title: Coronavirus: how a large RNA viral genome is replicated and transcribed.

text: Coronaviruses are important human and animal pathogens and contain an extraordinarily long (27-31 kb) RNA genome. Its RNA synthesis involves complex mechanisms of regulation, similar to those of DNA viruses. In this treatise, mouse hepatitis virus (MHV) is used as a model for the discussion of the mechanism of viral RNA synthesis. We show that MHV RNA synthesis requires interactions of multiple RNA components, which are likely mediated by protein-RNA and protein-protein, as well as RNA-RNA, interactions. This virus also provides a unique example of a discontinuous transcription mechanism, which involves a trans-acting RNA component. Finally, study of the cis-acting signals for the various steps of RNA synthesis revealed an insight into the regulation of viral RNA synthesis. This discussion suggests that the regulation of RNA synthesis in coronavirus is more complex than previously thought possible for RNA viruses. Coronavirus RNA transcription and replication may serve as a paradigm of RNA synthesis for RNA viruses in general.


title: The provision of sophisticated critical care beyond the hospital: lessons from physiology and military experiences that apply to civil disaster medical response.

text: OBJECTIVE The provision of sophisticated medical care in an austere environment is challenging. During and after a mass casualty event, it is likely that critical care services will be needed beyond an intensive care unit (ICU) setting. The objective of this article is to explore existing ICU care systems such as military aeromedical transport that may be applicable to disaster medicine and to providing critical care outside of an ICU setting. RESULTS The U.S. Air Force Critical Care Aeromedical Transport (CCAT) Teams were developed in 1994 in response to an unmet military need for long-range air transport of critically ill and injured patients. This system has transported several thousand ICU patients and is an applicable model for the future development of extrahospital critical care capabilities needed during a disaster. We also discuss civilian aeromedical critical care systems, the types of medical devices used, and their applicability to disaster medical response. CONCLUSION The U.S. Air Force CCAT Team program, as well as many civilian critical care air ambulance services, provides a workable starting point for the development of disaster medical critical care response capabilities for disaster medical systems.


title: Tongue rotation for reconstruction after rostral hemiglossectomy for excision of a liposarcoma of the rostral quadrant of the tongue in a dog.

text: A 15-year-old female beagle dog was presented for a lingual liposarcoma. Full staging of the disease did not show any evidence of metastasis. A tongue rotation for reconstruction after rostral hemiglossectomy was performed after removal of the mass. This surgical technique may be useful in tongue reconstruction after trauma or tumor excision.


title: Influenza virus NS vectors expressing the mycobacterium tuberculosis ESAT-6 protein induce CD4+ Th1 immune response and protect animals against tuberculosis challenge.

text: Infection with Mycobacterium tuberculosis remains a major cause of morbidity and mortality all over the world. Since the effectiveness of the only available tuberculosis vaccine, Mycobacterium bovis bacillus Calmette-Guérin (BCG), is suboptimal, there is a strong demand to develop new tuberculosis vaccines. As tuberculosis is an airborne disease, the intranasal route of vaccination might be preferable. Live influenza virus vaccines might be considered as potential vectors for mucosal immunization against various viral or bacterial pathogens, including M. tuberculosis. We generated several subtypes of attenuated recombinant influenza A viruses expressing the 6-kDa early secretory antigenic target protein (ESAT-6) of M. tuberculosis from the NS1 reading frame. We were able to demonstrate the potency of influenza virus NS vectors to induce an M. tuberculosis-specific Th1 immune response in mice. Moreover, intranasal immunization of mice and guinea pigs with such vectors induced protection against mycobacterial challenge, similar to that induced by BCG vaccination.


title: Immunological characterization of the spike protein of the severe acute respiratory syndrome coronavirus.

text: Severe acute respiratory syndrome (SARS) is a novel infectious disease caused by the SARS-associated coronavirus (SARS-CoV). There are four major structural proteins in the SARS-CoV, including the nucleocapsid, spike, membrane, and small envelope proteins. In this study, two sets of truncated fragments of spike protein were generated, the first were approximately 210-bp nonoverlapping fragments and the second were overlapping segments of 750 to 900 bp. From these 23 fragments, we identified a fragment of 259 amino acids (amino acids 441 to 700) that is a major immunodominant epitope. This fragment was highly expressed, and the purified fragment C could detect all 33 SARS patient serum samples tested, collected from 7 to 60 days after the onset of fever, but had no reactivity with all 66 healthy human serum samples tested. Thus, fragment C of spike protein was identified as an immunodominant antigen and could be used for serological detection of SARS-CoV infection.


title: Coronavirus-like particles and other agents in the faeces of children in Efate, Vanuatu.

text: Between June 1979 and July 1980, faecal specimens were collected from 100 children in Efate, Vanuatu, and examined for the presence of virus-like particles by direct negative staining electron microscopy. Of the children, 40 were hospital patients 22 of whom had gastroenteritis and 18 had other illnesses. The remaining 60 children were apparently healthy controls. Coronavirus-like particles were the most common agents encountered and were detected in approximately 20% of all three groups of children. Particles with the morphological features of calicivirus, astrovirus, adenovirus and parvovirus were also detected.


title: Emerging treatment options for venous ulceration in today's wound care practice.

text: UNLABELLED Lower-extremity ulcers represent the largest group of ulcers presenting to an outpatient wound care clinic and, of those, ulcers due to venous insufficiency and venous hypertension make up the largest subgroup of these ulcers. Interventions for chronic venous ulcers have evolved to painless, minimally invasive, office-based procedures performed under local anesthesia. Recent advances in the endovascular management of lower-extremity superficial venous insufficiency have the potential to significantly enhance initial and long-term management of these patients, as minimally invasive procedures provide faster recoveries and fewer procedural risks. Early intervention for venous insufficiency has been shown to significantly decrease long-term ulcer recurrence rates, and may increase healing of venous ulcers as well. PURPOSE The purpose of this review and algorithm is to enhance understanding of venous ulceration and its underlying causes. Venous anatomy and pathophysiology will be reviewed. The etiology of chronic venous ulceration will be examined. Current practice guidelines and clinical outcomes will be reviewed. The newest treatment options, including minimally invasive therapy will be described. The current literature will be reviewed. A new algorithm for treatment that integrates the endovascular treatment of venous insufficiency into the current standard care for venous leg ulceration will be proposed. METHOD A literature review was performed to review all current treatments for venous ulceration. Treatments that have Level I evidence (with a grade of recommendation of A or B) to support their use for venous leg ulceration were selected for incorporation into a new treatment algorithm. The level-of-evidence and strength-of-recommendation scheme used in the algorithm is based upon the system used by the Wound Healing Society in its chronic wounds clinical practice guidelines. CONCLUSION It is intended that this new algorithm and approach to treatment will improve the immediate care of venous leg ulcer patients, reduce recurrence rates, increase patient satisfaction, and potentially expedite initial wound healing in the outpatient wound clinic setting.


title: Possible relationship between streptococcal pyrogenic exotoxin A and Kawasaki syndrome in patients older than six months of age.

text: BACKGROUND We previously investigated antibody titers against four kinds of superantigens [streptococcal pyrogenic exotoxin A (SPEA), streptococcal pyrogenic exotoxin C, toxic shock syndrome toxin-1 and staphylococcal enterotoxin B] in patients with Kawasaki syndrome (KS) younger than 6 months of age and reported a relationship between toxic shock syndrome toxin-1 and KS patients. In this study we have investigated antibody titers in KS patients older than 6 months of age. METHODS Serum of 81 patients with KS older than 6 months of age, before intravenous gamma-globulin therapy, and 88 normal age-matched children were used in this study. The IgG antibody titers against four kinds of superantigens were measured with an enzyme-linked immunosorbent assay. RESULTS The KS patients showed significantly elevated mean SPEA titer (P = 0.006) and significantly higher incidence of high SPEA (P = 0.0024) compared with the controls. The SPEA titer in KS patients showed a significant positive correlation with the number of days from onset of illness (P = 0.0002). CONCLUSIONS The elevated antibody titer against superantigens of KS patients older than 6 months of age was different from that of KS patients younger than 6 months of age. Our results suggest that KS patients' exposure to SPEA occurred a few weeks before the onset of KS. SPEA may be one of the possible etiologic agents of KS among patients older than 6 months of age in Kagoshima, Japan.


title: Rabies among African wild dogs (Lycaon pictus) in the Masai Mara, Kenya.

text: A pack of African wild dogs (Lycaon pictus) ranging to the north of the Masai Mara National Reserve in southwestern Kenya was monitored from 1988 to 1989. During a 6-week period (August 1-September 13, 1989), 21 of 23 members of this pack died. Seven carcasses were retrieved, of which 4 were suitable for necropsy and histopathologic examination. Gross findings varied among individuals and included multiple bite wounds, synovitis, lymphadenopathy, submandibular, cervical, and vocal cord edema, blood in bronchi, bronchioles, stomach, and intestine, and interioventral lung lobe consolidation. Histologic examination of 2 available brain samples revealed nonsuppurative encephalitis with eosinophilic intracytoplasmic inclusions (Negri bodies). An additional brain sample tested positive for rabies via a fluorescent antibody test. Other histologic features included severe suppurative bronchopneumonia, myocarditis, and lymphoid depletion of the lymph nodes, tonsils, and spleen. A 304-base pair (bp) nucleotide sequence from the N gene and a 310-bp sequence from the G gene from rabies isolates of 4 wild dogs indicated that infection was with a rabies variant common among domestic dogs in Kenya and Tanzania.


title: Kawasaki disease: diagnosis, management and cardiac sequelae.

text: Kawasaki disease (KD) is an acute self-limiting systemic vasculitis of unknown etiology and the most common cause of acquired coronary disease in children aged 6 months to 5 years. The inflammatory process results in coronary arteritis, aneurysmal lesions, arterial thrombotic occlusion or even sudden death. The diagnostic tests are unknown but treatment with immunoglobulin and aspirin is effective at reducing cardiac complications from 25 to 4.7% in the UK. Myocardial, endocardial or pericardial inflammation may occur acutely or many years later and abnormalities of myocardial blood flow may require ongoing medication, interventional catheterization or even cardiac surgery. There are several new drugs that may have important roles to play in managing KD in children and young adults.


title: Nidovirus sialate-O-acetylesterases: evolution and substrate specificity of coronaviral and toroviral receptor-destroying enzymes.

