title: Serological and molecular evidence of enterovirus infection in patients with end-stage dilated cardiomyopathy.

text: OBJECTIVE To study the relative diagnostic value of enterovirus-specific molecular biological and serological assays in patients with end-stage dilated cardiomyopathy, and to investigate the possible role of other cardiotropic viruses in dilated cardiomyopathy. DESIGN Analysis of recipient myocardial tissue and serum from patients with dilated cardiomyopathy and controls undergoing cardiac transplantation for end-stage cardiac disease. SETTING University virology department and transplantation unit. METHODS Reverse transcriptase-polymerase chain reaction and nucleotide sequence analysis of myocardial RNA and DNA; enterovirus-specific in situ hybridization; enterovirus-specific immunoglobulin M detection. RESULTS Enterovirus RNA was detected in myocardial tissue from only a small proportion of (five of 75) hearts. However, although enterovirus-specific immunoglobulin M responses were detected in 22 (28%) of 39 controls patients, a significantly higher prevalence was observed among patients with dilated cardiomyopathy (22 (56%) of 39 patients; P < 0.005). All enteroviruses detected in myocardium showed greatest nucleotide sequence homology with coxsackievirus type B3. Detection of enterovirus RNA in myocardium by the polymerase chain reaction and by in situ hybridisation gave comparable results. Other potentially cardiotropic virus genomes, including human cytomegalovirus, influenzaviruses, and coronaviruses were not detected in myocardium. CONCLUSION This study found that enterovirus-specific immunoglobulin M responses provided the strongest evidence of enterovirus involvement in patients with end-stage dilated cardiomyopathy. However, the high background prevalence of these responses limits their diagnostic value. The finding that enteroviruses detected in myocardium were coxsackievirus type B3 accords with recent findings in patients with acute myocarditis, and indicates that this serotype is the major cardiotropic human enterovirus.


title: Use of IgY antibody to recombinant avian reovirus σC protein in the virus diagnostics.

text: Avian reovirus (ARV) is an important agent of several diseases causing considerable losses in poultry farming. An outer capsid protein (σC) of ARV, is known as a virus-cell attachment protein essential for virus infectivity. In this study, the σC gene of ARV was cloned and expressed in Escherichia coli. The expressed recombinant protein was used as immunogen for raising a specific IgY antibody in laying hens. At 14 weeks post immunization, the antibody titers in serum and egg yolk reached 302,000 and 355,000, respectively. The IgY antibody was capable to neutralize ARV in BHK-21 cells and it strongly reacted in ELISA with ARV but not with heterologous viruses. The IgY antibody detected ARV in field samples of infected animal tissues in dot blot assay. These results suggest that an efficient, economic and rapid diagnostics of ARV can be performed routinely using the IgY antibody against a recombinant ARV σC protein.


title: Severe acute respiratory syndrome.

text: The mysterious severe acute respiratory syndrome (SARS) that has originated from the southern Chinese province of Guangdong appears to be a major public health threat and medical challenge. Laboratory studies of SARS patients in a number of countries identified the etiologic agent being a novel member of coronaviridae. High RNA concentrations of this virus in sputum make it as a highly infectious agent. Low concentrations of viral genome are also detectable in feces. Coronaviruses are ubiquitos. They cause disease in many animals including pigs, cattle, dogs, cats, and chickens. These viruses have been associated with upper respiratory infections and sometimes pneumonia in humans. SARS presents with fever, cough, malaise, dyspnea, and hypoxemia. Chest radiographs from affected regions are associated with progressive airway disease. Clinical laboratory features of SARS include lymphopenia, thrombocytopenia, and elevated lactate dehydrogenase levels. Currently, there is no FDA approved pharmacologic treatment for SARS. To date, no convincing clinical data is available for treatment of SARS with ribavirin. While there are some controversies about the use of systemic corticosteroids, Martin et al, in this issue of MSM, present their views on the use of pentoxyfylline (PTX) as a potential agent to be considered for SARS treatment. Finally, our analytical approach to the risks of SARS will certainly enable us to


title: Clinical brief: severe acute respiratory syndrome (SARS).

text: The World Health Organization (http:/www.who.int/en/) and Centers for Disease Control and Prevention (http://www.cdc.gov) websites are updated daily as new information is learned about the Severe Acute Respiratory Syndrome (SARS) phenomena. The transmission of SARS appears to occur predominantly through direct contact with infectious material. This includes dispersed respiratory droplets, making touching contaminated objects a potential concern. Occupational health nurses must educate themselves and their employees about SARS to allow for accurate and timely sharing of information.


Diaphragmatic Crural Eversion: Mid-Term Data of a Novel Technique to Optimize Hiatoplasty during Laparoscopic Repair of Hiatal Hernia.


title: Biological and molecular differentiation between coronaviruses associated with neonatal calf diarrhoea and winter dysentery in adult cattle.

text: Cytopathic coronaviruses were isolated in HRT-18 cells from bloody faecal samples collected from cows in Québec dairy herds with classical winter dysentery (WD). The formation of polykaryons in the infected cell cultures was found to be dependent on the presence of trypsin in the medium. Virus identification was confirmed by indirect immunofluorescence and indirect protein A-gold immunoelectron microscopy using rabbit hyperimmune serum, as well as monoclonal antibodies directed against the spike (S) and hemagglutinin-esterase (HE) glycoproteins of the prototype Mebus strain of bovine coronavirus (BCV-Meb). Four WD isolates differed from BCV-Meb by their ability to agglutinate rat erythrocytes at 4 and 37 degrees C, their higher receptor destroying enzyme activity, but lower acetylesterase activity. The WD isolates were serologically indistinguishable from the reference BCV-Meb strain by virus neutralization and Western immunoblotting, but could be differentiated by hemagglutination-inhibition. Sequence analysis of the PCR-amplified HE gene of a plaque-purified WD isolate (BCQ-2590) revealed sufficient number of nucleotide and amino acid substitutions which may explain this antigenic variability.


title: Genomic organization and expression of the 3' end of the canine and feline enteric coronaviruses.

text: The genomic organization at the 3' end of canine coronavirus (CCV) and feline enteric coronavirus (FECV) was determined by sequence analysis and compared to that of feline infectious peritonitis virus (FIPV) and transmissible gastroenteritis virus (TGEV) of swine. Comparison of the latter two has previously revealed an extra open reading frame (ORF) at the 3' end of the FIPV genome, lacking in TGEV, now designated ORF 6b. Both CCV and FECV possess 6b-related ORFs. The CCV ORF 6b is colinear with that of FIPV, but the predicted amino acid sequences are only 58% identical. The FECV ORF 6b contains a large deletion compared to that of FIPV, reducing the colinear part to 60%. The sequence homologies were highest between CCV and TGEV on the one hand and between FECV and FIPV on the other. The expression product of the CCV and the FECV ORF 6b can be detected in infected cells by immunoprecipitation.


title: Dynamic changes in blood cytokine levels as clinical indicators in severe acute respiratory syndrome.

text: OBJECTIVE To investigate the dynamic changes observed in serum levels of interleukins (ILs), tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) in severe acute respiratory syndrome (SARS) patients. METHODS Sixty-one cases of SARS with positive antibodies to SARS coronavirus (SARS-CoV) were classified into the following categories: initial stage (3 - 7 days), peak stage (8 - 14 days), and remission and recovery stage (15 - 27 days). Forty-four healthy individuals were used as controls. Serum levels of ILs, TNF-alpha and TGF-beta 1 were measured in all subjects. Serum antibodies to SARS-CoV were detected only in SARS cases. RESULTS The mean concentration of serum IL-6 in SARS patients did not differ from that in the control group in initial and peak stages, but became significantly higher in remission and recovery stage compared with the control group, initial and peak stages (P < 0.01). The mean concentration of serum IL-8 in SARS patients did not differ from that of the control group in initial stage, but was significantly higher than control group in peak stage and remission and recovery stage (P < 0.05). And it was more significantly higher in remission and recovery stage than in peak stage (P < 0.01). The mean concentrations of IL-16 and TNF-alpha in SARS patients were higher than those of the control group for every length of the clinical courses investigated, and were especially high in remission and recovery stage (P < 0.01). SARS patients experienced higher concentration of serum IL-13 compared with the controls in initial stage (P < 0.01), but returned to normal levels in peak stage and in remission and recovery stage. The mean concentration of serum IL-18 in SARS patients was significantly lower than that of the control group during all clinical courses (P < 0.05). The mean concentration of serum TGF-beta1 in SARS patients was higher than that of the control group during all clinical courses. Although TGF-beta1 in serum decreased in remission and recovery stage in SARS patients, the average was still higher than that of the control group (P < 0.01). CONCLUSIONS Most proinflammatory cytokines and TGF-beta 1 were elevated during the early phase of SARS, which may be associated with lung infiltration and proliferation. Concurrently, the mean concentration of serum IL-13 decreased gradually, and the mean concentration of serum IL-18 level in SARS patients was lower than that of the control group during all the courses of SARS, suggesting that the immune state of the patients with SARS was obviously abnormal. Observing the dynamic changes in blood cytokine levels can provide a scientific basis to assess pathogenesis and efficacy of clinical treatment of SARS.


title: Membrane assembly of the triple-spanning coronavirus M protein. Individual transmembrane domains show preferred orientation.

text: The M protein of mouse hepatitis virus strain A59 is a triple-spanning membrane protein which assembles with an uncleaved internal signal sequence, adopting an NexoCcyt orientation. To study the insertion mechanism of this protein, domains potentially involved in topogenesis were deleted and the effects analyzed in topogenesis were deleted and the effects analyzed in several ways. Mutant proteins were synthesized in a cell-free translation system in the presence of microsomal membranes, and their integration and topology were determined by alkaline extraction and by protease-protection experiments. By expression in COS-1 and Madin-Darby canine kidney-II cells, the topology of the mutant proteins was also analyzed in vivo. Glycosylation was used as a biochemical marker to assess the disposition of the NH2 terminus. An indirect immunofluorescence assay on semi-intact Madin-Darby canine kidney-II cells using domain-specific antibodies served to identify the cytoplasmically exposed domains. The results show that each membrane-spanning domain acts independently as an insertion and anchor signal and adopts an intrinsic preferred orientation in the lipid bilayer which corresponds to the disposition of the transmembrane domain in the wild-type assembled protein. These observations provide further insight into the mechanism of membrane integration of multispanning proteins. A model for the insertion of the coronavirus M protein is proposed.


