading to acute respiratory dysfunction syndrome and carry a high case-fatality rate. We have investigated innate immune responses to both viruses using primary human macrophages and respiratory epithelial cells as in vitro models. In contrast to human influenza A H1NI viruses, the H5N1 viruses hyper-induce cytokines (tumour necrosis factor [TNF]alpha, interferon beta) and chemokines (IP10, MIP1alpha, MCP) in in vitro cultures of primary human macrophages. A similar differential effect is observed in primary human bronchial epithelial cells and in type 2 pneumocytes although TNFalpha is not induced in respiratory epithelial cells. The cell signalling pathways responsible for this differential effect remain to be explored. Preliminary data suggest that such differential signalling involves p38 MAP kinase rather than NF-kappaB. SARS coronavirus infection of primary human macrophages is associated with a strong induction of chemokines without an associated type 1 interferon response. These observations may be relevant in disease pathogenesis.


title: [At least 68 vaccine candidates under development].

text: The development of vaccines against SARS-CoV-2 is progressing at an unparalleled speed. As of the 29th of March, there were at least 68 vaccine candidates comprising several different vaccine designs, including whole killed virus, subunit, attenuated, viral vector, DNA and mRNA vaccines. Whilst it usually takes 10-15 years to develop a vaccine, it has only taken just over 9 weeks from the publication of the viral genetic sequence for the first vaccine candidate to reach clinical testing. Development has been expediated by using existing technological platforms and by performing preclinical and clinical testing simultaneously.


title: Use of the trellis device in the management of deep vein thrombosis: a retrospective single-center experience.

text: OBJECTIVES The Trellis thrombectomy system (Covidien) is one of the newer devices that incorporates isolated pharmacomechanical thrombectomy and thrombolysis (PMT) for treatment of deep venous thrombosis (DVT). We conducted a retrospective review of patients with upper- and lower-extremity DVT managed with the Trellis thrombectomy system at our center. METHODS All patients with symptomatic DVT who presented to our center between April 2010 and April 2011 who underwent PMT by the Trellis device were included in this retrospective review. RESULTS Twenty-eight patients (mean age, 46.4 ± 21.2 years) presented with symptoms with a mean duration of 1.3 ± 1.8 months. Eighty-six percent had 100% occlusion on admission, while 14.3% had 70%-90% stenosis. The mean lytic dose used was tPA 20.7 ± 12 mg. The mean Trellis treatment time was 25.1 ± 11.5 minutes. Grade 3 lysis was achieved in 23 of 28 patients (85.8%), while grade 2 lysis was achieved in 14.2%. Mean total hospital stay was 2.6 ± 2.7 days. Postprocedure symptom resolution was 100%, and there was no reocclusion in 78.6% of patients at 1 year. At 12 months, the patency rate (primary or secondary) was 80% as determined by Doppler ultrasound. CONCLUSIONS In patients with DVT involving the ilio-femoral and the upper-extremity vessels, the use of the Trellis device was associated with a high technical success rate as well as a satisfactory 12-month patency rate. Moreover, this strategy was associated with reduced lytic dose, shorter treatment time and hospital stay, and no bleeding complications.