text: Many viruses achieve reversible attachment to sialic acid (Sia) by encoding envelope glycoproteins with receptor-binding and receptor-destroying activities. Toroviruses and group 2 coronaviruses bind to O-acetylated Sias, presumably via their spike proteins (S), whereas other glycoproteins, the hemagglutinin-esterases (HE), destroy Sia receptors by de-O-acetylation. Here, we present a comprehensive study of these enzymes. Sialate-9-O-acetylesterases specific for 5-N-acetyl-9-O-acetylneuraminic acid, described for bovine and human coronaviruses, also occur in equine coronaviruses and in porcine toroviruses. Bovine toroviruses, however, express novel sialate-9-O-acetylesterases, which prefer the di-O-acetylated substrate 5-N-acetyl-7(8),9-di-O-acetylneuraminic acid. Whereas most rodent coronaviruses express sialate-4-O-acetylesterases, the HE of murine coronavirus DVIM cleaves 9-O-acetylated Sias. Under the premise that HE specificity reflects receptor usage, we propose that two types of Sias serve as initial attachment factors for coronaviruses in mice. There are striking parallels between orthomyxo- and nidovirus biology. Reminiscent of antigenic shifts in orthomyxoviruses, rodent coronaviruses exchanged S and HE sequences through recombination to extents not appreciated before. As for orthomyxovirus reassortants, the fitness of nidovirus recombinant offspring probably depends both on antigenic properties and on compatibility of receptor-binding and receptor-destroying activities.


title: Indirect transmission of bovine viral diarrhoea virus at calving and during the postparturient period.

text: Two trials were designed to investigate whether bovine viral diarrhoea virus (BVDV) could be transmitted after the birth of persistently infected calves, even if they were removed immediately after birth. In trial 1, 11 calves were actively exposed to fetal fluids and uterine lochia collected from cows that had delivered calves persistently infected with type 1 BVDV. One calf that was exposed to a sample taken on the day of calving seroconverted. In trial 2, six calves were housed in stables where persistently infected calves were being born and then removed immediately from their dams and from the stable unit within two to three hours. One of four calves in close contact with the cows after delivery seroconverted and one of two calves housed within the same stable unit became infected.


title: Purification and concentration of viruses associated with transmissible (coronaviral) enteritis of turkeys (bluecomb).

text: Purification and concentration of viruses contained in pathogenic material was accomplished by sucrose density gradient centrifugation. Electron microscopy was also performed. Viruses recovered from intestinal preparations of both affected and unaffected turkey poults had a density of 1.16 to 1.17. The viruses resembled myxoviruses, paramyxoviruses, or oncornaviruses morphologically. In addition, preparations from bluecomb-affected poults were found to contain viruses indistinguishable from known coronaviruses. No such virus particles were seen in the control preparations. Data are presented to indicate an etiologic relationship between the coronavirus and transmissible enteritis of turkeys.


title: Factors influencing readmission after pancreaticoduodenectomy: a multi-institutional study of 1302 patients.

text: OBJECTIVE AND BACKGROUND Morbidity, mortality, and length of hospital stay after pancreaticoduodenectomy (PD) have significantly decreased over recent decades. Despite this progress, early readmission rates after PD have been reported as high as 50%. Few reports have delineated factors associated with readmission after PD. METHODS The medical records of 6 high-volume institutions were reviewed for patients who underwent PD between 2005 and 2010. Data collection included patient characteristics, medical comorbidities, and perioperative factors. Analysis included readmissions up to 90 days after PD. RESULTS A total of 1302 patients underwent PD across all institutions. The 30-day and 90-day readmission rates were 15% and 19%, respectively. The most common reasons for 30-day readmission included infectious complications (n = 65) and delayed gastric emptying (n = 29). The most common reasons for readmission after 90 days included wound infections and intra-abdominal abscess (n = 75) and failure to thrive (n = 38). On multivariate analysis, factors associated with higher readmission rates included a preoperative diagnosis of chronic pancreatitis, higher transfusion requirements, and postoperative complications including intra-abdominal abscess and pancreatic fistula (all P < 0.02). Factors not associated with higher readmission rates included advanced age, body mass index, cardiovascular/pulmonary comorbidities, diabetes, steroid use, Whipple type (standard vs pylorus preserving PD), preoperative endobiliary stenting, and vascular reconstruction. CONCLUSIONS These multi-institutional data represent a large experience of PD without the biases typically of single center studies. Factors related to infection, nutritional status, and delayed gastric emptying were the most common reasons for readmission after PD. Postoperative complications including pancreatic fistula predicted higher rates of readmission.


title: Method for the quantitative measurement of collecting lymphatic vessel contraction in mice.

text: Collecting lymphatic vessels are critical for the transport of lymph and its cellular contents to lymph nodes for both immune surveillance and the maintenance of tissue-fluid balance. Collecting lymphatic vessels drive lymph flow by autonomous contraction of smooth muscle cells that cover these vessels. Here we describe methods using intravital microscopy to image and quantify collecting lymphatic vessel contraction in mice. Our methods allow for the measurement of the strength of lymphatic contraction of an individual lymphangion in a mouse, which has not yet been demonstrated using other published methods. The ability to study murine collecting lymphatic vessel contraction-using the methods described here or other recently published techniques-allows the field to dissect the molecular mechanisms controlling lymphatic pumping under normal and pathological conditions using the wide variety of molecular tools and genetic models available in the mouse. We have used our methods to study lymphatic contraction in physiological and inflammatory conditions. The methods described here will facilitate the further study of lymphatic function in other pathological conditions that feature lymphatic complications.


title: Development of vaccines for Marburg hemorrhagic fever.

text: Marburg (MARV) and Ebola viruses (EBOV) emerged from the rainforests of Central Africa more than 30 years ago causing outbreaks of severe and, usually, fatal hemorrhagic fever. EBOV has garnered the lion's share of the attention, fueled by the higher frequency of EBOV outbreaks, high mortality rates and importation into the USA, documented in such popular works as the best-selling novel 'The Hot Zone'. However, recent large outbreaks of hundreds of cases of MARV infection in the Democratic Republic of the Congo and Angola with case fatalities approaching 90% dramatically highlight its lethal potential. Although no vaccines or antiviral drugs for MARV are currently available, remarkable progress has been made over the last few years in developing potential countermeasures against MARV in nonhuman primate models. In particular, a vaccine based on attenuated recombinant vesicular stomatitis virus was recently shown to have both preventive and postexposure efficacy.


title: Early Dengue infection and outcome study (EDEN) - study design and preliminary findings.

text: INTRODUCTION Dengue is a major public health problem in Singapore. Age-specific dengue morbidity rates are highest in the young adult population, unlike in many other Southeast Asian countries where dengue is mainly a paediatric disease. Hence, the World Health Organization (WHO) guidelines on dengue diagnosis and management which were developed using the paediatric experiences, may not be suitable for the management of adult dengue infections. MATERIALS AND METHODS The Early DENgue (EDEN) infection and outcome study is a collaborative longitudinal study to investigate epidemiological, clinical, viral and host-specific features of early dengue-infected adults, in an effort to identify new early markers for prognostication. Patients presenting with early undifferentiated fever were included in the study. We carried out an interim analysis to look for early indicators of severe disease. RESULTS During the period of this interim study analysis, 455 febrile patients were recruited. Of these, 133 were confirmed as acute dengue cases based on dengue-specific polymerase chain reaction (PCR) results. There were significant clinical and epidemiological differences between dengue and febrile non-dengue cases. Nine per cent of the dengue cases experienced persistent tiredness, drowsiness and loss of appetite beyond 3 weeks of illness. Quantitation of viral loads using the crossover (Ct) value of real-time RT-PCR correlated with the duration of symptoms. More than half of both primary and secondary dengue cases were hospitalised. There was no dengue-related mortality in this study. CONCLUSION The duration of illness and prolonged symptom duration in 9% of the subjects indicate that the burden of dengue illness is substantially different from other non-dengue febrile illness in our study cohort. Our study also highlights the paucity of early prognostic markers for dengue fever in adults.


title: Isolation and identification of autoagglutinating serogroup O:11 Aeromonas strains in the clinical laboratory.

text: We evaluated the extent to which serogroup O:11 Aeromonas strains could be recovered from both clinical and environmental specimens and the cultural parameters that affected the phenotypic marker (autoagglutination) associated with this group. Of over 200 Aeromonas strains screened, serogroup O:11 was identified only among the phenospecies A. hydrophila and A. sobria and was associated with clinical isolates more frequently than with environmental strains. Blood and wound isolates accounted for almost 50% of all O:11 strains identified. The autoagglutination phenotype associated with O:11 strains could be detected in most commercial liquid media, under a wide range of growth temperatures, and within 15 min of incubation at 100 degrees C. The results suggest that clinical laboratories can recognize this important group of Aeromonas strains by two simple tests.


title: Diagnosis of enzootic pneumonia in Danish cattle: reverse transcription-polymerase chain reaction assay for detection of bovine respiratory syncytial virus in naturally and experimentally infected cattle.

text: A reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for detection of bovine respiratory syncytial virus (BRSV) in lung tissue of naturally and experimentally infected cattle. Primers were selected from the gene coding the F fusion protein, which is relatively conserved among BRSV isolates. The RT-PCR assay was highly specific, it yielded positive reactions only when performed on BRSV-infected cell cultures or tissues. The detection limit of the RT-PCR assay was assessed as 5 TCID50. BRSV was detected in tissues of the respiratory tract and in the tracheobroncheal lymph node of calves euthanized 2-8 days after experimental infection with BRSV, whereas samples of other tissues and samples from mock-infected animals were negative at all time points. Examination of lung samples from 8 different regions of the lungs revealed that although the virus was most often found in the cranioventral lobules, it was frequently present in all lung lobules. Microbiologic examinations of all acute fatal cases of pneumonia (135 animals) in cattle submitted for diagnostic purposes during 1 year revealed that Actinomyces pyogenes (11%), Haemophilus somnus (10%), Pasteurella sp. (7%), and Pasteurella haemolytica (7%) were the most common bacterial agents found in the lungs. BRSV was identified using a conventional antigen enzyme-linked immunosorbent assay (ELISA) in 23 (17%) animals. The established BRSV-specific RT-PCR assay yielded positive results for the same 23 animals. In addition, 10 animals that were negative with the ELISA were positive with the RT-PCR assay. These results indicates that the RT-PCR assay can be a sensitive, reliable alternative to conventional diagnostic procedures.


title: RNA recombination of murine coronaviruses: recombination between fusion-positive mouse hepatitis virus A59 and fusion-negative mouse hepatitis virus 2.

text: It has previously been shown that the murine coronavirus mouse hepatitis virus (MHV) undergoes RNA recombination at a relatively high frequency in both tissue culture and infected animals. Thus far, all of the recombination sites had been localized at the 5' half of the RNA genome. We have now performed a cross between MHV-2, a fusion-negative murine coronavirus, and a temperature-sensitive mutant of the A59 strain of MHV, which is fusion positive at the permissive temperature. By selecting fusion-positive viruses at the nonpermissive temperature, we isolated several recombinants containing multiple crossovers in a single genome. Some of the recombinants became fusion negative during the plaque purification. The fusion ability of the recombinants parallels the presence or absence of the A59 genomic sequences encoding peplomers. Several of the recombinants have crossovers within 3' end genes which encode viral structural proteins, N and E1. These recombination sites were not specifically selected with the selection markers used. This finding, together with results of previous recombination studies, indicates that RNA recombination can occur almost anywhere from the 5' end to the 3' end along the entire genome. The data also show that the replacement of A59 genetic sequences at the 5' end of gene C, which encodes the peplomer protein, with the fusion-negative MHV-2 sequences do not affect the fusion ability of the recombinant viruses. Thus, the crucial determinant for the fusion-inducing capability appears to reside in the more carboxyl portion of the peplomer protein.