Reflections on 45 years as a clinical hematologist.


title: Vaccinia as a vector for gene delivery.

text: Gene therapy is a promising approach, yet so far it has shown limited effectiveness in many clinical trials, mainly due to insufficient gene transduction. Recombinant vaccinia virus (rVV) has been well developed as a gene delivery vector, initially for protein expression in mammalian cells. rVV has been further developed to express antigens in vivo in generating immunity for protection against specific infectious diseases and cancer. rVVs, as non-replicating viral vectors, have been demonstrated for their great potential as vaccines, for their diminished cytopathic effects, high levels of protein expression and strong immunogenicity, and they are relatively safe in animals and in human patients. A number of clinical trials using rVVs as vaccines have shown promising results for treating infectious diseases and cancer. In the last few years, due to its exceptional ability to replicate in tumour cells, the Western Reserve strain vaccinia has been explored as a replicating oncolytic virus for cancer virotherapy. As more is learned about the functions of viral gene products in controlling the mammalian cell cycle and in disabling cellular defence mechanisms, specific viral functions can be augmented or eliminated to enhance antitumour efficacy and improve tumour cell targeting. General mechanisms by which this oncolytic virus achieves the antitumour efficacy and specificity are reviewed. Specifically, the deletion of the viral genes for thymidine kinase and vaccinia growth factor resulted in a vaccinia mutant with enhanced tumour targeting activity and fully retaining its efficiency of replication in cancer cells. Other potential strategies for improving this vector for gene delivery will also be discussed in this review.


title: Quantification and characterization of pleural fluid in healthy dogs with thoracostomy tubes.

text: OBJECTIVE To quantify and characterize pleural fluid collected from healthy dogs after placement of a thoracostomy tube (TT). ANIMALS 8 healthy Coonhound-cross dogs (mean ± SD weight, 27.2 ± 1.6 kg). PROCEDURES Thoracic CT of each dog was performed before placement of a TT and daily thereafter for 7 days. Thoracic fluid volume was calculated from CT images. Effusion was aspirated when detected; volume was recorded, and cytologic analysis and bacterial culture were performed. RESULTS Mean ± SD volume of pleural effusion detected by CT was 1.43 ± 0.59 mL/kg (range, 0.12 to 3.32 mL/kg). Mean volume collected via aspiration was 0.48 ± 0.84 mL/kg (range, 0 to 2.16 mL/kg). Cytologic analysis yielded results consistent with an exudate, characterized by septic suppurative inflammation in 6 dogs and mixed inflammation in 1 dog; there was insufficient volume for analysis in 1 dog. Sufficient volume was obtained for bacterial culture for 6 dogs, which yielded pure growths of Staphylococcus pseudintermedius (n = 3) and Streptococcus equi subspecies zooepidemicus (2) and mixed growth of both of these species (1). The TT was removed before day 7 in 4 dogs because of pyothorax (n = 3) and irreversible damage to the TT (1). CONCLUSIONS AND CLINICAL RELEVANCE Presence of a TT induced a minimal volume of pleural effusion in healthy dogs. Pyothorax developed in most dogs between 4 and 6 days after TT placement. On the basis of these findings, a TT should be removed by the fourth day after placement, unless complications are detected sooner.


title: Fluid administration, vasopressor use and patient outcomes in a group of high-risk cardiac surgical patients receiving postoperative goal-directed haemodynamic therapy: a pilot study.

text: The role of goal-directed therapy in high-risk cardiac surgical patients has not been determined. This study sought to observe the effect of a postoperative standardised haemodynamic protocol (SHP) on the administration of fluid and vasoactive drugs after high-risk cardiac surgery. This was an interventional pilot study. In 2010 to 2011, the SHP was introduced to the ICU at Wellington Regional Hospital, Wellington, New Zealand, for the perioperative management of patients undergoing high-risk cardiac surgery. A pulmonary artery catheter was inserted in the patients in the study group and fluids and supportive medications were provided in the ICU according to a protocol that targeted a cardiac index ≥ 2 l/min/m², mixed venous oxygen saturation ≥ 60% and a mean arterial pressure of 65 to 75 mmHg. Data from 40 consecutive high-risk cardiac surgical patients assigned to this protocol were compared with a matched cohort of 40 consecutive high-risk cardiac surgical patients receiving 'usual care' in 2009. Baseline characteristics were similar in the two groups. There was no significant difference in the duration of noradrenaline infusion in the SHP cohort compared to historical controls (median [IQR] 18.5 hours [31.63] versus 18 hours [18.3]; P=0.35), despite patients receiving more fluid in their first 12 hours in the ICU (mean 4687 ml [SD ± 2284 ml] versus 1889 ml [SD ± 1344 ml]; P <0.001). The SHP cohort had a higher rate of reintubation (4 in 37 [10.8%] versus 0 in 40 [0%]; P=0.049). The SHP delivered significantly more fluid, but did not reduce the duration of noradrenaline infusion, compared to usual care.


title: [Some medical staff positive for serum SARS coronavirus antibody IgG have only mild symptoms].

text: OBJECTIVE To identify patients with SARS coronavirus infection who have only mild symptoms. METHOD Enzyme-linked immunosorbent assay was employed to detect serum antibody against SARS coronavirus in the lysate of whole SARS coronavirus from 19 SARS patients and 200 medical staff members without obvious SARS symptoms after possible exposure to the virus during routine medical practice. RESULTS Serum IgG antibody against SARS coronavirus was detected in all the 19 SARS patients, and among the 200 staff members, 20 (10%) were found positive for the antibody but with no obvious or only mild symptoms. CONCLUSION Serum IgG antibody against SARS coronavirus is positive in a small proportion (around 10%) of the medical staff members exposed to the virus in our hospital, but may not cause obvious symptoms, suggesting SARS coronavirus infection might in some cases have mild or even no clinical manifestations.


title: Hepatobiliary pathology.

text: PURPOSE OF REVIEW Recent papers on disorders of the liver and biliary tract which clarify their pathogenesis and attendant morphologic changes are highlighted. RECENT FINDINGS The concept of 'bystander hepatitis' was cited in studies showing hepatic infiltration of CD8-positive T cells in the setting of extrahepatic infections such as influenza virus and severe acute respiratory syndrome. Diabetic liver lesions include glycogenic hepatopathy (in which poor diabetic control leads to swollen, glycogen-filled hepatocytes without fat, steatohepatitis or fibrosis) and diabetic hepatosclerosis in which there is diffuse perisinusoidal fibrosis (type IV collagen) without zonal predilection. Ground-glass hepatocellular inclusions (positive with periodic acid-Schiff stain for glycogen) were reported in three separate series of patients who were hepatitis B virus-negative, often transplant recipients, immunosuppressed and on multiple medications. A Banff consensus paper expertly compared and contrasted the histologic features which characterize the various causes of late liver allograft dysfunction. SUMMARY Informative papers emerged this past year concerning collateral damage to the liver in extrahepatic infections, diabetic lesions and causes of liver dysfunction after transplantation, among other topics.


title: Anosmia: diagnosis and management.

text: Disorders of the sense of smell can be frustrating for both the patient and physician. Ongoing research in this field has provided insight into the possible mechanisms for smell loss; however, therapy is still limited. Commercially distributed smell testing kits and newer screening tests using material available in all clinical settings have made diagnosis and measurement of the degree of impairment available to all physicians. A detailed history and physical examination are the most powerful tools in the evaluation of smell disorders, whereas imaging studies are reserved for preoperative planning or detailed assessment of positive physical findings.


title: Clinical aspects of immunosuppression in poultry.

text: Chickens, turkeys, and other poultry in a production environment can be exposed to stressors and infectious diseases that impair innate and acquired immunity, erode general health and welfare, and diminish genetic and nutritional potential for efficient production. Innate immunity can be affected by stressful physiologic events related to hatching and to environmental factors during the first week of life. Exposure to environmental ammonia, foodborne mycotoxins, and suboptimal nutrition can diminish innate immunity. Infectious bursal disease (IBD), chicken infectious anemia (CIA), and Marek's disease (MD) are major infectious diseases that increase susceptibility to viral, bacterial, and parasitic diseases and interfere with acquired vaccinal immunity. A shared feature is lymphocytolytic infection capable of suppressing both humoral and cell-mediated immune functions. Enteric viral infections can be accompanied by atrophic and depleted lymphoid organs, but the immunosuppressive features are modestly characterized. Some reoviruses cause atrophy of lymphoid organs and replicate in blood monocytes. Enteric parvoviruses of chickens and turkeys merit further study for immunosuppression. Hemorrhagic enteritis of turkeys has immunosuppressive features similar to IBD. Other virulent fowl adenoviruses have immunosuppressive capabilities. Newcastle disease can damage lymphoid tissues and macrophages. Avian pneumovirus infections impair the mucociliary functions of the upper respiratory tract and augment deeper bacterial infections. Recognition of immunosuppression involves detection of specific diseases using diagnostic tests such as serology, etiologic agent detection, and pathology. Broader measurements of immunosuppression by combined noninfectious and infectious causes have not found general application. Microarray technology to detect genetic expression of immunologic mediators and receptors offers potential advances but is currently at the developmental state. Control methods for immunosuppressive diseases rely largely on minimizing stress, reducing exposure to infectious agents through biosecurity, and increasing host resistance to infectious immunosuppressive diseases by vaccination. A longer-term approach involves genetic selection for resistance to immunosuppressive diseases, which has shown promising results for MD but equivocal results for IBD and CIA.


Role for primary care in epidemic surge capacity.


Application of non-invasive ventilation in China over 20 years.


title: The 3a accessory protein of SARS coronavirus specifically interacts with the 5'UTR of its genomic RNA, Using a unique 75 amino acid interaction domain.

text: More than four years have passed since the outbreak of the severe acute respiratory syndrome (SARS) epidemic, and still very little is known about the molecular biology and pathogenesis of this deadly virus. Among the accessory proteins of the SARS coronavirus (SARS-CoV), the 3a protein has been shown to interact with the spike, envelope, and membrane glycoprotein and has recently been established to be a structural component of capsid. Recent studies suggest that the 3a protein may function as an ion channel and may promote virus release. In order to further characterize the functional properties of this protein, we initiated studies to check its RNA binding activity. Using the yeast three-hybrid system, electrophoretic mobility shift assay (EMSA), and ultraviolet (UV) cross-linking techniques, we have shown that the 3a protein is capable of binding specifically to the 5' untranslated region (5'UTR) of the SARS virus genomic RNA. Further, we have mapped the interaction domain of the 3a protein responsible for this RNA-protein interaction using a series of deletion mutants and defined it to the central 75 amino acid region. This RNA binding motif of 3a does not share homology with any other known RNA binding protein and may have an important role in viral capsid assembly and pathogenesis.


title: Should patients stop their radiotherapy or chemotherapy during the COVID-19 pandemic.

text: In the previous stage, there were too many patients with Corona virus disease 2019 (COVID-19) in Wuhan. Ordinary people, patients, even doctors, had a great sense of desperate. On the one hand, almost all doctors participated in the treatment of patients of COVID-19. On the other hand, the government restricted residents to go out, and the sick people were also afraid of being infected with COVID-19 when seeking medical treatment. Whether cancer patients seek medical treatment or not has become a contradiction for a long time. Our Viewpoint paper is to provide a positive signal to doctors and patients that patients with in the middle or advanced stage of cancer can receive radiotherapy and/or chemotherapy normally under protective measures.


title: The impact of the SARS epidemic on the utilization of medical services: SARS and the fear of SARS.

text: Using interrupted time-series analysis and National Health Insurance data between January 2000 and August 2003, this study assessed the impacts of the severe acute respiratory syndrome (SARS) epidemic on medical service utilization in Taiwan. At the peak of the SARS epidemic, significant reductions in ambulatory care (23.9%), inpatient care (35.2%), and dental care (16.7%) were observed. People's fears of SARS appear to have had strong impacts on access to care. Adverse health outcomes resulting from accessibility barriers posed by the fear of SARS should not be overlooked.