The returned traveller.


Open tracheostomy in a suspect severe acute respiratory syndrome (SARS) patient: brief technical communication.


title: Preoperative embolization of hypervascular thoracic, lumbar, and sacral spinal column tumors: technique and outcomes from a single center.

text: The existing literature on preoperative spine tumor embolization is limited in size of patient cohorts and diversity of tumor histologies. This report presents our experience with preoperative embolization of hypervascular thoracic, lumbar, and sacral spinal column tumors in the largest series to date. We conducted a retrospective review of 228 angiograms and 188 pre-operative embolizations for tumors involving thoracic, lumbar and sacral spinal column. Tumor vascularity was evaluated with conventional spinal angiography and was graded from 0 (same as normal adjacent vertebral body) to 3 (severe tumor blush with arteriovenous shunting). Embolic materials included poly vinyl alcohol (PVA) particles and detachable platinum coils and rarely, liquid embolics. The degree of embolization was graded as complete, near-complete, or partial. Anesthesia records were reviewed to document blood loss during surgery. Renal cell carcinoma (44.2%), thyroid carcinoma (9.2%), and leiomyosarcoma (6.6%) were the most common tumors out of a total of 40 tumor histologies. Hemangiopericytoma had the highest mean vascularity (2.6) of all tumor types with at least five representative cases followed by renal cell carcinoma (2.0) and thyroid carcinoma (2.0). PVA particles were used in 100% of cases. Detachable platinum coils were used in 51.6% of cases. Complete, near-complete, and partial embolizations were achieved in 86.1%, 12.7%, and 1.2% of all cases, respectively. There were no new post-procedure neurologic deficits or other complications with long-term morbidity. The mean intra-operative blood loss for the hypervascular tumors treated with pre-operative embolization was 1745 cc. Preoperative embolization of hypervascular thoracic, lumbar, and sacral spine tumors can be performed with high success rates and a high degree of safety at high volume centers.


title: Plasma glucose levels and diabetes are independent predictors for mortality and morbidity in patients with SARS.

text: AIMS To investigate the relationships between a known history of diabetes and ambient fasting plasma glucose (FPG) levels with death and morbidity rates in patients with severe acute respiratory syndrome (SARS). METHODS In this retrospective analysis, the clinical and biochemical characteristics of 135 patients who had died from SARS, 385 survivors of SARS and 19 patients with non-SARS pneumonia were compared. RESULTS All patients were treated according to a predefined protocol. Before steroid treatment, the mean FPG level was significantly higher in the SARS group (deceased vs. survivors vs. non-SARS pneumonia group: 9.7 +/- 5.2 vs. 6.5 +/- 3.0 vs. 5.1 +/- 1.0 mmol/l, P < 0.01). In the SARS group, the percentage of patients with a known history of diabetes was significantly higher in the deceased patients than in the survivors (21.5% vs. 3.9%, P < 0.01). Among patients with no known history of diabetes and before commencement of steroid therapy, those who had hypoxaemia (SaO(2) < 93%) had higher FPG levels than those who did not have hypoxia in both the survivor (8.7 +/- 4.9 vs. 6.3 +/- 2.1 mmol/l, P < 0.001) and deceased (9.8 +/- 4.8 vs. 7.2 +/- 1.5 mmol/l, P < 0.001) groups. A known history of diabetes [odds ratio (OR) 3.0, 95% confidence interval (CI) 1.4, 6.3; P = 0.005] and FPG > or = 7.0 mmol/l before steroid treatment (OR 3.3, 95% CI 1.4, 7.7, P = 0.006) were independent predictors of death. During the course of the illness, FPG levels were negatively associated with SaO(2) (beta =-0.682 +/- 0.305, P = 0.025, general estimation equation model) in SARS patients. Survival analysis showed that FPG was independently associated with an increased hazard ratio (HR) of mortality (HR = 1.1, 95% CI 1.0, 1.1, P = 0.001) and hypoxia (HR = 1.1, 95% CI 1.0, 1.1, P = 0.002) after controlling for age and gender. CONCLUSIONS A known history of diabetes and ambient hyperglycaemia were independent predictors for death and morbidity in SARS patients. Metabolic control may improve the prognosis of SARS patients.


Form and representation in clinical case reports.


title: Positive selection of the TRIM family regulatory region in primate genomes.

text: Viral selection pressure has acted on restriction factors that play an important role in the innate immune system by inhibiting the replication of viruses during primate evolution. Tripartite motif-containing (TRIM) family members are some of these restriction factors. It is becoming increasingly clear that gene expression differences, rather than protein-coding regions changes, could play a vital role in the anti-retroviral immune mechanism. Increasingly, recent studies have created genome-scale catalogues of DNase I hypersensitive sites (DHSs), which demark potentially functional regulatory DNA. To improve our understanding of the molecular evolution mechanism of antiviral differences between species, we leveraged 14 130 DHSs derived from 145 cell types to characterize the regulatory landscape of the TRIM region. Subsequently, we compared the alignments of the DHSs across six primates and found 375 DHSs that are conserved in non-human primates but exhibit significantly accelerated rates of evolution in the human lineage (haDHSs). Furthermore, we discovered 31 human-specific potential transcription factor motifs within haDHSs, including the KROX and SP1, that both interact with HIV-1 Importantly, the corresponding haDHS was correlated with antiviral factor TRIM23 Thus, our results suggested that some viruses may contribute, through regulatory DNA differences, to organismal evolution by mediating TRIM gene expression to escape immune surveillance.


title: RNA isolation and quantitative PCR from HOPE- and formalin-fixed bovine lymph node tissues.

text: The use of ribonucleic acid (RNA) extracted from Hepes glutamic acid buffer-mediated organic solvent protection effect (HOPE)-fixed tissues in quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is fairly novel. We compared qRT-PCR analysis of formalin- and HOPE-fixed, paraffin-embedded lymph node tissues from Mycobacterium bovis-infected cattle by extracting total RNA using a commercial kit (Ambion) and a Trizol method. RNA extracted from HOPE-fixed tissues showed comparable quantities between the commercial kit (82.7-107.9 microg/ml total RNA) and the Trizol method (87-161.1 microg/ml total RNA), displaying a high degree of integrity when analyzed by electrophoresis. RNA extracted from formalin-fixed tissues using the commercial kit produced similar concentrations (80.6-145.7 microg/ml total RNA) in comparison to the HOPE tissue; however, the integrity was compromised. Extraction of RNA from the formalin-fixed tissues using Trizol was unsuccessful. Following qRT-PCR for glyceraldehyde-3-phosphate dehydrogenase (GAPDH), total RNA from HOPE-fixed tissues showed higher levels of target messenger ribonucleic acid (mRNA) (4.05 x 10(-2)pg/100 ng total RNA using the commercial kit and 6.45 x 10(-2)pg/100 ng total RNA using Trizol) in comparison to formalin-fixed tissues (5.69 x 10(-4)pg/100 ng total RNA). This could be attributed to RNA degradation by exposure to formalin fixation. In conclusion, the HOPE fixative proved to be a better source for RNA extraction from cattle lymph nodes and subsequent qRT-PCR.


Do no harm: do thyself no harm.


title: Hospital stays, hospital charges, and in-hospital deaths among infants with selected birth defects--United States, 2003.

text: Birth defects (BDs) are conditions that 1) result from a malformation, deformation, or disruption in one or more parts of the body; 2) are present at birth; and 3) have a serious, adverse effect on health, development, or functional ability. BDs are leading causes of pediatric hospitalizations, medical expenditures, and infant mortality. To estimate national hospital charges and rates of in-hospital deaths for a greater number of specific BDs than estimated in previous reports, investigators at the University of Arkansas for Medical Sciences and CDC used the Healthcare Cost and Utilization Project 2003 Kids' Inpatient Database (KID), developed and distributed by the Agency for Healthcare Research and Quality. KID is a 10% sample of hospital discharges after uncomplicated births and an 80% sample of all other pediatric discharges from 36 participating states. Data are weighted to represent all pediatric hospitalizations in the United States. The investigators analyzed hospital stays during 2003 for newborn infants with any of 35 BDs. This report describes the results of that analysis, which indicated substantial variation among BDs regarding average length of stay, average hospital charge, and the incidence of in-hospital deaths. Average length of stay was greatest for infants with surgically repaired gastroschisis or omphalocele. Average hospital charges were highest for infants with hypoplastic left heart syndrome and common truncus arteriosus. Although anencephaly, trisomy 13, and trisomy 18 were associated with the highest rates of in-hospital death, the largest total numbers of deaths associated with neonatal hospitalizations occurred in infants with diaphragmatic hernia and renal agenesis. Further studies are needed to distinguish outcomes for infants with isolated and multiple defects and to assess longer-term outcomes.


title: Detection of Reovirus type 3 by use of fluorogenic nuclease reverse transcriptase polymerase chain reaction.