Cryptosporidiosis and proliferative ileitis in a hamster.


title: Inhibitory effects of recombinant human cystatin C on human coronaviruses.

text: Cystatin C, a potent inhibitor of cysteine proteases such as papain and cathepsin B, was examined for its effect on human coronaviruses OC43 and 229e. Both viruses were greater than 99% inhibited by 0.1 mM inhibitor. Endpoint titrations showed that inhibiting activity paralleled that of leupeptin, a serine and cysteine protease inhibitor, and indicated that 1 to 2 microM inhibitor, slightly above physiologic levels, was effective.


Q & A: Bob Michell.


title: Viral antibody studies of laboratory dogs with diarrheal disease.

text: Viral antibody studies were done on laboratory dogs in an epizootic of gastrointestinal disease. Increased hemagglutination-inhibition antibody titers to a parvovirus (PV) antigenically related to feline panleukopenia virus were found in convalescent serum specimens of 78% (20/26) of the affected dogs and in 83% (5/6) of apparently healthy dogs. With one exception, all dogs tested had significant levels of hemagglutination-inhibition antibody to this PV. Similar increased antibody titers were found to feline panleukopenia virus. Also, neutralizing antibody responses were detected to the canine coronavirus in 24% (6/25) and canine herpesvirus in 45% (10/22) of the affected dogs. However, antibody titers did not increase to canine distemper virus, infectious canine hepatitis virus, canine parainfluenza virus, or minute virus of canines. Subsequent serotesting of the colony provided evidence that additional PV infections occurred in pups from each of 8 litters born 3 to 8 months after the epizootic. These findings indicated the continued presence of the PV for more than 1 year in the infected colony. Of 19 laboratory personnel who worked with the affected dogs, none, including 4 with a concurrent diarrheal disease, developed or had antibodies to the PV or canine coronavirus.


title: Endovascular stent placement for chronic post-thrombotic symptomatic ilio-femoral venous obstructive lesions: a single-center study of safety, efficacy and quality-of-life improvement.

text: BACKGROUND Post-thrombotic syndrome (PTS) is a frequent complication of deep vein thrombosis (DVT) despite adequate treatment. Venous angioplasty and stent placement has been progressively used to restore and maintain venous patency in PTS patients. This study reports our single-center experience with the use of endovascular treatment for chronic post-thrombotic symptomatic ilio-femoral venous obstructive lesions. METHODS A prospective mono-centric observational cohort study of PTS patients with chronic symptomatic ilio-femoral venous obstructive lesions referred for endovascular treatment was conducted from March 2012 to April 2016. Procedure consisted in recanalization, pre-dilation and self-expandable stenting of stenotic or occluded iliac and/or femoral veins. Severity of PTS, quality-of-life and treatment outcomes were assessed using Villalta scale and Chronic Venous Insufficiency Questionnaire (CIVIQ-20) at baseline and 3 months after the procedure. Imaging follow-up was based on duplex ultrasound (US) and computed tomography (CT). RESULTS Twenty-one patients (11 females, 10 males; median age, 41 years; range, 32-60) were included. Recanalization and stenting was successfully accomplished in all prime procedures, 4 patients benefitted from an additional procedure. Immediate technical success rate was 96% considering 25 procedures, performed without any complications. Median follow-up was 18 months (range, 6-30 months) with a 90.5% stent patency rate. Villalta score significantly decreased from baseline compared with 3 months after the procedure [14 (range, 11-22) and 5 (range, 1-10), respectively, P<0.0001], showing a significant decrease in the severity of PTS. CIVIQ-20 score significantly decreased from baseline compared with 3 months after stenting [48.5 (range, 39-73) and 26.5 (range, 21-45), respectively, P<0.0001] thus showing a significant improvement of quality-of-life. Post-procedural CIVIQ-20 score was significantly associated with Villalta score (95% CI, 1.53-2.95; P<0.0001). CONCLUSIONS Our results confirm the high clinical success rate and safety of endovascular PTS treatment and highlight the significant impact of stenting on the quality of life of patients with chronic symptomatic ilio-femoral venous obstructive lesions.


title: Metabolic and Hormonal Determinants of Glomerular Filtration Rate and Renal Hemodynamics in Severely Obese Individuals.

text: OBJECTIVE Renal function is often compromised in severe obesity. A true measurement of glomerular filtration rate (GFR) is unusual, and how estimation formulae (EstForm) perform in such individuals is unclear. We characterized renal function and hemodynamics in severely obese individuals, assessing the reliability of EstForm. METHODS We measured GFR (mGFR) by iohexol plasma clearance, renal plasma flow (RPF) by 123I-ortho-iodo-hippurate, basal and stimulated vascular renal indices, endothelium-dependent and -independent vasodilation using flow-mediated dilation (FMD) as well as metabolic and hormonal profile in morbid, otherwise healthy, obese subjects. RESULTS Compared with mGFR, the better performing EstForm was CKD-EPI (5.3 ml/min/1.73 m2 bias by Bland-Altman analysis). mGFR was directly related with RPF, total and incremental glucose AUC, and inversely with PTH and h8 cortisol. Patients with mGFR below the median shown significantly higher PTH and lower vitamin D3. Basal or dynamic renal resistive index, FMD, pulse wave velocity were not related with mGFR. In an adjusted regression model, renal diameter and plasma flow remained related with mGFR (R2 = 0.67), accounting for 15% and 21% of mGFR variance, respectively. CONCLUSIONS CKD-EPI formula should be preferred in morbid obesity; glucose increments during oral glucose tolerance test correlate with hyperfiltration; RPF and diameter are independent determinants of mGFR; slightly high PTH values, frequent in obesity, might influence mGFR.


title: Progressive and nonresolving pneumonia.

text: PURPOSE OF REVIEW Patients with progressive and/or nonresolving community-acquired pneumonia are at risk for increased morbidity and mortality. It is critical to be able to identify patients at risk to institute early appropriate therapy. The purpose of this review is to summarise the most updated developments in this area. RECENT FINDINGS This review will glean from the recent literature clinical, laboratory, and radiologic findings that help identify patients at risk for such complications of their pneumonia. New studies will be reviewed that have identified some of the causes for treatment failures including the type of pathogen and discordant antimicrobial therapy. It will also discuss newly recognised and emerging infectious diseases that may result in progressive or nonresponding pneumonia including severe acute respiratory syndrome, avian influenzae, severe group A streptococcal disease, and community-acquired methicillin-resistant Staphylococcus aureus. Promising treatments have been identified for patients with progressive pneumonia due to an overwhelming host immune response including activated protein C and intravenous immunoglobulin. SUMMARY Both progressive and nonresolving pneumonia represent treatment failure as a result of inappropriate initial therapy, a noninfectious cause, or an overwhelming immune response. It is critical to be able to identify patients with nonresponding pneumonia and to identify patients at risk for progressive pneumonia to institute appropriate therapy.


Influenza in the Eastern Mediterranean Region: identifying the unknowns for detection and control of epidemic and pandemic threats.


title: Intracellular restriction of a productive noncytopathic coronavirus infection.

text: Virus infection in vitro can either result in a cytopathic effect (CPE) or proceed without visible changes in infected cells (noncytopathic infection). We are interested in understanding the mechanisms controlling the impact of coronavirus infection on host cells. To this end, we compared a productive, noncytopathic infection of murine hepatitis virus (MHV) strain A59 in the fibroblastlike cell line NIH 3T3 with cytopathic MHV infections. Infected NIH 3T3 cells could be cultured for up to 4 weeks without apparent CPE and yet produce virus at 10(7) to 10(8) PFU/ml. Using flow cytometry, we demonstrated that NIH 3T3 cells expressed as much MHV receptor CEACAM1 as other cell lines which die from MHV infection. In contrast, using quantitative reverse transcription-PCR and metabolic labeling of RNA, we found that the rate of viral RNA amplification in NIH 3T3 cells was lower than the rate in cells in which MHV induces a CPE. The rate of cellular RNA synthesis in contact-inhibited confluent NIH 3T3 cells was also lower than in cells permissive to cytopathic MHV infection. However, the induction of cellular RNA synthesis in growing NIH 3T3 cells did not result in an increase of either viral RNA amplification or CPE. Our results suggest that a specific, receptor CEACAM1-independent mechanism restricting coronaviral RNA synthesis and CPE is present in NIH 3T3 and, possibly, other cells with preserved contact inhibition.


title: Comparison of the replication of influenza A viruses in Chinese ring-necked pheasants and chukar partridges.

text: We investigated the replication and transmission of avian influenza A viruses in two species thought to be intermediate hosts in the spread of influenza A viruses in live poultry markets: Chinese ring-necked pheasants and chukar partridges. All 15 hemagglutinin subtypes replicated in pheasants, and most subtypes transmitted to naïve contact pheasants, primarily via the fecal-oral route. Many viruses were shed from the gastrointestinal tract of experimentally inoculated pheasants for 14 days or longer. Virus was isolated from the cloacal swabs of one contact pheasant for an unprecedented 45 days. Chukar partridges were less susceptible to infection with avian influenza viruses. The viruses that replicated in chukar partridges were isolated for 7 days after experimental inoculation, predominantly from the respiratory tract. We detected high neutralizing antibody titers with correspondingly low levels of serum hemagglutination inhibition antibody titers in pheasants and chukar partridges when chicken red blood cells were used in serological analyses. When horse erythrocytes were used, antibody titers were comparable to those obtained by using the neutralization assay. More importantly, the results suggested that pheasants can serve as a reservoir of influenza virus. Because of their continuous asymptomatic infection and longer stay in the markets, pheasants are ideal "carriers" of influenza A viruses. Their continued presence in live markets contributes to the perpetuation and genetic interaction of influenza viruses there. On the basis of our findings, it does not make good sense to ban quail but not pheasants from the live markets.


title: Investigation of an outbreak of acute respiratory disease in côte d'ivoire in april 2007.

text: BACKGROUND This study aim was to investigate an outbreak of human cases of unexplained influenza-like illness and fatal acute respiratory infection (ARI), with simultaneous poultry illness and high mortality raising concerns of possible influenza A (H5N1), virus in Cote d'Ivoire in February and March 2007. MATERIALS AND METHODS To investigate the outbreak, we conducted active surveillance in the community and reviewed health registries. Persons meeting the case definition were asked to provide nasopharyngeal specimens. On the basis of clinical and epidemiological information, specimens were tested using conventional RT-PCR for the M gene of the influenza viruses and hemagglutinin H5 of avian influenza A (H5N1), virus; negative samples were tested for other respiratory viruses. Specimens from healthy animals were also collected. RESULTS Between October 2006, and February 2007, 104 suspected cases of Acute Respiratory Disease that included; 31 deaths recorded. We collected and tested 73 nasopharyngeal specimens; of which, 2, were positive for human Coronavirus OC43 and 1 for influenza C virus. No pathogens were identified in animal specimens. CONCLUSIONS The investigation quickly ruled out influenza A (H5N1), virus as the cause and found laboratory-confirmed cases of influenza C virus and human Coronavirus OC 43 for the first time in both Côte d'Ivoire and in a Sub-Saharan African country. However we were not able to show that these viruses caused the outbreak. Monitoring of influenza viruses must be a priority but other respiratory viruses and non-viral causes may be of interest too.