text: Reovirus type 3 (Reo-3) can infect numerous rodent species and induces the clinical syndrome 'oily skin disease' in neonatal mice, and is a common contaminant of biological materials. The reverse transcriptase polymerase chain reaction (RT-PCR) assay has proven useful for the detection of Reo-3 in rodents and contaminated biological materials. Fluorogenic nuclease reverse transcriptase polymerase chain reaction assays (fnRT-PCR) combine RT-PCR with an internal fluorogenic hybridization probe, thereby potentially enhancing specificity and eliminating post-PCR processing. Therefore, an fnRT-PCR assay specific for Reo-3 was developed by targeting primer and probe sequences to a unique region of the Reo-3 M3 gene. The fnRT-PCR detected both strains of Reo-3 (Dearing and Abney), but did not detect Reovirus types 1 or 2, other viruses in the family Reoviridae, or other RNA viruses that naturally infect rodents. The fnRT-PCR detected less than 1 fg of target template and detected viral RNA in tissues obtained from mice experimentally infected with Reo-3. The assay also displayed comparable sensitivity when compared to the mouse antibody production test commonly used to detect viral contamination of biological materials. In conclusion, this fnRT-PCR assay offers a potentially high-throughput diagnostic assay for detecting Reo-3 RNA in infected mice and contaminated biological materials.


title: Change in infection control practices and awareness of hospital medical staff in the aftermath of SARS.

text: Severe Acute Respiratory Syndrome (SARS) epidemic illustrated the crucial role of infection surveillance and control measures in the combat of any highly transmissible disease. We conducted an interview survey of 121 medical staff 145 doctors, 46 staff nurses and 30 medical assistants) in a state hospital in Malaysia three months after the end of SARS epidemic (from October to December 2003). Staff was grouped according to those directly involved in the care of suspected SARS patients [S+ group n=41] and those who were not [S- group; n=80]. On hand washing following sneezing, coughing and touching patients, the proportions of medical staff that reported an increase after the SARS crisis were 22.3%, 16.5% and 45.5% respectively. On wearing masks, gloves, and aprons when meeting potentially infectious patients, the proportions that reported an increase were 39.7%, 47.1% and 32.2% respectively. Significantly more staff in S+ than S- group reported these increases. Sixty percent of staff was aware of changes in hospital infection control policies after SARS; 93.4% was aware of notifying procedures, and 81.8% was aware of whom to notify in the hospital. Regarding infection isolation ward, Infectious Control Nurse and Infection Control Committee Chairman in the hospital, the proportions of staff that could correctly name them were 39.7%, 38.3% and 15.7% respectively. Significantly more in S+ than S- group could do so. However, more than half the staff claimed ignorance on the knowledge of infection isolation ward (56.2%), Infection Control Nurse (57.9%) and Chairman (65.3%). Our findings demonstrated that SARS crisis had some positive impact on the infection control practices and awareness of medical staff especially on those with direct SARS involvement. Implications for future control of infectious diseases are obvious.


title: Coding sequence of coronavirus MHV-JHM mRNA 4.

text: A coding sequence at the 5' end of mRNA 4 of the coronavirus MHV-JHM was determined by M13/chain-terminator sequencing of cloned cDNA. An open reading frame of 417 bases with the potential to encode a polypeptide of mol. wt. 15 200 (139 residues) was identified. The 3' end of the open reading frame overlapped by 16 bases the start of an open reading frame found in mRNA 5. The translation product of mRNA 4 was predicted to be a basic polypeptide rich in threonine. It had a large hydrophobic region near the amino terminus and a basic carboxy terminus. An intracellular, virus-specific polypeptide, which has been previously described having a mol. wt. of 14 000 to 14 500 has the size and charge characteristics of such a translation product.


title: Massive Pulmonary Thromboembolism in Patients with COVID-19; Report of Three Cases.

text: COVID-19 is a novel infectious disease, which has challenged people all around the world. As of today, healthcare practitioners and researchers have made great effort to understand the characteristics and clinical presentations of the disease; however, the existing literature is still incomplete in this regard. A growing body of evidence indicates that coagulopathies and thromboembolic events are of utmost importance in COVID-19 patients and are related to poor prognosis. Here, we report three ICU admitted cases of COVID-19, in which massive pulmonary thromboembolism (PTE) occurred a few days after disease onset. Unfortunately, one of the patients did not survive and two were treated; one with thrombectomy and other with antithrombotic agents. It seems that severe cases of COVID-19 are at risk for developing PTE and in-charge physicians should be prepared and plan for anticoagulant prophylaxis using low-molecular-weight heparin (LMWH).


title: Seroepidemiologic study of human coronavirus OC43 infections in Italy.

text: Human Coronavirus OC43 from suckling mouse brain (SMB) suspension has been adapted to grow in human embryonic lung fibroblast cell cultures. Complement-fixing and hemagglutinating antigens were prepared from infected cell cultures and used in parallel with the same antigens from SMB for a study of the prevalence of OC43 antibody in Northern Italy as well as for the serodiagnosis of recent OC43 infections and reinfections. Results show (i) a higher sensitivity of the hemagglutination-inhibition as compared to the complement fixation test both for antibody detection in seroepidemiological studies and serodiagnosis of acute infections, (ii) a high prevalence of OC43 antibody in Northern Italy, so that only exceptionally are people older than 12 years seronegative, (iii) a high rate of OC43 reinfections during the winter-spring season 1976/77 in a population sample with preexisting antibody (iv) a good quality of reagents prepared from cell cultures, and hence free of contaminating mouse viruses, such as mouse hepatitis virus.


title: Treatment of de novo femoro-popliteal lesions with a new Drug Coated Balloon: early experience of a single Center in the first 50 patients.

text: Angioplasty with drug-coated balloon (DCB) is an emerging and reliable method for the treatment of femoro-popliteal lesions. We report our experience with the Stellarex™ DCB in the first 50 patients. Methods From July 2015 to November 2017, 50 patients (41 M, 9F), medium age (64 ± 7.4 year) were subject to 33 angioplasties (PTAs) for femoro-popliteal lesions with a paclitaxel-coated balloon (Stellarex™). Based upon clinical data sixteen patients had severe claudication (56% - Rutherford class 3); ten patients suffered from ischemic rest pain (34% - Rutherford class 4); and five presented minor tissue loss (10% - Rutherford class 5). 42% of patients showed femoro-popliteal lesion TASC-II B, and 58% presented lesions pertaining to TASC-II C. Results Immediate technical success was 100% without perioperative complications. Primary patency rate was 94% at twelve months. In three cases restenosis (6%) was detected within a year from procedure, and a further PTA DCB was performed with primary assisted patency rates of 100% at twelve months. Two patients underwent major lower limb amputation. Three patients died during follow-up and one patient was lost at follow-up. Conclusion DCB angioplasty with Stellarex™ is a viable alternative to traditional endovascular procedures proving satisfactory primary patency rates at twelve months. Based on our experience, treatment with DCB is a first choice technique for non-complex de novo lesions of the femoro - popliteal tract.


title: What are the chances? Evaluating risk and benefit information in consumer health materials.

text: Much consumer health information addresses issues of disease risk or treatment risks and benefits, addressing questions such as "How effective is this treatment?" or "What is the likelihood that this test will give a false positive result?" Insofar as it addresses outcome likelihood, this information is essentially quantitative in nature, which is of critical importance, because quantitative information tends to be difficult to understand and therefore inaccessible to consumers. Information professionals typically examine reading level to determine the accessibility of consumer health information, but this measure does not adequately reflect the difficulty of quantitative information, including materials addressing issues of risk and benefit. As a result, different methods must be used to evaluate this type of consumer health material. There are no standard guidelines or assessment tools for this task, but research in cognitive psychology provides insight into the best ways to present risk and benefit information to promote understanding and minimize interpretation bias. This paper offers an interdisciplinary bridge that brings these results to the attention of information professionals, who can then use them to evaluate consumer health materials addressing risks and benefits.


title: Lesions in the small intestine of newborn pigs inoculated with porcine, feline, and canine coronaviruses.

text: The infectivity and pathogenicity to newborn pigs of antigenically related coronaviruses from pigs (transmissible gastroenteritis virus; TGEV), cats (feline infectious peritonitis virus; FIPV), and dogs (canine gastroenteritis virus; CGEV) were studied by light, scanning electron, and immunofluorescence microscopy. Hysterectomy-derived, 12-hour-old pigs were orally given tissue culture or frozen preparations of 6 coronavirus strains (3 porcine, 2 feline, and 1 canine). The pigs were killed at regular intervals between 24 and 144 hours after exposure. Virulent TGEV and virulent FIPV produced necrosis of villous epithelium, resulting in villous atrophy in the jejunum and the ileum. Similar, but less extensive and severe lesions, were produced by the 4 other viruses. Coronaviral antigens were identified by immunofluorescence in villous epithelial cells of pigs that had been inoculated with virulent TGEV, attenuated TGEV, virulent FIPV, and tissue culture-adapted FIPV. In contrast, coronaviral antigens were not induced by the small plaque variant TGEV and virulent CGEV in the villous epithelium, but rather in cells of the lamina propria and crypt epithelium.


title: A candidate modified-live bovine coronavirus vaccine: safety and immunogenicity evaluation.

text: A modified-live vaccine against the respiratory form of bovine coronavirus (BCoV) infection was developed by progressive attenuation of a respiratory strain (438/06-TN). The vaccine was found to be safe as four colostrum-deprived newborn calves remained healthy after oronasal administration of ten doses of the vaccine. The immunogenicity of the vaccine was assessed by intramuscular injection of one vaccine dose to 30 BCoV-antibody negative 2-3-month-old calves. At 30 days post-vaccination, all vaccinated calves displayed high antibody titres against BCoV. Sequence analysis of the S gene of wild-type and cell-adapted 438/06-TN strain detected 10 nucleotide changes, 9 of which were non-synonymous.


title: Disifin (sodium tosylchloramide) and Toll-like receptors (TLRs): evolving importance in health and diseases.