Combined laparoscopic excisional surgery for synchronous endometrial and rectal adenocarcinoma in an obese woman.


title: The distribution of human coronavirus strain 229E on the surface of human diploid cells.

text: The distribution of human coronavirus strain 229E (HCV 229E) particles on the surface of human diploid (MRCc) cells was examined. Virus particles showed a totally random distribution on fixed cells and on cells to which virus had been adsorbed in the cold. A marked redistribution of virus particles was observed on warming virus-cell preparations to 33 degrees C for 20 min, the peripheral areas of the cell becoming relatively devoid of virus particles while the majority of particles were now located some distance from the edge of the cell. Redistribution did not occur in the presence of metabolic inhibitors.


title: A positive-strand RNA virus with three very different subgenomic RNA promoters.

text: Numerous RNA viruses generate subgenomic mRNAs (sgRNAs) for expression of their 3'-proximal genes. A major step in control of viral gene expression is the regulation of sgRNA synthesis by specific promoter elements. We used barley yellow dwarf virus (BYDV) as a model system to study transcriptional control in a virus with multiple sgRNAs. BYDV generates three sgRNAs during infection. The sgRNA1 promoter has been mapped previously to a 98-nucleotide (nt) region which forms two stem-loop structures. It was determined that sgRNA1 is not required for BYDV RNA replication in oat protoplasts. In this study, we show that neither sgRNA2 nor sgRNA3 is required for BYDV RNA replication. The promoters for sgRNA2 and sgRNA3 synthesis were mapped by using deletion mutagenesis. The minimal sgRNA2 promoter is approximately 143 nt long (nt 4810 to 4952) and is located immediately downstream of the putative sgRNA2 start site (nt 4809). The minimal sgRNA3 core promoter is 44 nt long (nt 5345 to 5388), with most of the sequence located downstream of sgRNA3 start site (nt 5348). For both promoters, additional sequences upstream of the start site enhanced sgRNA promoter activity. These promoters contrast to the sgRNA1 promoter, in which almost all of the promoter is located upstream of the transcription initiation site. Comparison of RNA sequences and computer-predicted secondary structures revealed little or no homology between the three sgRNA promoter elements. Thus, a small RNA virus with multiple sgRNAs can have very different subgenomic promoters, which implies a complex system for promoter recognition and regulation of subgenomic RNA synthesis.


title: An exposed domain in the severe acute respiratory syndrome coronavirus spike protein induces neutralizing antibodies.

text: Exposed epitopes of the spike protein may be recognized by neutralizing antibodies against severe acute respiratory syndrome (SARS) coronavirus (CoV). A protein fragment (S-II) containing predicted epitopes of the spike protein was expressed in Escherichia coli. The properly refolded protein fragment specifically bound to the surface of Vero cells. Monoclonal antibodies raised against this fragment recognized the native spike protein of SARS CoV in both monomeric and trimeric forms. These monoclonal antibodies were capable of blocking S-II attachment to Vero cells and exhibited in vitro antiviral activity. These neutralizing antibodies mapped to epitopes in two peptides, each comprising 20 amino acids. Thus, this region of the spike protein might be a target for generation of therapeutic neutralizing antibodies against SARS CoV and for vaccine development to elicit protective humoral immunity.


title: Identification of linear heparin-binding peptides derived from human respiratory syncytial virus fusion glycoprotein that inhibit infectivity.

text: It has been shown previously that the fusion glycoprotein of human respiratory syncytial virus (RSV-F) interacts with cellular heparan sulfate. Synthetic overlapping peptides derived from the F-protein sequence of RSV subtype A (strain A2) were tested for their ability to bind heparin using heparin-agarose affinity chromatography (HAAC). This evaluation identified 15 peptides representing eight linear heparin-binding domains (HBDs) located within F1 and F2 and spanning the protease cleavage activation site. All peptides bound to Vero and A549 cells, and binding was inhibited by soluble heparins and diminished by either enzymatic treatment to remove cell surface glycosaminoglycans or by treatment with sodium chlorate to decrease cellular sulfation. RSV-F HBD peptides were less likely to bind to glycosaminoglycan-deficient CHO-745 cells than parental CHO-K1 cells that express these molecules. Three RSV-F HBD peptides (F16, F26, and F55) inhibited virus infectivity; two of these peptides (F16 and F55) inhibited binding of virus to Vero cells, while the third (F26) did not. These studies provided evidence that two of the linear HBDs mapped by peptides F16 and F55 may mediate one of the first steps in the attachment of virus to cells while the third, F26, inhibited infectivity at a postattachment step, suggesting that interactions with cell surface glycosaminoglycans may play a role in infectivity of some RSV strains.


title: Characterization of murine coronavirus RNA by hybridization with virus-specific cDNA probes.

text: Genome RNA of mouse hepatitis virus (MHV) strain A59 has been used as a template to synthesize two virus-specific probes: cDNArep, representing the majority of sequences of the genome RNA and cDNA3', representing the 3' end of the genome RNA. Molecular hybridization with these cDNAs was used to characterize both genome RNA and intracellular virus-specific RNAs. Hybridization of genome RNAs of MHV strains A59, JHM, and MHV-3 with A59 cDNArep showed that, although these three strains exhibit different pathogenicities, they contain closely related nucleotide sequences. Hybridization of intracellular RNA from MHV-infected cells with virus-specific cDNA shows that (i) the majority of virus-specific RNA is polyadenylated, (ii) virus-specific intracellular RNA contains six subgenomic species of the same polarity as genome RNA and (iii) all subgenomic RNAs contain the same 3' sequences as the genome RNA and thus form a nested set of RNAs.


Detection of body temperature with infrared thermography: accuracy in detection of fever.


The open university.


Characterization of a small plaque mutant of the A59 strain of mouse hepatitis virus defective in cell fusion.


title: The treatment of hospital wastewater: an appraisal.

text: Hospitals discharge considerable amounts of chemicals and microbial agents in their wastewaters. Problem chemicals present in hospital wastewater belong to different groups, such as antibiotics, X-ray contrast agents, disinfectants and pharmaceuticals. Many of these chemical compounds resist normal wastewater treatment. They end up in surface waters where they can influence the aquatic ecosystem and interfere with the food chain. Humans are particularly exposed by the drinking water, produced from surface water. Microbial agents of special concern are multiresistant microbial strains. The latter are suspected to contribute to the spread of antibiotic resistance. In this paper, we will discuss the different approaches towards hospital wastewater treatment. The principle of uncoupling hospitals from public sewers warrants indepth evaluation by technologists and ecotoxicologists as well as public health specialists.


title: Japanese Encephalitis Vaccines.

text: Japanese encephalitis (JE) is a significant human health concern in Asia, Indonesia and parts of Australia with more than 3 billion people potentially at risk of infection with Japanese encephalitis virus (JEV), the causative agent of JE. Given the risk to human health and the theoretical potential for JEV use as a bioweapon, the development of safe and effective vaccines to prevent JEV infection is vital for preserving human health. The development of vaccines for JE began in the 1940s with formalin-inactivated mouse brain-derived vaccines. These vaccines have been shown to induce a protective immune response and to be very effective. Mouse brain-derived vaccines were still in use until May 2011 when the last lots of the BIKEN(®) JE-VAX(®) expired. Development of modern JE vaccines utilizes cell culture-derived viruses and improvements in manufacturing processes as well as removal of potential allergens or toxins have significantly improved vaccine safety. China has developed a live-attenuated vaccine that has proven to induce protective immunity following a single inoculation. In addition, a chimeric vaccine virus incorporating the prM and E structural proteins derived from the live-attenuated JE vaccine into the live-attenuated yellow fever 17D vaccine virus backbone is currently in clinical trials. In this article, we provide a summary of JE vaccine development and on-going clinical trials. We also discuss the potential risk of JEV as a bioweapon with a focus on virus sustainability if used as a weapon.


title: Characterization of an antigenically distinct porcine rotavirus.

text: A porcine virus with rotavirus morphology, which was antigenically unrelated to previously described rotaviruses, is described. Particles with an outer capsid layer measured 75 nm and those lacking the outer layer were 63 nm in diameter. Particles which resembled cores were also identified. The virus was shown to be antigenically distinct from other rotaviruses as judged by immunofluorescence and immune electron microscopy, and it failed to protect piglets from challenge with porcine rotavirus. Analysis of the genome indicated that it was composed of 11 segments of double-stranded RNA with the same overall size range as other rotaviruses but with an unusual segment pattern.


title: Options in topical therapies in the management of patients with acute pain.

text: The traditional cornerstones of analgesic therapy for patients with acute pain have been oral therapies; however, all oral agents exhibit a variety of potentially dose-limiting or intolerable adverse effects in patients. Elderly patients and those with concomitant conditions already being managed with multiple systemic drugs may be particularly susceptible to systemic toxicities with oral analgesic therapies. Topical agents offer an alternative to oral modalities and can effectively treat patients with acute pain while offering lower systemic absorption and conferring little risk of systemic toxicity. The objective of this article is to review the therapeutic usefulness of available topical therapies in their most thoroughly investigated applications, the treatment of patients with acute musculoskeletal and herpetic pain. For example, although heating pads/wraps and cold packs are widely used to alleviate pain associated with sprains, strains, and contusions, evidence of the effectiveness of these methods is lacking. However, there are sufficient data supporting the use of various topical formulations of nonsteroidal anti-inflammatory drugs (NSAIDs) for these indications (ketoprofen gel or patch, ibuprofen gel or cream, and diclofenac gel or patch), and demonstrating markedly less patient risk of systemic toxicity than is associated with oral NSAID therapy. A ketoprofen patch was shown to be effective and well tolerated in the treatment of patients with tendinopathies. In the treatment of acute neck or low back pain, cold and heat therapies have demonstrated limited effectiveness for patients, and the efficacy of topical NSAIDs has not been established. Use of topical NSAID therapy has been useful in reducing acute-phase herpes zoster pain, and the lidocaine 5% patch has been shown to reduce acute herpetic pain intensity once lesions have healed (the patch cannot be applied to open skin lesions). Topical analgesics represent an alternative treatment modality for patients experiencing acute pain who cannot or choose not to take oral therapies.


title: Licking latency with licorice.

text: Numerous viruses cause latent infections in humans, and reactivation often results in pain and suffering. While vaccines for several of these viruses are available or currently being studied in clinical trials, and antiviral therapies have been successful in preventing or treating active infection, therapy to eradicate latent infection has lagged behind. A new study reported in this issue of the JCI shows that treatment of cells latently infected with Kaposi sarcoma-associated herpesvirus (KSHV) with glycyrrhizic acid, a component of licorice, reduces synthesis of a viral latency protein and induces apoptosis of infected cells. This finding suggests a novel way to interrupt latency.


Detection of feline infectious peritonitis virus-like antigen in ferrets.


title: [Virological and serological characteristics of outbreaks and sporadic cases of acute gastroenteritis].

text: Specimens from patients with gastroenteritis (GE) collected during outbreaks and from sporadic cases reported in the USSR in 1979-1984 were examined by electron microscopy (EM), enzyme immunoassay, rotavirus neutralization test in cell culture. All the winter-spring outbreaks and a considerable number (34.9%) of sporadic GE cases were caused by rotaviruses. The summer-autumn outbreaks were of non-rotavirus nature. In water-borne winter-spring outbreaks in adults, severe forms of GE with signs of dehydration were observed. Among infants, cases of virus-carrier state were detected. The rate of rotavirus detection by EM in winter-spring outbreaks depended on the time of specimen collection and decreased after 4 days from the onset of the disease. Apart from rotaviruses, adeno-, astro-, calici-, coronaviruses, and picornavirus-like particles were detected by EM in feces from GE patients.


title: Detection and molecular characterization of gene 3 and 5 of turkey coronavirus from turkeys with severe enteritis in Brazil.

text: Turkey coronavirus (TCoV) is a causative agent associated with poult enteritis and mortality syndrome (PEMS) in turkeys worldwide. The disease is an acute, highly contagious enteric disease that is characterized by depression, anorexia, diarrhea, and high mortality in commercial turkey flocks. The presence of TCoV in 12 intestinal-content samples, from turkey flocks aged between 10 and 104 days and exhibiting severe enteritis, was monitored during the period of 2004 to 2006. TCoV detection was accomplished by a reverse transcriptase-polymerase chain reaction (RT-PCR) through amplification of the 3' UTR region, followed by amplification of genes 3 and 5. Molecular characterization of the viruses was done through amplification of genes 3 and 5 and showed evidence of genetic similarity between them, although they differed from sequences of other TCoVs described in the literature. In relation to gene 3, samples showed a greater relationship with chicken infectious bronchitis virus (IBV), while gene 5 showed greater identity with pheasant coronavirus (PhCoV). Our results suggest that the strategy of amplification of the 3' UTR region, followed by sequencing of genes 3 and 5, has proven to be an effective means of detecting TCoV in intestinal contents.


title: DNA accelerates the inhibition of human cathepsin V by serpins.

text: A balance between proteolytic activity and protease inhibition is crucial to the appropriate function of many biological processes. There is mounting evidence for the presence of both papain-like cysteine proteases and serpins with a corresponding inhibitory activity in the nucleus. Well characterized examples of cofactors fine tuning serpin activity in the extracellular milieu are known, but such modulation has not been studied for protease-serpin interactions within the cell. Accordingly, we present an investigation into the effect of a DNA-rich environment on the interaction between model serpins (MENT and SCCA-1), cysteine proteases (human cathepsin V and human cathepsin L), and cystatin A. DNA was indeed found to accelerate the rate at which MENT inhibited cathepsin V, a human orthologue of mammalian cathepsin L, up to 50-fold, but unexpectedly this effect was primarily effected via the protease and secondarily by the recruitment of the DNA as a "template" onto which cathepsin V and MENT are bound. Notably, the protease-mediated effect was found to correspond both with an altered substrate turnover and a conformational change within the protease. Consistent with this, cystatin inhibition, which relies on occlusion of the active site rather than the substrate-like behavior of serpins, was unaltered by DNA. This represents the first example of modulation of serpin inhibition of cysteine proteases by a co-factor and reveals a mechanism for differential regulation of cathepsin proteolytic activity in a DNA-rich environment.


title: Total sialic acid: an acute phase reactant in cats with a possible role in feline coronavirus infection.

text: The aims of this study were to validate a colorimetric method to measure total sialic acid (TSA) in feline serum and to investigate the serum concentration of TSA in clinically healthy cats seronegative (n = 9) and seropositive (n = 48) for feline coronavirus (FCoV), and in cats affected by feline infectious peritonitis (FIP, n = 28), tumors (n = 20), or inflammation (n = 16). The correlation between TSA and alpha(1)-acid glycoprotein (AGP) was also investigated. The method employed in this study is precise and accurate at TSA levels (in mg/L) commonly encountered in feline serum. No significant differences between seropositive (385.6 +/- 192.2 mg/L) and seronegative (433.5 +/- 179.0 mg/L) cats were detectable, suggesting that the simple infection by FCoVs does not influence TSA levels. Compared with seropositive controls, the concentration of TSA was higher in cats with FIP (556.7 +/- 268.3 mg/L, P = 0.003), tumors (522.5 +/- 294.4 mg/L, P = 0.028), and inflammation (546.8 +/- 208.3 mg/L, P = 0.018). The discriminating power of TSA for FIP is moderate (area under the ROC curve = 0.65) and the likelihood ratio is higher than 3.0 only at high TSA levels. Consequently, TSA could support a diagnosis of FIP only at extremely high serum concentration (> 800 mg/L) or when the pre-test probability of FIP is high. No correlations were found between the TSA and AGP concentrations in cats with FIP, suggesting that sialylated proteins other than AGP are present. Both the antibody titre and the degree of AGP sialylation were negatively correlated with TSA levels, suggesting that increased TSA may contribute to reduce the burden of FCoVs.


title: Novel double-stapling technique for distal oesophageal resection and oesophago-jejunal anastomosis.

text: AIM The restoration of the digestive tract by performing an esophago-jejunal anastomosis (EJA) is a crucial step of the total gastric and distal esophagus surgical resection for esophago-gastric junction (EGJ) cancer. We have already ideated and tested on a cadaver model an innovative technique which could be useful to minimize the risk of complications related to the phase of securing the anvil of the circular stapler prior to perform the EJA. This surgical technique was derived from the well-known "double-stapling Knight and Griffen" one that was described for the rectal resection. We used the following described technique in 20 patients with EGJ cancer and it is efficient, reliable, safe, easy to learn and easy to perform. MATERIALS AND METHODS From August 2014 to May 2015, 20 patients (14 male and 6 female) underwent surgery for esophagogastric junction cancer: In all patients a distal esophageal resection and total gastrectomy was performed. Through the trans-hiatal access, the free margins of the esophageal stump were suspended and the anvil of a circular stapler on a new dedicated and registered support bar was inserted into the lumen. Subsequently, the linear suturing stapler is closed over the bar and then fired to suture the distal stump of the esophagus; after the confirmation of a negative margin, the bar is retracted and the push-rod of the anvil is pulled out through the linear suture. Finally, the anastomosis is performed with the classic technique by using a circular stapler. RESULTS No postoperative mortality occurred; postoperative course has been uneventful for 18 patients. One patient developed anastomotic fistula that has been treated conservatively with endoscopic prothesis, removed after 20 days. One patient developed in 3 POD myocardial infarction Mean Hospital stay has been 14 days (range 7-20 days). CONCLUSIONS The aim of our new procedure is the insertion the anvil of a common circular stapler without handsewn securing; this is to reduce the technical difficulties related to the hand-sewn securing into a deep and narrow anatomic location, typical of the trans-hiatal approach. KEY WORDS Anastomosis, Oesophago-gastric junction cancer, Stapler, Trans-hiatal.


title: Novel beta-barrel fold in the nuclear magnetic resonance structure of the replicase nonstructural protein 1 from the severe acute respiratory syndrome coronavirus.

text: The nonstructural protein 1 (nsp1) of the severe acute respiratory syndrome coronavirus has 179 residues and is the N-terminal cleavage product of the viral replicase polyprotein that mediates RNA replication and processing. The specific function of nsp1 is not known. Here we report the nuclear magnetic resonance structure of the nsp1 segment from residue 13 to 128, which represents a novel alpha/beta-fold formed by a mixed parallel/antiparallel six-stranded beta-barrel, an alpha-helix covering one opening of the barrel, and a 3(10)-helix alongside the barrel. We further characterized the full-length 179-residue protein and show that the polypeptide segments of residues 1 to 12 and 129 to 179 are flexibly disordered. The structure is analyzed in a search for possible correlations with the recently reported activity of nsp1 in the degradation of mRNA.


title: Using protein homology models for structure-based studies: approaches to model refinement.

text: Homology modeling is a computational methodology to assign a 3-D structure to a target protein when experimental data are not available. The methodology uses another protein with a known structure that shares some sequence identity with the target as a template. The crudest approach is to thread the target protein backbone atoms over the backbone atoms of the template protein, but necessary refinement methods are needed to produce realistic models. In this mini-review anchored within the scope of drug design, we show the validity of using homology models of proteins in the discovery of binders for potential therapeutic targets. We also report several different approaches to homology model refinement, going from very simple to the most elaborate. Results show that refinement approaches are system dependent and that more elaborate methodologies do not always correlate with better performances from built homology models.


title: [Simultaneous detection of respiratory viruses and influenza A virus subtypes using multiplex PCR].