text: Disifin has emerged as a unique and very effective agent used in disinfection of wounds, disinfection of surfaces, materials and water, and other substances contaminated with almost every type of pathogenic microorganism ranging from viruses, bacteria, fungi and yeast, and, very possibly, protozoan parasites, as well. The major active component of Disifin is tosylchloramide sodium (chloramine T). However, the mechanism by which Disifin suppresses the activities of pathogenic microbial agents remains enigmatic. The molecular mechanisms, and the receptors and the signal transducing pathways responsible for the biological effects of Disifin are largely unknown. Despite considerable advances, enormous investigative efforts and large resources invested in the research on infectious diseases, microbial infection still remains a public health problem in many parts of the world. The exact nature of the pathogenic agents responsible for many infectious diseases, and the nature of the receptors mediating the associated inflammatory events are incompletely understood. Recent advances in understanding the molecular basis for mammalian host immune responses to microbial invasion suggest that the first line of defense against microbes is the recognition of pathogen-associated molecular patterns (PAMPs) by a family of transmembrane pattern-recognizing and signal transducing receptor proteins called Toll-like receptors (TLRs). The TLR family plays an instructive role in innate immune responses against microbial pathogens, as well as the subsequent induction of adaptive immune responses. TLRs mediate recognition and inflammatory responses to a wide range of microbial products and are crucial for effective host defense by eradication of the invading pathogens. Now, recent updates demonstrated the ability of Disifin-derived products, Disifin-Animal and Disifin-Pressant to effectively suppress the progression and activities of Chikungunya fever and that of avian influenza A virus [A/cardialis/Germany/72, H7N1: the agent of a highly pathogenic avian influenza (HPAI)] infection, respectively. Overall, the above findings led me to suggest that Disifin and TLRs may mechanistically overlap in the processes of executing their functions against pathogenic microbial organisms. Thus, elucidating and better understanding of the molecular underpinnings responsible for the biochemical effects of Disifin-products, and the nature and mode of the interaction(s) of Disifin with TLRs in the process of exerting their biological effects may open a novel dimension in the research of infectious diseases, which may provide novel therapeutic targets for the prevention and treatment of a wide range of infectious diseases.


title: COVID-19 Case Detection: Cuba's Active Screening Approach.

text: Meningitis, neuropathy, HIV, dengue-since the 1960s, Cuba has faced its share of epidemics. More recently, Cuban health pro-fessionals tackled domestic outbreaks of H1N1 (2009) and Zika (2016), and worked alongside colleagues from around the world to stem Ebola in West Africa; all three were categorized by WHO as public health emergencies of international concern. In December 2019, China reported its fi rst cluster of pneumo-nia cases, later identifi ed as the novel coronavirus disease COVID-19. In January 2020, Cuban authorities convened a multi-sector working group coordinated by the Ministry of Pub-lic Health (MINSAP) and Civil Defense to tailor its national epi-demic control plan to confront the rapidly-spreading disease. The plan features a national reporting system and database, with standard protocols including early case detection, contact tracing and regularly-scheduled public health messaging. In late January, no fewer than six ministries, plus the National Sports and Recreation Institute, Customs, Immigration and national media outlets, came together to adapt domestic proto-cols and design multi-phase control and response mechanisms to combat the SARS-CoV-2 virus.


An opportunity for change.


Risk of cancer from mobile phones is unclear.


title: The transmembrane domain of a carboxyl-terminal anchored protein determines localization to the endoplasmic reticulum.

text: UBC6 is a C-terminal membrane-anchored (type IV) protein, native to Saccharomyces cerevisiae, where it is found in the endoplasmic reticulum. When expressed in mammalian cells, this novel ubiquitin-conjugating enzyme also localizes to the endoplasmic reticulum. UBC6 lacks a lumenal domain and contains no known endoplasmic reticulum retention signals. Analysis of chimeric proteins in which the cytosolic domain of UBC is linked to a heterologous transmembrane domain, or in which the UBC6 transmembrane domain is appended to an unrelated soluble protein, led to the determination that the transmembrane domain of UBC6 plays a dominant role in its compartmental localization. The basis for the transmembrane domain-mediated subcellular targeting of UBC6 was evaluated by lengthening the wild type UBC6 hydrophobic segment from 17 to 21 amino acids, which resulted in re-targeting to the Golgi complex. A further increase in length to 26 amino acids allowed this modified protein to traverse the secretory pathway and gain expression at the plasma membrane. These findings are consistent with models in which, in the absence of dominant cytosolic or lumenal targeting determinants, proteins may be sorted within the secretory pathway based on interactions between their transmembrane domains and the surrounding lipid bilayer.


title: JHM infections in rats as a model for acute and subacute demyelinating disease.

text: An animal model with different central nervous system (CNS) disease processes associated with demyelination is described which provides a basis to analyse the pathogenetic mechanisms leading to these disorders. Intracerebral infection of rats with the murine coronavirus strain JHM can result in an acute encephalomyelitis with a short incubation period or in subacute to chronic encephalomyelitis occurring after prolonged incubation. The most prominent finding of the latter two diseases consists of typical demyelinated lesions distributed in selected areas of the CNS. The induction of high rates of animals with demyelination depends both on properties of the virus used for infection and host factors such as age and immune status. A high number of rats with demyelination was obtained by intracerebral inoculation of temperature sensitive mutants into suckling rats with maternal JHM antibodies.


title: Deduced amino acid sequence and potential O-glycosylation sites for the bovine coronavirus matrix protein.

text: The nucleotide sequence of the matrix (M) protein gene of the bovine coronavirus (BCV) was determined by sequencing cDNA clones derived from genomic RNA. The gene was found to map at the 5' side of the nucleocapsid protein gene and its sequence predicts a protein of 230 amino acids having a molecular weight of 26,376. The BCV M protein shares extensive sequence homology with the matrix protein of the mouse hepatitis coronavirus (MHV) but differs notably in the amino terminal region external to the virion envelope where BCV apparently uses at least two of its six potential O-glycosylation sites.


title: Right-siting of medical care: role of the internist.

text: General internal medicine is now an essential service, and may in time be the main vehicle of delivery of healthcare to an ageing population, since resources are finite. One model for an equitable system of healthcare delivery may be the integration of General Internal Medicine as the core matrix, around which the various subspecialties deliver quality care. This is now a reality in many hospitals, where all subspecialists serve for varying periods in general medical wards, some even achieving dual accreditation. This promotes integration rather than fragmentation of services. Subspecialties will thrive, for the general workload will also be shared by internists in an equitable fashion. The obvious beneficiaries are the patients, and the health economics will also benefit the funding bodies. The services provided by internists must also be expanded into new fields, e.g. medicine for disasters, so as to promote cost-effective medical care, research and teaching, and also to achieve right-siting of patient care. It must also be emphasised that the specialties remain integral parts of the matrix, so that all departments complement one another, rather than compete with each other. The collegiality engendered is essential for a more congenial workplace, so as to promote staff retention.


Financing and organization of China's health care.


New mental health legislation.


title: Recent progress in the characterization of avian enteric viruses.

text: Despite the importance of the poultry gut, remarkably little is known about the complex gut microbial community. Enteric disease syndromes such as runting-stunting syndrome in broiler chickens and poult enteritis complex in young turkeys are difficult to characterize and reproduce in the laboratory. A great deal of work has been done to characterize the bacterial population in the poultry gut, leading to useful performance-based interventions such as direct-fed microbial preparations. Advances in the application of rapid molecular diagnostics and the advent of the next generation of nucleic acid sequencing have allowed researchers to begin to decipher the microbial community in complex environmental samples. Researchers have made great strides recently in placing names to some of the unknown and undescribed small viruses in the poultry gut such as parvoviruses, picornaviruses, picobirnavirus, and calicivirus. Investigation into the novel avian astroviruses continues, and recent progress has been made in the molecular characterization of the avian rotaviruses. This review will focus on the recent advances that have been made in the discovery, description, and characterization of the multitude of viruses that reside in the poultry gut.


title: TLR2 and MyD88 contribute to Lactobacillus casei extract-induced focal coronary arteritis in a mouse model of Kawasaki disease.

text: BACKGROUND Kawasaki disease is the most common cause of acquired cardiac disease and acute vasculitis in children, targets the coronary arteries, and can occasionally be fatal. The pathogenesis and the molecular mechanisms remain unknown. After injection of Lactobacillus casei cell-wall extract (LCCWE), mice develop a focal coronary arteritis that histopathologically resembles Kawasaki disease, but the mechanism remains unclear. Here, we tested the hypothesis that signaling by Toll-like receptors (TLRs) through their key downstream adaptor molecule myeloid differentiation factor 88 (MyD88) is required for the cellular activation and coronary arteritis produced by LCCWE. METHODS AND RESULTS Bone marrow-derived macrophages from TLR2- or MyD88-deficient mice were unresponsive to LCCWE-induced stimulation. In contrast, macrophages obtained from TLR4-deficient mice produced the same amount of interleukin-6 as macrophages from wild-type mice after stimulation with LCCWE. Intraperitoneal injection of LCCWE produced severe focal coronary arteritis in TLR4(-/-) and C57BL/6 control mice but not in TLR2(-/-) or MyD88(-/-) mice. Collectively, these results indicate that LCCWE is a potent inducer of nuclear factor-kappaB via TLR2 but not TLR4 and that this activation proceeds via the MyD88-dependent signaling pathway. In vivo studies suggest that TLR2(-/-) mice are protected from LCCWE-induced coronary arteritis and that this protection is mediated through the adaptor molecule MyD88. CONCLUSIONS Our results provide important insights into the molecular signaling in this mouse model of coronary arteritis. We show here that LCCWE-induced coronary arteritis is dependent on intact TLR2 and MyD88 signaling.


title: [The prevalence of antibodies to influenza virus and respiratory coronavirus in fattening pigs in Spain].

text: The presence of antibodies to two influenza viruses of the type A (H1N1 and H3N2) and to a porcine respiratory coronavirus was investigated in a study lasting a year. 735 blood serum samples were collected from 79 closed pig fattening farms in the province Segovia (Spain). Hemagglutination inhibition was used with influenza viruses. The percentage of positive results was 78.5% and 62.5% respectively for the serotypes H1N1 and H3N2. A clear reduction in the spread of antibodies was observed in the autumn. The ELISA technique was used with the porcine respiratory coronavirus. As antigen we used the antigenically related transmissible porcine gastroenteritis virus. Using this technique 87% of the sera were positive. Some of these sera with representative ELISA values were confirmed by means of serum neutralisation and radioimmune precipitation of the viral proteins. The incidence of these antibodies remained unchanged over the whole year of the investigation.


title: [Hemagglutination inhibition test for the diagnosis of coronavirus infection in cattle].