text: This study was conducted to investigate the respiratory viruses and subtyping of influenza A virus when positive by multiplex PCR in patients with flu-like symptoms, after the pandemic caused by influenza A (H1N1)pdm09. Nasopharyngeal swab samples collected from 700 patients (313 female, 387 male; age range: 24 days-94 yrs, median age: 1 yr) between December 2010 - January 2013 with flu-like symptoms including fever, headache, sore throat, rhinitis, cough, myalgia as defined by the World Health Organization were included in the study. Nucleic acid extractions (Viral DNA/RNA Extraction Kit, iNtRON, South Korea) and cDNA synthesis (RevertAid First Strand cDNA Synthesis Kits, Fermentas, USA) were performed according to the manufacturer's protocol. Multiplex amplification of nucleic acids was performed using DPO (dual priming oligonucleotide) primers and RV5 ACE Screening Kit (Seegene, South Korea) in terms of the presence of influenza A (INF-A) virus, influenza B (INF-B) virus, respiratory syncytial virus (RSV), and the other respiratory viruses. PCR products were detected by automated polyacrylamide gel electrophoresis using Screen Tape multiple detection system. Specimens which were positive for viral nucleic acids have been further studied by using specific DPO primers, FluA ACE Subtyping and RV15 Screening (Seegene, South Korea) kits. Four INF-A virus subtypes [human H1 (hH1), human H3 (hH3), swine H1 (sH1), avian H5 (aH5)] and 11 other respiratory viruses [Adenovirus, parainfluenza virus (PIV) types 1-4, human bocavirus (HBoV), human metapneumovirus (HMPV), rhinovirus types A and B, human coronaviruses (HCoV) OC43, 229E/NL63] were investigated with those tests. In the study, 53.6% (375/700) of the patients were found to be infected with at least one virus and multiple respiratory virus infections were detected in 15.7% (59/375) of the positive cases, which were mostly (49/59, 83%) in pediatric patients. RSV and rhinovirus coinfections were the most prevalent (18/29, 62.7%) dual infections. The distribution of 436 respiratory viruses identified from 375 patients were as follows; 189 (43.3%) RSV, 93 (21.4%) rhinovirus, 86 (19.8%) INF-A, seven (1.6%) INF-B, 22 (5%) PIV types 1-3, 14 (3.2%) HMPV, 11 (2.5%) HCoV, nine (2%) HBoV, and five (1.2%) adenovirus. Fifty-five (64%) out of 86 INF-A viruses were subtyped as hH3, 24 (27.9%) were sH1 and seven (8.1%) were hH1. Avian H5 was not detected in any samples. The overall prevalence rates of INF-A, INF-B, RSV and other respiratory viruses were 12%, 1%, 27%, and 14.6%, respectively. RSV was the most prevalent respiratory agent in pediatric (161/313, 51%) cases, while INF-A virus in adult (24/62, 38.7%) patients. Influenza viruses were detected as responsible pathogens in 13.3% (93/700) of the patients with flu-like symptoms. Among the cases, a 1-month-old baby was infected with three virus strains (INF-A hH1+INF-A sH1+HCoV OC43) and a 82-year-old patient was infected with two INF-A virus subtypes (hH3 + sH1). INF-A viruses were mostly detected (79/86) in winter period, from December to March. INF-A virus sH1, was the most prevalent subtype in flu cases till February 2011 (22/86), after replaced by INF-A virus hH3. Beginning from February 2012, a significant increase observed in the cases infected with INF-A virus subtype hH3 (39/86). In conclusion, the identification and surveillance of influenza virus types and subtypes circulating in populations have importance both for epidemiological data and selection of vaccine strains.


title: Development and comparison of the real-time amplification based methods--NASBA-Beacon, RT-PCR taqman and RT-PCR hybridization probe assays--for the qualitative detection of sars coronavirus.

text: The aim of this study was to develop a rapid, sensitive and robust procedure for the qualitative detection of SARS coronavirus RNA. Three unique detection formats were developed for real-time RNA amplification assays: a post amplification detection step with a virus-specific internal capture probe based on Taqman (RT-PCR TaqMan assay), hybridization probe (RT-PCR hybridization probe assay) and a real-time assay with virus-specific molecular beacon probes (NASBA-Beacon assay). The analytical sensitivity or reproducibility of the test results among those three assays was compared. All assays yielded results by detecting SARS coronavirus targeting the BNI-1 region in less than 2 hours. RNA detection by all the formats was unaffected by the presence of human sputum. The limits of detection were at least 10 copies of input RNA for both RT-PCR formats (RT-PCR TaqMan and RT-PCR hybridization probe assays), while the NASBA-Beacon assay could detect as little as 1 copy per reaction, with high reproducibility of the coefficient of variation (CV) of <10. These results demonstrate that real-time NASBA provides a rapid and sensitive alternative to RT-PCR for the routine qualitative assay of sputum for SARS corona viral RNA detection.


Differential diagnostic blocking ELISA for use in conjunction with modified-live IBRV gIII-deleted marker vaccine.


title: 2-D graphical representation of protein sequences and its application to coronavirus phylogeny.

text: Based on a five-letter model of the 20 amino acids, we propose a new 2-D graphical representation of protein sequence. Then we transform the 2-D graphical representation into a numerical characterization that will facilitate quantitative comparisons of protein sequences. As an application, we construct the phylogenetic tree of 56 coronavirus spike proteins. The resulting tree agrees well with the established taxonomic groups.


title: Nuclear localization of non-structural protein 1 and nucleocapsid protein of equine arteritis virus.

text: RNA synthesis (genome replication and subgenomic mRNA transcription) directed by equine arteritis virus (EAV; family Arteriviridae, order Nidovirales) occurs on modified cytoplasmic membranes to which most viral replicase subunits localize. Remarkably, a fraction of non-structural protein 1 (nsp1), a protein essential for transcription but dispensable for genome replication, is present in the host cell nucleus, in particular during the earlier stages of infection. Expression of GFP-tagged fusion proteins revealed that nsp1 is actively imported into the nucleus. Although the signals responsible for nsp1 transport could not be identified, our studies revealed that another EAV protein with a partially nuclear localization, the nucleocapsid (N) protein, utilizes the CRM1-mediated nuclear export pathway. Inactivation of this pathway with the drug leptomycin B resulted in the unexpected and immediate nuclear retention of all N protein molecules, thus revealing that the protein shuttles between cytoplasm and nucleus before playing its role in cytoplasmic virus assembly.


Rotavirus diarrhoea in patient with antibody to human immunodeficiency virus (HIV)


title: Sequence motifs involved in the regulation of discontinuous coronavirus subgenomic RNA synthesis.

text: Coronavirus transcription leads to the synthesis of a nested set of mRNAs with a leader sequence derived from the 5' end of the genome. The mRNAs are produced by a discontinuous transcription in which the leader is linked to the mRNA coding sequences. This process is regulated by transcription-regulating sequences (TRSs) preceding each mRNA, including a highly conserved core sequence (CS) with high identity to sequences present in the virus genome and at the 3' end of the leader (TRS-L). The role of TRSs was analyzed by reverse genetics using a full-length infectious coronavirus cDNA and site-directed mutagenesis of the CS. The canonical CS-B was nonessential for the generation of subgenomic mRNAs (sgmRNAs), but its presence led to transcription levels at least 10(3)-fold higher than those in its absence. The data obtained are compatible with a transcription mechanism including three steps: (i) formation of 5'-3' complexes in the genomic RNA, (ii) base-pairing scanning of the nascent negative RNA strand by the TRS-L, and (iii) template switching during synthesis of the negative strand to complete the negative sgRNA. This template switch takes place after copying the CS sequence and was predicted in silico based on high base-pairing score between the nascent negative RNA strand and the TRS-L and minimum DeltaG.


title: Development of a nested PCR assay for detection of feline infectious peritonitis virus in clinical specimens.

text: A diagnostic test for feline infectious peritonitis virus (FIPV) infection based on a nested PCR (nPCR) assay was developed and tested with FIPV, feline enteric coronavirus (FECV), canine coronavirus (CCV), and transmissible gastroenteritis virus (TGEV) and clinical fluid samples from cats with effusive feline infectious peritonitis (FIP). The target sequence for the assay is in the S1 region of the peplomer protein E2 gene. A vaccine strain of FIPV and two wild-type FIPV strains tested positive, but FECV, TGEV, and CCV tested negative. Preliminary tests with 12 cats with clinical evidence of effusive FIP and 11 cats with an illness associated with effusions, but attributed to other causes, were performed. Eleven of the 12 cats with effusive FIP tested positive, while 1 was negative. Ten of the 11 cats ill from other causes tested negative, while 1 was positive. On the basis of clinical laboratory and histopathologic criteria, the preliminary sensitivity and specificity of the assay were 91.6 and 94%, respectively.


title: Characterisation of a papain-like proteinase domain encoded by ORF1a of the coronavirus IBV and determination of the C-terminal cleavage site of an 87 kDa protein.

text: Our previous studies have shown that two overlapping papain-like proteinase domains (PLPDs) encoded by the IBV sequence from nucleotides 4155 to 5550 is responsible for cleavage of the ORF 1a polyprotein to an 87 kDa protein. In this study, we demonstrate that only the more 5' one of the two domains, PLPD-1 encoded between nucleotides 4155 and 5031, is required for processing to the 87 kDa protein. Site-directed mutagenesis studies have shown that the Cys1274 and His1435 residues are essential for the PLPD-1 activity, suggesting that they may be the components of the catalytic centre of this proteinase. Coexpression and immunoprecipitation studies have further revealed that PLPD can interact with the 87 kDa protein. Meanwhile, data obtained from the construction and expression of a series of deletion mutants have indicated that the 87 kDa protein is encoded by the 5'-most 2600 bp part of ORF1a. further deletion and mutagenesis studies are underway to determine precisely the C-terminal cleavage site of the 87 kDa protein.


title: Lung damage may induce thrombocytopenia.

text: Thrombocytopenia is common in patients with lung diseases. Lung injury is associated with deranged platelet homeostasis. The potential mechanisms underlying this association were investigated in this study. The number and morphology of circulating megakaryocytes (MK) were investigated in vivo during cardiopulmonary bypass (CPB) in patients undergoing elective cardiac surgery for heart disease. The association between lung injury and thrombocytopenia was further investigated in vitro in rats with lung injury. MK and platelet counts, and platelet activation between the prepulmonary (right atrial) and postpulmonary (left atrial) blood were also compared. In human, the total number of MK in central venous was higher than those of peripheral arteries during normal circulation. During CPB, the total MK and Type-4 MK of central venous and peripheral arteries were significant increased when compared with that in normal circulation. In healthy rats, the postpulmonary blood had lower MK count, higher platelet count, and similar P-selectin expression. However, the lung-damaged animals showed no such differences in either MK or platelet count, but P-selectin expression was greater in the postpulmonary blood. Peripheral platelets in the lung-injured rats were significantly lower than those in their respective controls. The lungs may play an active role in the regulation of MK levels and the lung damage may reduce circulating platelets.


title: [Immunohistochemical investigation of lungs with severe acute respiratory syndrome].