text: The main aim of the work was to obtain a viral corpuscle suspension with high hemagglutinating activity to be used as an antigen in HAI tests. The bovine coronavirus SC-1 strain was cultivated on ovine foetal renal cells and gave a suspension with a hemagglutinating titer of 1:64-1:128. A monospecific anti-coronavirus immune serum to be used in the HAI test, too, was prepared by hyperimmunization of a gnotobiotic calf. Its hemagglutinating and neutralizing titers were of 1:2560 and, respectively, 8.5 log. The HAI test was performed using the following reagents: the coronavirus suspension as a hemagglutinating antigen, a 1% suspension of mouse red blood corpuscles, the monospecific anti-coronavirus immune serum as a positive control, and a negative serum. 265 serum samples were investigated, collected from cows and calves in 18 farms where neonatal gastroenteritis was diagnosed. In 72.4% out of these samples there were positive results in the IHA test.


title: Epidemiology of acute lung injury.

text: PURPOSE OF REVIEW Acute lung injury and its extreme manifestation, acute respiratory distress syndrome, complicate a wide variety of serious medical and surgical conditions, only some of which affect the lung directly. Despite recent evidence-based advances in clinical management, acute lung injury and acute respiratory distress syndrome are associated with significant mortality. Detailed epidemiology is essential in guiding the recruitment of patients into trials of new therapeutic interventions, thereby improving outcome and allowing directed allocation of scarce resources. RECENT FINDINGS The incidence of acute lung injury in the United States overall (17-64 per 100,000 person-years) seems to be higher than in Europe, Australia, and other developed countries (17-34 per 100, 000 person-years). The mortality rates for patients with acute respiratory distress syndrome range from 34 to 58%. The hypothesis that pulmonary and extrapulmonary acute respiratory distress syndromes are different disease entities continues to gain momentum. A genetic predisposition to acute respiratory distress syndrome may contribute to its pathogenesis and outcome. SUMMARY Recent epidemiologic studies of the incidence of acute lung injury and acute respiratory distress syndrome have indicated a similar incidence in developed societies, and they confirm that mortality is falling in comparison with a decade ago. The awaited publication of new consensus guidelines for the definition of acute lung injury and acute respiratory distress syndrome may render new studies necessary.


title: Maintenance of pluripotency in mouse embryonic stem cells persistently infected with murine coronavirus.

text: A persistently coronavirus-infected embryonic stem (ES) cell line A3/MHV was established by infecting an ES cell line, A3-1, with mouse hepatitis virus type-2. Although almost all A3/MHV cells were found infected, both A3/MHV and A3-1 cells expressed comparable levels of cell surface differentiation markers. In addition, A3/MHV cells retained the ability to form embryoid bodies. These results suggest that persistent coronavirus infection does not affect the differentiation of ES cells.


title: [Chest pain and dyspnea during the recovery period of COVID-19 pneumonia].

text: A 31-year-old non-smoking male patient admitted with and intubated for COVID-19 pneumonia experienced acute chest pain and dyspnea during his recovery period. He was diagnosed with a pneumothorax based on major bullae formation due to COVID-19. The bullae were not visible after extubation and developed rapidly within a few days.


title: Current status of severe acute respiratory syndrome in China.

text: Severe acute respiratory syndrome (SARS), also called infectious atypical pneumonia, is an emerging infectious disease caused by a novel variant of coronavirus (SARS-associated coronavirus, SARS-CoV). It is mainly characterized by pulmonary infection with a high infectivity and fatality. SARS is swept across almost all the continents of the globe, and has currently involved 33 countries and regions, including the mainland China, Hong Kong, Taiwan, North America and Europe. On June 30, 2003, an accumulative total reached 8450 cases with 810 deaths. SARS epidemic was very rampant in March, April and May 2003 in the mainland of China and Hong Kong. Chinese scientists and healthcare workers cooperated closely with other scientists from all over the world to fight the disease. On April 16, 2003, World Health Organization (WHO) formally declared that SARS-CoV was an etiological agent of SARS. Currently, there is no specific and effective therapy and prevention method for SARS. The main treatments include corticosteroid therapy, anti-viral agents, anti-infection, mechanical ventilation and isolation. This disease can be prevented and controlled, and it is also curable. Under the endeavor of the Chinese Government, medical staffs and other related professionals, SARS has been under control in China, and Chinese scientists have also made a great contribution to SARS research. Other studies in developing new detection assays and therapies, and discovering new drugs and vaccines are in progress. In this paper, we briefly review the current status of SARS in China.


title: Should lCU COVID-19 patients empirically receive therapeutic doses of anticoagulant?

text: not available.


title: From mice to humans - murine intelligence for human CD8+ T cell vaccine design.

text: There has been considerable progress in the design of vaccines capable of safely and effectively inducing CD8(+) T cells for prophylaxis and treatment of chronic infectious diseases and cancer. Much of what is known about CD8(+) T cell-mediated immunity has come from pioneering work in mice; this broad overview discusses recent work in mouse systems where lessons may be drawn for human vaccine development. The areas highlighted include antivector immunity, immunodominance, dendritic cell biology and targeting, the role of Toll-like receptors and their exploitation by novel adjuvants, the role of CD4(+) T cell help, regulatory T cells and, finally, some comments on the different requirements of prophylactic versus therapeutic vaccines.


What you don't know really can hurt you.


title: A review of DNA viral infections in psittacine birds.

text: To date, several DNA viral infections have been reported in psittacine birds. Psittacine beak and feather disease (PBFD) is characterized by symmetric feather dystrophy and loss and development of beak deformities. PBFD is caused by beak and feather virus, which belongs to the Circoviridae, and is the most important infection in psittacine birds worldwide. Avian polyomavirus infection causes acute death, abdominal distention, and feather abnormalities. Pacheco's disease (PD), which is caused by psittacid herpesvirus type 1, is an acute lethal disease without a prodrome. Psittacine adenovirus infections are described as having a clinical progression similar to PD. The clinical changes in psittacine poxvirus-infected birds include serious ocular discharge, rhinitis, and conjunctivitis, followed by the appearance of ulcerations on the medial canthi of the eyes. Internal papillomatosis of parrots (IPP) is a tumor disease characterized by progressive development of papillomas in the oral and cloacal mucosa. IPP has been suggested to caused by papillomavirus or herpesvirus. However, information about these diseases is limited. Here we review the etiology, clinical features, pathology, epidemiology, and diagnosis of these DNA viruses.


Enteric coronavirus in symptomless homosexuals.


title: Identification of two neutralizing regions on the severe acute respiratory syndrome coronavirus spike glycoprotein produced from the mammalian expression system.

text: The Spike (S) protein of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) plays important roles in viral pathogenesis and potentially in the development of an effective vaccine against this virulent infectious disease. In this study, the codon-optimized S gene of SARS-CoV was synthesized to construct DNA vaccine plasmids expressing either the full-length or segments of the S protein. High titer S-specific immunoglobulin G antibody responses were elicited in rabbits immunized with DNA against various segments of the S protein. Two neutralizing domains were identified on the S protein, one at the N terminus (Ser12-Thr535) and the other near the C terminus (Arg797-Ile1192).


International infectious disease surveillance during the London Olympic and Paralympic Games 2012: process and outcomes.


title: Attenuation, safety, and efficacy of an infectious bronchitis virus GA98 serotype vaccine.

text: In 1998, novel strains of infectious bronchitis virus (IBV) were identified in chickens from the southeastern United States and classified as a new serotype designated Georgia 98 (GA98). Because of the widespread nature of the GA98 virus in the southeastern United States and the lack of adequate protection with the DE072 vaccine, we developed a specific vaccine for the GA98 serotype. The GA98/0470/98 isolate of IBV was passaged in embryonating chicken eggs 70 times, and attenuation of the virus was determined in specific-pathogen-free chicks. Pass 70 of the GA98/0470/98 strain of IBV when given at 1 day of age by coarse spray and at 14 days of age in the drinking water at 1 x 10(4.5) 50% embryo infectious dose/bird protected against the homologous GA98 challenge as well as provided good protection against the DE072-type virus. In addition, the vaccine was shown to be adequately attenuated and safe at a 10x dosage.


title: Thrombocytopenia in patients with severe acute respiratory syndrome (review).

text: Severe Acute Respiratory Syndrome (SARS) has been recognized as a new human infectious disease caused by a novel coronavirus (SARS-CoV). Hematological changes in patients with SARS were common, including notably lymphopenia and thrombocytopenia. While the former is the result of decreases in CD4+ or CD8+ T-lymphocytes related to the onset of disease or use of glucocorticoids, the latter may involve a number of potential mechanisms. Although the development of autoimmune antibodies or immune complexes triggered by viral infection may play a significant role in inducing thrombocytopenia, SARS-CoV may also directly infect hematopoietic stem/progenitor cells, megakaryocytes and platelets inducing their growth inhibition and apoptosis. Moreover, the increased consumption of platelets and/or the decreased production of platelets in the damaged lungs are a potential alternative mechanism that can contribute to thrombocytopenia in severe critical pulmonary conditions, which has been rarely revealed and will be discussed.


title: Ligasure hemorrhoidectomy versus Stapled hemorrhoidopexy: a prospective randomized clinical trial.

text: AIM The aim of this study was to compare the outcome of Ligasure hemorrhoidectomy (LH) and Stapled hemorrhoidopexy (SH) for prolapsed hemorrhoids. METHODS Sixty-eight patients with grades III and IV hemorrhoids were randomized into two groups of 34 each, to undergo LH (Group 1) or SH (Group 2). Patient demographics, operative details, postoperative pain score, number of parenteral analgesic injections, hospital stay, and time to return to work were all prospectively collected. Postoperative complications and recurrence of prolapse were also recorded. All patients were regularly followed-up for a total period of 18 months. RESULTS Patient demographics and clinical characteristics were similar between both groups. The mean operating time, postoperative pain score, parenteral analgesics, hospital stay and time off work were not statistically significant between both groups. Likewise, both groups had similar postoperative complications except for a residual prolapse that was observed, at four weeks postoperatively, in eight patients (23.53%) in the SH group as compared to two patients (5.89%) in the LH group (P=0.040). Although hemorrhoid recurrence, at 18 months, was also higher among the SH group as compared to the LH group (11.76% vs. 2.94%, respectively), yet the difference was not statistically significant (P=0.163). CONCLUSION Both LH and SH yield comparable good results and minimal side effects for the treatment of grades III and IV hemorrhoids, with less residual prolapses observed with LH. Further, owing to their low postoperative pain, short hospital stay and rapid return to work, both procedures offer an excellent therapeutic option for prolapsed grades III and IV hemorrhoids.


title: Coronavirus proteins: structure and function of the oligosaccharides of the avian infectious bronchitis virus glycoproteins.