text: OBJECTIVE To investigate the roles of different cells in the pulmonary lesions in the severe acute respiratory syndrome (SARS) patients. METHODS The monoclonal antibodies of CD8, CD20, CD34, LCA, CD56, CD68, and AE1/AE3 are used to demonstrate the different cells in the lung specimens of SARS patients in order to study the patterns of cell responses in this new disease. Meanwhile the HE stained slides were also carefully studied to compare with the results of immunohistochemical staining. RESULTS The number of capillaries increased and the capillaries clearly outlined the contour of alveolar wall from beginning to early stage of organization, the number of lymphocytes decreased sharply while the number of macrophage remarkably increased, together with proliferation of type II pneumocytes. The numbers of blood vessels decreased in the fibrotic and consolidated lung tissue, and the vessel cavities enlarged, losing the normal contour of alveolar septa. CONCLUSIONS The lesions in the lung from SARS patients are consisted of the tissue reaction to the inflammatory injury, including extensive exudation, capillary proliferation, fibrosis, and obvious infiltration of macrophages which may play a key role in the pathogenesis of pulmonary lesions of SARS.


title: Single incision laparoscopic cholecystectomy.

text: BACKGROUND As surgeons embrace the concept of increasingly less invasive surgery, techniques using only a single small incision have begun to gain traction. Multiple case series managed by a single-incision laparoscopic cholecystectomy (SILC) have been published. The objective of this study is to describe the short-term outcomes of SILC in our institution. METHODS A retrospective review was done for 51 patients who underwent SILC between 2009-2012. The operative time, surgical technique, conversion rate, and postoperative complications were reported. RESULTS SILC was performed for 51 patients, all women, with a mean age of 35.6 years (range=19-62). Their body mass index(BMI) ranged from 16.8-35.6 kg m2 with a mean of 20.4. Twelve patients (23.52 %) had acute cholecystitis. The mean operating time was 58.6 minutes (range=45-95 min). Incidents were encountered in 6 patients (11.76%) and were related to intraoperative bleeding. There was a single conversion to the standard laparoscopic procedure (1.9%) and in 5 cases an accessory needle grasp was used (9.8%). Two patients (3.9%)presented postoperative complications (wound infections) and the mean hospital stay was 1.6 days. CONCLUSIONS SILC is feasible and provides a promising alternative to natural orifice transluminal endoscopic surgery for scarless laparoscopic cholecystectomy. However, routine application of this novel technique requires an evaluation of its safety and cost effectiveness in larger studies.


title: Pasteurella multocida infections in rockhopper penguins (Eudyptes chrysocome) from Campbell Island, New Zealand.

text: During an investigation into the population decline of rockhopper penguins (Eudyptes chrysocome) on Campbell Island, New Zealand, avian cholera (Pasteurella multocida) was found in dead adults and chicks. An RNA enveloped virus was isolated from Ixodes uriae, a tick which commonly parasitizes rockhopper penguins on the island. It is not known whether this virus is virulent for penguins. No evidence was obtained to suggest that avian cholera was the principal cause for the decline in the rockhopper penguin population.


title: Mucosal immunization of calves with recombinant bovine adenovirus-3: induction of protective immunity to bovine herpesvirus-1.

text: To determine the potential of replication-competent (E3-deleted) bovine adenovirus-3 (BAV-3) as a delivery system for vaccine antigens in calves, we evaluated the ability of recombinant BAV-3 expressing different forms of of bovine herpesvirus-1 (BHV-1) glycoprotein gD to protect against BHV-1 infection in calves that had pre-existing BAV-3 specific antibodies. Three- to four-month-old calves, vaccinated intranasally with recombinant BAV-3 expressing full-length gD (BAV3.E3gD) or a truncated version of gD (gDt) (BAV3.E3gDt), or with E3-deleted BAV-3 (BAV3.E3d; control), were challenged with BHV-1 strain 108. Vaccination with BAV3.E3gD or BAV3.E3gDt induced gD-specific antibody responses in serum and nasal secretions, and primed calves for gD-specific lymphoproliferative responses. In addition, all calves developed complement-independent neutralizing antibodies against BHV-1. Protection against viral challenge was observed in calves vaccinated with recombinant BAV3.E3gD or BAV3.E3gDt as shown by a significant reduction in body temperature and clinical disease, and a partial reduction in the amount and duration of virus excretion in nasal secretions. These results indicate that replication-competent BAV-3-based vectors can induce protective immune responses in calves (the natural host) that have pre-existing BAV-3-specific antibodies.


title: Detection of in ovo-inoculated infectious bronchitis virus by immunohistochemistry and in situ hybridization with a riboprobe in epithelial cells of the lung and cloacal bursa.

text: In situ hybridization and immunohistochemistry were utilized to identify tissues infected in ovo with infectious bronchitis virus (IBV). Chicken embryos were inoculated in ovo (chorioallantoic sac) with the Arkansas (Ark) serotype of IBV at 18 days of age. At 24, 48, 72, and 120 hr postinfection (HPI), bursa, lung, spleen, heart, and thymus were collected, fixed in 10% neutral buffered formalin, and paraffin embedded. The digoxigenin-labeled antisense S1 riboprobe detected viral mRNA in the cytoplasm of respiratory epithelial cells in the primary bronchus at 24, 48, and 72 HPI. Viral mRNA was detected in bursa samples collected at 48 hr. Immunohistochemistry detected viral antigens in epithelial cells of the parabronchi and bursal tissues at 24 and 48 hr, respectively. No viral mRNA or antigen was detected by in situ hybridization or immunohistochemistry, respectively, in heart, thymus, or spleen at any time after inoculation. On the basis of these data, IBV apparently initially infects lung tissue, then migrates to and infects cells of the bursa. These results indicate that in situ hybridization can be useful in detection of IBV-infected chickens and in understanding the pathogenesis and virulence of IBV infection.


Suicide prevention and intervention: a call to all doctors.


Authors reply: Middle East respiratory syndrome coronavirus (MERS-CoV) in dromedary camels: are dromedary camels a reservoir for MERS-CoV?


The avian coronavirus multiplication strategy.


title: Uses of inorganic hypochlorite (bleach) in health-care facilities.

text: Hypochlorite has been used as a disinfectant for more than 100 years. It has many of the properties of an ideal disinfectant, including a broad antimicrobial activity, rapid bactericidal action, reasonable persistence in treated potable water, ease of use, solubility in water, relative stability, relative nontoxicity at use concentrations, no poisonous residuals, no color, no staining, and low cost. The active species is undissociated hypochlorous acid (HOCl). Hypochlorites are lethal to most microbes, although viruses and vegetative bacteria are more susceptible than endospore-forming bacteria, fungi, and protozoa. Activity is reduced by the presence of heavy metal ions, a biofilm, organic material, low temperature, low pH, or UV radiation. Clinical uses in health-care facilities include hyperchlorination of potable water to prevent Legionella colonization, chlorination of water distribution systems used in hemodialysis centers, cleaning of environmental surfaces, disinfection of laundry, local use to decontaminate blood spills, disinfection of equipment, decontamination of medical waste prior to disposal, and dental therapy. Despite the increasing availability of other disinfectants, hypochlorites continue to find wide use in hospitals.


title: Cardiovascular complications of severe acute respiratory syndrome.

text: BACKGROUND AND AIMS Severe acute respiratory syndrome (SARS) is a virulent viral infection that affects a number of organs and systems. This study examined if SARS may result in cardiovascular complications. METHODS AND RESULTS 121 patients (37.5 (SD13.2) years, 36% male) diagnosed to have SARS were assessed continuously for blood pressure, pulse, and temperature during their stay in hospital. Hypotension occurred in 61 (50.4%) patients in hospital, and was found in 28.1%, 21.5%, and 14.8% of patients during the first, second, and third week, respectively. Only one patient who had transient echocardiographic evidence of impaired left ventricular systolic function required temporary inotropic support. Tachycardia was present in 87 (71.9%) patients, and was found in 62.8%, 45.4%, and 35.5% of patients from the first to third week. It occurred independent of hypotension, and could not be explained by the presence of fever. Tachycardia was also present in 38.8% of patients at follow up. Bradycardia only occurred in 18 (14.9%) patients as a transient event. Reversible cardiomegaly was reported in 13 (10.7%) patients, but without clinical evidence of heart failure. Transient atrial fibrillation was present in one patient. Corticosteroid therapy was weakly associated with tachycardia during the second (chi(2) = 3.99, p = 0.046) and third week (chi(2) = 6.53, p = 0.01), although it could not explain tachycardia during follow up. CONCLUSIONS In patients with SARS, cardiovascular complications including hypotension and tachycardia were common but usually self limiting. Bradycardia and cardiomegaly were less common, while cardiac arrhythmia was rare. However, only tachycardia persisted even when corticosteroid therapy was withdrawn.


title: Unraveling the complexities of the interferon response during SARS-CoV infection.

text: Viruses employ different strategies to circumvent the antiviral actions of the innate immune response. SARS coronavirus (SARS-CoV), a virus that causes severe lung damage, encodes an array of proteins able to inhibit induction and signaling of type-I interferons. However, recent studies have demonstrated that interferons are produced during SARS-CoV infection in humans and macaques. Furthermore, nuclear translocation of activated STAT1 and a range of interferon-stimulated genes could be demonstrated in the lungs of SARS-CoV-infected macaques. In line with these observations, plasmacytoid dendritic cells have been shown to produce interferons upon SARS-CoV infection in vitro. Given the pivotal role of interferons during viral infections, (differential) induction of interferons may affect the outcome of the infection. Therefore, the functional implication of interferon production during SARS-CoV infection remains to be re-investigated.


title: Neutralizing antibodies to different proteins of African swine fever virus inhibit both virus attachment and internalization.

text: African swine fever virus induces in convalescent pigs antibodies that neutralized the virus before and after binding to susceptible cells, inhibiting both virus attachment and internalization. A further analysis of the neutralization mechanisms mediated by the different viral proteins showed that antibodies to proteins p72 and p54 are involved in the inhibition of a first step of the replication cycle related to virus attachment, while antibodies to protein p30 are implicated in the inhibition of virus internalization.


Paradoxical inflammation revisited: muraglitazar and cardiovascular risk.


title: The smallpox threat: the school nurse's role.

text: Today, with the threat of bioterrorism and war, there is a new dimension to the traditional role of the school nurse. The smallpox threat to public health will invoke the school nurse's role as an educator, liaison, and consultant in the community. This article discusses smallpox, the vaccination process, adverse effects, and postvaccination care. In addition to the role of educator, the school nurse has the role of a liaison between the school and the local health department. The school nurse also plays a vital role in school health policy development and implementation. Being prepared and educated makes our nation less vulnerable to these threats and will assist in keeping our children and communities safe.


title: Short polybasic peptide sequences are potent inhibitors of PC5/6 and PC7: Use of positional scanning-synthetic peptide combinatorial libraries as a tool for the optimization of inhibitory sequences.

text: Positional scanning-synthetic peptide combinatorial libraries (PS-SPCLs) are powerful molecular tools to identify enzyme substrate and potent inhibitory sequences and also to provide crucial information about active site determinants. PS-SPCLs have been surveyed for furin, proprotein convertase (PC)2, PC1/3, and PACE4 and proven efficient to identify potent peptidyl inhibitors in the low nanomolar range for furin and PC1/3. We report herein the screenings of nonamidated and acetylated hexapeptide PS-SPCLs for PC5/6A and PC7. The L-configuration library surveys distinctively revealed that L-Arg, L-Lys, and sometimes L-His in all six positions would generate the most potent inhibitors for both enzymes. Based on this clear polybasic preference, L-poly-Arg peptides ranging from four to nine residues were assayed. Inhibitory potency of these polybasic peptides increased with chain length, making nona-l-Arg a potent nanomolar inhibitor of PC5/6A and PC7 (Ki of 150 and 120 nM). PC5/6 and PC7 inhibition by nona-l-arginine was equivalent to that of furin (Ki of 114 nM) (J Biol Chem 275: 36741-36749, 2000). Nona-d-arginine was a more potent inhibitor of PC5/6 and PC7 than its levorotatory version (Ki of 19 and 81 nM), reminiscent of furin (Ki of 1.3 nM) (J Biol Chem 279:36788-36794, 2004). Our data indicate that certain poly-arginine peptides represent potent inhibitors targeting PCs of the constitutive secretory pathway (furin, PC5/6, and PC7). We conclude that basic residues within PC peptide inhibitors might be responsible for targeting PCs in general and for inhibitory potency, but that select amino acid changes will be necessary to acquire true specificity toward a single PC.


title: Capnography in the pediatric emergency department: clinical applications

text: Capnography, often referred to by emergency clinicians as end-tidal carbon dioxide monitoring, is a noninvasive method of measuring cardiopulmonary and metabolic parameters that can be utilized in many clinical applications. Growing evidence in the literature in support of the use of capnography has led to increased clinical use of this modality in many pediatric subspecialties. Understanding capnography and the literature supporting its practice will advance its use by emergency clinicians in the pediatric emergency department, promoting improved patient care and safety. This issue reviews the technology and physiology involved in measuring exhaled carbon dioxide and the interpretation of waveforms, and it highlights uses for capnography in pediatric patients in the emergency department. Uses include confirmation of intubation, maintenance of ventilation in intubated and nonintubated children, monitoring of effectiveness of cardiopulmonary resuscitation, and as an adjunct for monitoring of sedated children and children with lower respiratory disease and metabolic derangements.


title: Rapid onset of protection against infectious bovine rhinotracheitis with a modified-live virus multivalent vaccine.

text: This study provides evidence that subcutaneous vaccination of cattle with a commercially available modified-live virus combination vaccine can help reduce clinical signs associated with infectious bovine rhinotracheitis infections in feedlot animals vaccinated at the time of arrival. Calves vaccinated 72 or 96 hours before challenge had reduced clinical signs, lower body temperatures, lower virus titers, and 39% to 76% greater weight gains compared with nonvaccinated controls.


title: Initial outcomes of laparoscopic paraesophageal hiatal hernia repair with mesh.

text: The use of mesh in laparoscopic paraesophageal hiatal hernia repair (LHR) may reduce the risk of late hernia recurrence. The aim of this study was to evaluate initial outcomes and recurrence rate of 92 patients who underwent LHR reinforced with a synthetic bioabsorbable mesh. Surgical approaches included LHR and Nissen fundoplication (n = 64), LHR without fundoplication (n = 10), reoperative LHR (n = 9), LHR with a bariatric operation (n = 6), and emergent LHR (n = 3). The mean length of hospital stay was 2 ± 3 days (range, 1 to 30 days). There were no conversions to open laparotomy and no intraoperative complications. One of 92 patients (1.1%) required intensive care unit stay. The 90-day mortality was zero. Minor complications occurred in 3.3 per cent, major complications in 2.2 per cent, and late complications in 5.5 per cent of patients. There were no perforations or early hernia recurrence. The 30-day reoperation rate was 1.1 per cent. For patients with available 1-year follow-up, the overall recurrence rate was 18.5 per cent with a mean follow-up of 30 months (range, 12 to 51 months). LHR repair with mesh is associated with low perioperative morbidity and no mortality. The use of bioabsorbable mesh appears to be safe with no early hiatal hernia recurrence or late mesh erosion. Longer follow-up is needed to determine the long-term rate of hernia recurrence associated with LHR with mesh.


title: Efficacy of an inactivated vaccine against clinical disease caused by canine coronavirus.

text: Canine Coronavirus (CCV) is a causative agent of diarrhea in dogs. The reproduction of severe clinical disease with experimental CCV infection has been difficult. We have recently developed a CCV challenge model which reproduced clinical signs of disease in susceptible dogs. The following study was designed to determine whether immunization with an inactivated CCV vaccine would protect dogs from clinical disease induced using this model. Dogs (n = 13) were vaccinated with an inactivated CCV vaccine. Vaccinates and controls (n = 5) were orally inoculated with virulent CCV virus and treated with dexamethasone on days 0, 2, 4, and 6 after virus challenge. Control dogs developed clinical signs including diarrhea, dehydration, anorexia, depression, and nasal and ocular discharge. Diarrhea was noted in 80% of the controls and 60% progressed to a severe watery or bloody diarrhea that persisted for multiple days. Conversely, only 2/13 (15%) vaccinates developed mild diarrhea and none developed bloody diarrhea. The control dogs averaged 10.8 days of diarrhea compared to 1.4 days for vaccinates over the 21 day observation period. In addition to reduced clinical signs, the number of days of virus shedding and the level of CCV in feces was different for controls (100% shed virus) and vaccinates (38% shed virus). This study demonstrates that vaccination with an inactivated CCV vaccine can significantly reduce not only viral replication, but the occurrence of clinical disease following a virulent CCV infection.


title: System to assess genome sequencing needs for viral protein diagnostics and therapeutics.

text: Computational analyses of genome sequences may elucidate protein signatures unique to a target pathogen. We constructed a Protein Signature Pipeline to guide the selection of short peptide sequences to serve as targets for detection and therapeutics. In silico identification of good target peptides that are conserved among strains and unique compared to other species generates a list of peptides. These peptides may be developed in the laboratory as targets of antibody, peptide, and ligand binding for detection assays and therapeutics or as targets for vaccine development. In this paper, we assess how the amount of sequence data affects our ability to identify conserved, unique protein signature candidates. To determine the amount of sequence data required to select good protein signature candidates, we have built a computationally intensive system called the Sequencing Analysis Pipeline (SAP). The SAP performs thousands of Monte Carlo simulations, each calling the Protein Signature Pipeline, to assess how the amount of sequence data for a target organism affects the ability to predict peptide signature candidates. Viral species differ substantially in the number of genomes required to predict protein signature targets. Patterns do not appear based on genome structure. There are more protein than DNA signatures due to greater intraspecific conservation at the protein than at the nucleotide level. We conclude that it is necessary to use the SAP as a dynamic system to assess the need for continued sequencing for each species individually and to update predictions with each additional genome that is sequenced.


title: Severe acute respiratory syndrome (SARS): epidemiology and clinical features.

text: Severe acute respiratory syndrome (SARS) is a newly emerged infectious disease with a significant morbidity and mortality. The major clinical features include persistent fever, chills/rigor, myalgia, malaise, dry cough, headache, and dyspnoea. Older subjects may present without the typical febrile response. Common laboratory features include lymphopenia, thrombocytopenia, raised alanine transaminases, lactate dehydrogenase, and creatine kinase. The constellation of compatible clinical and laboratory findings, together with certain characteristic radiological features and lack of clinical response to broad spectrum antibiotics, should arouse suspicion of SARS. Measurement of serum RNA by real time reverse transcriptase-polymerase chain reaction technique has a detection rate of 75%-80% in the first week of the illness.


title: Frequency of enteropathogen detection in suckling and weaned pigs with diarrhea in Japan.

text: Fecal samples from suckling (n=153) and weaned (n=116) piglets with diarrhea in Japan were examined for shedding of viral, bacterial, and parasitic pathogens using culture, microscopic, and polymerase chain reaction methods. In suckling piglets, diarrhea was attributed to infection with a single etiologic agent in 60.8% of cases and with combinations of agents in 22.2%. In weaned piglets, diarrhea was attributed to a single etiologic agent in 43.1% and to combinations of agents in 47.4% of cases. Rotavirus was the most prevalent agent in suckling (67.3%) and weaned (65.5%) piglets. The detection of other pathogens was associated with age of the animals examined. Coccidia were predominantly isolated from suckling piglets, whereas Escherichia coli was found predominantly in weaned piglets. Although a relationship was not observed between detection rate of rotavirus and age of piglets, a single group of rotavirus was detected in 87.5% of suckling piglets whereas multiple groups were detected in 51.6% of weaned piglets. The results of this study confirm that diarrhea in piglets can, to a variable degree, be causally associated with multiple agents. Additionally, these results suggest reasons why this syndrome can be difficult to control.


COVID-19: An Unintended Force for Medical Revolution?


Humanized mice develop coronavirus respiratory disease.


Research agenda on persistent and unpredictable threat of influenza and emerging respiratory infections: a public health necessity in the Eastern Mediterranean Region.


title: Synthesis of cyclopentenyl carbocyclic nucleosides as potential antiviral agents against orthopoxviruses and SARS.

text: A practical and convenient methodology for the synthesis of chiral cyclopentenol derivative (+)-12a has been developed as the key intermediate that was utilized for the synthesis of biologically active carbocyclic nucleosides. The selective protection of allylic hydroxyl group followed by the ring-closing metathesis (RCM) reaction with Grubbs catalysts provided (+)-12a on a 10 g scale with 52% overall yield from D-ribose (4). The key intermediate (+)-12a was utilized for the synthesis of unnatural five-membered ring heterocyclic carbocyclic nucleosides. The newly synthesized 1,2,3-triazole analogue (17c) exhibited potent antiviral activity (EC(50) 0.4 microM) against vaccinia virus and moderate activities (EC(50) 39 microM) against cowpox virus and severe acute respiratory syndrome coronavirus (SARSCoV) (EC(50) 47 microM). The 1,2,4-triazole analogue (17a) also exhibited moderate antiviral activity (EC(50) 21 microM) against SARSCoV.


title: Laparoscopic repair of perineal hernia.

text: Perineal hernia is a rare but known complication following major pelvic surgery. It may occur spontaneously or following abdominoperineal resection, sacrectomy, or pelvic exenteration. Very little is known about spontaneous perineal hernia. Surgical repair via open transabdominal and transperineal approaches has been previously described. We report laparoscopic repair of spontaneous and postoperative perineal hernia in 2 patients.