text: The recent finding that the E1 glycoproteins of murine coronaviruses contain only O-linked oligosaccharides suggested that this unusual modification might be a distinguishing feature of coronaviruses and might play an essential role in the life cycle of this family of viruses. To examine these possibilities, we analyzed the oligosaccharide moieties of the membrane proteins of the avian coronavirus infectious bronchitis virus. In addition, we determined the effect of inhibiting the glycosylation of these proteins on viral maturation and infectivity. Infectious bronchitis virus virions contain nine proteins. Four of these proteins, GP36, GP31, GP28, and P23, are closely related structurally and appear to be homologous to the E1 proteins of murine coronaviruses. We found that the oligosaccharides of GP31 and GP28 could be removed with endoglycosidase H and that neither of these glycoproteins was detectable in tunicamycin-treated cells. These two results indicated that GP31 and GP28 contain N-linked oligosaccharides. Therefore, O-linked oligosaccharides are not a universal feature of the small coronavirus membrane glycoproteins. Tunicamycin inhibited glycosylation of all of the viral glycoproteins but did not inhibit production of virions by infectious bronchitis virus-infected cells. The virions released by these cells contained only the three non-glycosylated viral proteins P51, P23, and P14. These particles were not infectious. Therefore, it appears that glycosylated infectious bronchitis virus polypeptides are not required for particle formation. However, the viral glycoproteins are apparently indispensible for viral infectivity.


title: Public health systems research: setting a national agenda.

text: The Institute of Medicine has recommended that policy decisions about improvement of national public health systems be guided by sound scientific evidence. However, to date there is no national research agenda to help guide public health systems. The Centers for Disease Control and Prevention was called upon to lead a collaborative consensus-based process to define key research questions and establish a framework to create opportunities to better coordinate, leverage, and identify public health resources, which are increasingly scarce. The public health systems research agenda that emerged from this process has 14 over-arching priority research themes. This national agenda should stimulate and guide research to meet the urgent need to improve the nation's public health systems.


title: Toward prediction of class II mouse major histocompatibility complex peptide binding affinity: in silico bioinformatic evaluation using partial least squares, a robust multivariate statistical technique.

text: The accurate identification of T-cell epitopes remains a principal goal of bioinformatics within immunology. As the immunogenicity of peptide epitopes is dependent on their binding to major histocompatibility complex (MHC) molecules, the prediction of binding affinity is a prerequisite to the reliable prediction of epitopes. The iterative self-consistent (ISC) partial-least-squares (PLS)-based additive method is a recently developed bioinformatic approach for predicting class II peptide-MHC binding affinity. The ISC-PLS method overcomes many of the conceptual difficulties inherent in the prediction of class II peptide-MHC affinity, such as the binding of a mixed population of peptide lengths due to the open-ended class II binding site. The method has applications in both the accurate prediction of class II epitopes and the manipulation of affinity for heteroclitic and competitor peptides. The method is applied here to six class II mouse alleles (I-Ab, I-Ad, I-Ak, I-As, I-Ed, and I-Ek) and included peptides up to 25 amino acids in length. A series of regression equations highlighting the quantitative contributions of individual amino acids at each peptide position was established. The initial model for each allele exhibited only moderate predictivity. Once the set of selected peptide subsequences had converged, the final models exhibited a satisfactory predictive power. Convergence was reached between the 4th and 17th iterations, and the leave-one-out cross-validation statistical terms--q2, SEP, and NC--ranged between 0.732 and 0.925, 0.418 and 0.816, and 1 and 6, respectively. The non-cross-validated statistical terms r2 and SEE ranged between 0.98 and 0.995 and 0.089 and 0.180, respectively. The peptides used in this study are available from the AntiJen database (http://www.jenner.ac.uk/AntiJen). The PLS method is available commercially in the SYBYL molecular modeling software package. The resulting models, which can be used for accurate T-cell epitope prediction, will be made freely available online (http://www.jenner.ac.uk/MHCPred).


Analysis of the cis-acting elements of coronavirus transcription.


Natural killer (NK) cell activity against enteric murine coronavirus mediated by intestinal leukocytes.


title: Evaluation of ProMED-mail as an electronic early warning system for emerging animal diseases: 1996 to 2004.

text: OBJECTIVE To identify emerging animal and zoonotic diseases and associated geographic distribution, disease agents, animal hosts, and seasonality of reporting in the Program for Monitoring Emerging Diseases (ProMED)-mail electronic early warning system. DESIGN Retrospective study. SAMPLE POPULATION 10,490 disease reports. PROCEDURES Descriptive statistics were collated for all animal disease reports appearing on the ProMED-mail system from January 1, 1996, to December 31, 2004. RESULTS Approximately 30% of reports concerned events in the United States; reports were next most common in the United Kingdom, Canada, Australia, Russia, and China. Rabies, bovine spongiform encephalopathy, and anthrax were reported consistently over the study period, whereas avian influenza, Ebola virus, and Hantavirus infection were reported frequently in approximately half of the study years. Reports concerning viral agents composed more than half of the postings. Humans affected by zoonotic disease accounted for a third of the subjects. Cattle were affected in 1,080 reports, and wildlife species were affected in 825 reports. For the 10,490 postings studied, there was a retraction rate of 0.01 and a correction rate of 0.02. CONCLUSIONS AND CLINICAL RELEVANCE ProMED-mail provided global coverage, but gaps in coverage for individual countries were detected. The value of a global electronic reporting system for monitoring emerging diseases over a 9-year period illustrated how new technologies can augment disease surveillance strategies. The number of animal and zoonotic diseases highlights the importance of animals in the study of emerging diseases.


title: Laparoscopic transperitoneal nephrectomy in non-functioning inflammatory kidneys with or without renal stone.

text: OBJECTIVE To compare our laparoscopic simple nephrectomy results in non-functioning inflammatory kidneys with or without renal stones. PATIENTS AND METHODS Patients, who underwent laparoscopic transperitoneal nephrectomy for non-functioning kidney between June, 2010 and October, 2014 were included to study. Overall, data of 32 patients including 15 patients with renal stone (Group 1) and 17 patients without renal stone (Group 2) were retrospectively reviewed. RESULTS Mean age was 44.4 ± 18.5 years (10-71) in group 1 and 35.2 ± 21 years (9-77) in group 2. Mean operation time was 95.0±25.9 minutes (70-175) in group 1 and 86.7 ± 15.1 minutes (70-125) in group 2. Mean estimated blood loss was found to be 64.13 ± 26.67 ml (30-120) in group 1 and 58.94 ± 24.24 ml (30-100) in group 2. Both groups had inflammatory findings in pathological analysis. There was no significant difference between groups regarding estimated blood loss, operation time, pre-operative and post-operative hemoglobin values, percent hemoglobin decrease, complications and hospitalization times (p > 0.05). CONCLUSIONS Non-functioning kidney with or without renal stone could be operated safely with comparable complication rates and success via laparoscopy in experienced hands.


title: Brucella melitensis and Mycobacterium tuberculosis depict overlapping gene expression patterns induced in infected THP-1 macrophages.

text: Pathogens infecting mammalian cells have developed various strategies to suppress and evade their hosts' defensive mechanisms. In this line, the intracellular bacteria that are able to survive and propagate within their host cells must have developed strategies to avert their host's killing attitude. Studying the interface of host-pathogen confrontation can provide valuable information for defining therapeutic approaches. Brucellosis, caused by the Brucella strains, is a zoonotic bacterial disease that affects thousands of humans and animals around the world inflicting discomfort and huge economic losses. Similar to many other intracellular dwelling bacteria, infections caused by Brucella are difficult to treat, and hence any attempt at identifying new and common therapeutic targets would prove beneficial for the purpose of curing infections caused by the intracellular bacteria. In THP-1 macrophage infected with Brucella melitensis we studied the expression levels of four host's genes, i.e. EMP2, ST8SIA4, HCP5 and FRMD5 known to be involved in pathogenesis of Mycobacterium tuberculosis. Our data showed that at this molecular level, except for FRMD5 that was downregulated, the other three genes were upregulated by B. melitensis. Brucella melitensis and M. tuberculosis go through similar intracellular processes and interestingly two of the investigated genes, i.e. EMP2 and ST4SIA8 were upregulated in THP-1 cell infected with B. melitensis similar to that reported for THP-1 cells infected with M. tuberculosis. At the host-pathogen interaction interface, this study depicts overlapping changes for different bacteria with common survival strategies; a fact that implies designing therapeutic approaches based on common targets may be possible.


title: Characterization of a coronavirus isolated from a diarrheic foal.

text: A coronavirus was isolated from feces of a diarrheic foal and serially propagated in human rectal adenocarcinoma (HRT-18) cells. Antigenic and genomic characterizations of the virus (isolate NC99) were based on serological comparison with other avian and mammalian coronaviruses and sequence analysis of the nucleocapsid (N) protein gene. Indirect fluorescent-antibody assay procedures and virus neutralization assays demonstrated a close antigenic relationship with bovine coronavirus (BCV) and porcine hemagglutinating encephalomyelitis virus (mammalian group 2 coronaviruses). Using previously described BCV primers, the N protein gene of isolate NC99 was amplified by a reverse transcriptase PCR (RT-PCR) procedure. The RT-PCR product was cloned into pUC19 and sequenced; the complete N protein of NC99 (446 amino acids) was then compared with published N protein sequences of other avian and mammalian coronaviruses. A high degree of identity (89.0 to 90.1%) was observed between the N protein sequence of NC99 and published sequences of BCV (Mebus and F15 strains) and human coronavirus (strain OC43); only limited identity (<25%) was observed with group 1 and group 3 coronaviruses. Based on these findings, the virus has been tentatively identified as equine coronavirus (ECV). ECV NC99 was determined to have close antigenic and/or genetic relationships with mammalian group 2 coronaviruses, thus identifying it as a member of this coronavirus antigenic group.


title: [Perinatal and neonatal management plan for prevention and control of 2019 novel coronavirus infection (1st Edition)].

text: Since December 2019, the novel coronavirus (2019-nCoV) infection has been prevalent in China. Due to immaturity of immune function and the possibility of mother-fetal vertical transmission, neonates are particularly susceptible to 2019-nCoV. The perinatal-neonatal departments should cooperate closely and take integrated approaches, and the neonatal intensive care unit should prepare the emergency plan for 2019-nCoV infection as far as possible, so as to ensure the optimal management and treatment of potential victims. According to the latest 2019-nCoV national management plan and the actual situation, the Working Group for the Prevention and Control of Neonatal 2019-nCoV Infection in the Perinatal Period of the Editorial Committee of Chinese Journal of Contemporary Pediatrics puts forward recommendations for the prevention and control of 2019-nCoV infection in neonates.


Infection risk puts the brakes on Canada's biomedical research.


title: Non-structural proteins 2 and 3 interact to modify host cell membranes during the formation of the arterivirus replication complex.

text: The replicase polyproteins of equine arteritis virus (EAV; family Arteriviridae, order Nidovirales) are processed by three viral proteases to yield 12 non-structural proteins (nsps). The nsp2 and nsp3 cleavage products have previously been found to interact, a property that allows nsp2 to act as a co-factor in the processing of the downstream part of the polyprotein by the nsp4 protease. Remarkably, upon infection of Vero cells, but not of BHK-21 or RK-13 cells, EAV nsp2 is now shown to be subject to an additional, internal, cleavage. In Vero cells, approximately 50% of nsp2 (61 kDa) was cleaved into an 18 kDa N-terminal part and a 44 kDa C-terminal part, most likely by a host cell protease that is absent in BHK-21 and RK-13 cells. Although the functional consequences of this additional processing step are unknown, the experiments in Vero cells revealed that the C-terminal part of nsp2 interacts with nsp3. Most EAV nsps localize to virus-induced double-membrane structures in the perinuclear region of the infected cell, where virus RNA synthesis takes place. It is now shown that, in an expression system, the co-expression of nsp2 and nsp3 is both necessary and sufficient to induce the formation of double-membrane structures that strikingly resemble those found in infected cells. Thus, the nsp2 and nsp3 cleavage products play a crucial role in two processes that are common to positive-strand RNA viruses that replicate in mammalian cells: controlled proteolysis of replicase precursors and membrane association of the virus replication complex.


title: Molecular mimicry between Fc receptors and viral antigens.

text: Molecular mimicry has been characterized as the presence of common epitopes, either linear or conformational, shared by host and microbial determinants. Such cross-reactivity may lead to an autoimmune disease. On the other hand molecular mimicry between certain viral proteins and host determinant may protect invading virus to be eliminated by immune system and may promote persistence. In this mini-review I discuss the molecular mimicry of S peplomer protein of mouse hepatitis virus, strain JHM (MHV-JHM) to the host Fc gamma receptor (Fc gamma R). MHV-JHM induces in rodents acute encephalomyelitis and surviving animals develop demyelinating disease with concomitant persistent infection. We have demonstrated that rabbit IgG, but not is F(ab')2 fragments, monoclonal rat and mouse IgG and the rat 2.4G2 anti-Fc gamma R mab immunoprecipitated natural and recombinant S peplomer protein of several strains of MHV. Furthermore, MHV-JHM infected cells formed rosettes with anti-sheep red blood cell (SRBC) - antibody coated SRBC. The 2.4G2 anti-Fc gamma R mab are able to neutralize several strains of MHV, presumably by binding to S peplomer protein. Therefore, the Fc binding site of S is present on the surface of MHV-infected cells. This molecular mimicry between S peplomer protein of MHV-JHM and Fc gamma R has been extended to other members of Coronaviridae, namely bovine coronavirus and transmissible gastroenteritis virus but not to infectious bronchitis virus. The molecular mimicry of viral antigens to Fc receptors has been described also for members of Herpesviridae, namely Herpes simplex, cytomegalovirus and Varicella zoster.(ABSTRACT TRUNCATED AT 250 WORDS)


Laughter is the best medicine.


title: Postoperative non-steroidal anti-inflammatory drugs and colorectal anastomotic leakage. NSAIDs and anastomotic leakage.

text: Anastomotic leakage (AL) is the most important and one of the most serious complications after colorectal resections with primary anastomosis. Any factors that contribute to increase the risk of AL should be identified and--if possible--eliminated. Non-steroidal anti-inflammatory drugs (NSAIDs) are often used for treating pain after surgical procedures, among these also colorectal resections. The objective of this Ph.d. thesis was to investigate whether the use of NSAIDs in the postoperative period increases the risk of AL, and investigate the effect on pathophysiological mechanisms. In order to achieve this, the following studies were performed. Study I was a retrospective, case-control study in 75 patients undergoing laparoscopic colorectal resection for colorectal cancer. 33 of these patients received the NSAID diclofenac in the postoperative period; the remaining 42 did not receive any NSAID. There were significantly more ALs among the patients receiving diclofenac (7/33 vs. 1/42, p=0.018). In uni- and multivariate logistic regression analyses, diclofenac was the only factor associated with increased AL rate. This study functioned as a hypothesis generating study and laid the ground for the subsequent studies. Study II was an experimental, randomized, case-control study in 32 Wistar rats. The rats had a colonic anastomosis performed and were randomized to diclofenac or placebo treatment. After three days, the rats were sacrificed and the anastomoses were harvested. First, the anastomotic strengths were tested by longitudinal; subsequently, the levels of the enzyme cyclooxygenase-2 (COX-2) in the anastomotic tissues were measured. There was no difference among the groups with regard to anastomotic strength, but the animals treated with diclofenac had significantly lower COX-2 levels (median (range) 1.30 (0.42-3.31) ng/mg vs. 2.44 (0.88 - 18.94) ng/mg, p<0.001). This study showed that the used dose of diclofenac was sufficient and relevant, but did not show a direct damaging effect on the anastomoses due to NSAID treatment. Study III was also an experimental, randomized, case-control study. This time round, 60 Wistar rats were included. Again, colonic anastomoses were performed and the rats were randomized to diclofenac or placebo. Also, expanded polytetrafluoruethylene (ePTFE) tubes were placed under the skin of the rats. In this material, substituents of connective tissue accumulate and the amount of accumulation can be measured. After 7 days, the rats were sacrificed and, again, anastomotic strengths were measured along with collagen content in the ePTFE tubes. Anastomotic strength was similar in the two groups while collagen accumulation was significantly decreased among the rats treated with diclofenac (median (i.q.r.) 0.29 (0.13-0.47) vs. 0.47 (0.28-0.62) mcg/mg, p = 0.03). This study for the first time showed that NSAID inhibit subcutaneous collagen formation and that this formation is reversely correlated to anastomotic strength. This information can be used in further studies in this subject. Study IV was the final experimental case-control study in 40 Wistar rats. This time, in order to more easily extrapolate experimental results to daily clinical life, the colonic anastomoses were sutured with the same type of suture material as used in the clinical setting. Thus, half the anastomoses was performed with resorbable suture; the other half with non-resorbable suture. None of the rats received NSAID. The breaking strength was compared and found similar in the two groups. This study showed that experimental studies can be optimized in order to make comparisons and extrapolations to the clinical setting easier. Study V was a database study based on data from the Danish Colorectal Cancer Group's (DCCG) prospective database and electronically registered medical records. From the database information on demographic, surgical and postoperative variables (including AL) were provided. Information on NSAID consumption was retrieved by individual searches in the patients' medical records. Based on these data, uni- and multivariate logistic regression analyses were performed. These analyses identified NSAID treatment in the postoperative period as an individual risk factor for AL. Other risk factors identified were consistent with the available literature. The detrimental effect of the NSAIDs are possibly due to an effect on collagen metabolism leading to weakened tissue around the anastomosis and/or on the risk of thrombosis formation leading to more thromboses in the vessels supplying the anastomosis, thereby limiting anastomotic blood flow. In conclusion, the studies included in this thesis have elucidated some of the physiological and pathophysiological mechanisms involved in anastomotic healing and leakage, and furthermore have shown that the use of NSAIDs in the postoperative period increase the risk of AL in patients undergoing colorectal surgery with primary anastomosis. Based on the findings in these studies, and based on existing knowledge, it is recommended that NSAIDs be abandoned after colorectal resection with primary anastomosis. It should be investigated whether the NSAIDs are also harmful to other types of anastomoses and after other surgical procedures where early tissue healing is crucial.


The impact of the intracerebral antibody response on the clinical course of a virus-induced demyelination in a rat model system.


title: Respiratory consequences of rhinovirus infection.

text: Rhinoviruses, a genus of the family Picornaviridae, are the cause of more than 50% of respiratory tract infections. Complications of rhinovirus infections, which include otitis media, sinusitis, exacerbations of asthma, and other pulmonary diseases, can be significant in certain populations. Reverse transcriptase-polymerase chain reaction has allowed the identification of rhinoviruses and led to a greater appreciation of the role of this pathogen in upper and lower respiratory tract disease. Furthermore, antiviral agents with broad activity against rhinoviruses have recently been developed, have undergone clinical trials, but have not been approved for clinical use. By altering the clinical course of picornavirus infections, it may be possible to minimize their potential adverse consequences.


title: Emerging zoonoses and pathogens of public health significance--an overview.

text: Emerging zoonotic diseases have assumed increasing importance in public and animal health, as the last few years have seen a steady stream of new diseases, each emerging from an unsuspected quarter and causing severe problems for animals and humans. The reasons for disease emergence are multiple, but there are two main factors--expansion of the human population and globalisation of trade. Current issues such as the increasing movement of a variety of animal species, ecological disruption, uncultivatable organisms, and terrorism, all imply that emerging zoonotic diseases will in all probability, not only continue to occur, but will increase in the rate of their emergence. The recurring nature of the crises dictates that closer integration of veterinary and medical communities is warranted, along with improved education of the general public and policy makers.


title: In vitro characterization of TMPRSS2 inhibition in IPEC-J2 cells.

text: The transmembrane serine protease, TMPRSS2 is an important target in the treatment of seasonal influenza infections and contributes to prostate carcinogenesis and metastasis. In this study, the effect of the synthetic TMPRSS2 inhibitor I-432 on jejunal IPEC-J2 cell monolayers cultured on membrane inserts was characterized. Using a fluorogenic substrate, it was found that the apical addition of I-432 could suppress trypsin-like activity in the supernatants of IPEC-J2 cells. The inhibition of TMPRSS2 did not affect physiologically produced hydrogen peroxide levels in the apical and in basolateral compartments. Loss of expression of the TMPRSS2 serine protease domain (28 kDa) was also observed when cells were pre-exposed to I-432. Partial decrease in immunofluorescent signal intensities derived from the altered distribution pattern of TMPRSS2 was detected after a 48 h long incubation of IPEC-J2 cells with the inhibitor indicating the efficacy of TMPRSS2 inhibition via I-432 administration in vitro.