title: Neural models for predicting viral vaccine targets.

text: We applied artificial neural networks (ANN) for the prediction of targets of immune responses that are useful for study of vaccine formulations against viral infections. Using a novel data representation, we developed a system termed MULTIPRED that can predict peptide binding to multiple related human leukocyte antigens (HLA). This implementation showed high accuracy in the prediction of the promiscuous peptides that bind to five HLA-A2 allelic variants. MULTIPRED is useful for the identification of peptides that bind multiple HLA-A2 variants as a group. By implementing ANN as a classification engine, we enabled both the prediction of peptides binding to multiple individual HLA-A2 molecules and the prediction of promiscuous binders using a single model. The ANN MULTIPRED predicts peptide binding to HLA-A*0205 with excellent accuracy (area under the receiver operating characteristic curve--AROC>0.90), and to HLA-A*0201, HLA-A*0204 and HLA-A*0206 with high accuracy (AROC>0.85). Antigenic regions with high density of binders ("antigenic hot-spots") represent best targets for vaccine design. MULTIPRED not only predicts individual 9-mer binders but also predicts antigenic hot spots. Two HLA-A2 hot-spots in Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) membrane protein were predicted by using MULTIPRED.


title: A practical guide for the diagnosis and treatment of pediatric pneumonia.

text: OBJECTIVE To develop guidelines for the diagnosis and management of community-acquired pediatric pneumonia. OPTIONS Clinical assessment, radiography, laboratory testing, and empirical antimicrobial therapy. OUTCOMES Increased awareness of age-related causes, improved accuracy of clinical diagnosis, better utilization of diagnostic testing and the rational use of empirical antimicrobial therapy resulting in more rapid diagnosis, initiation of appropriate therapy and decreased morbidity and mortality. EVIDENCE A MEDLINE search for relevant articles published from 1996 to September 1996 using the MeSH terms "pediatric," "pneumonia," "respiratory tract infection," "pneumonitis," "etiology," "diagnosis," "therapy," "antibiotics," "resistance," "radiology," "microbiology" and "biochemistry." VALUES A hierarchical evaluation of the strength of evidence modified from the methods of the Canadian Task Force on the Periodic Health Examination was used. When application of the hierarchy was not feasible or appropriate, different evaluation criteria were used. BENEFITS, HARMS AND COSTS Increased awareness of the causes of pneumonia, accurate diagnosis and prompt treatment should reduce costs associated with unnecessary investigations and complications due to inappropriate treatment. RECOMMENDATIONS Age is the best predictor of the cause of pediatric pneumonia, viral pneumonia being most common during the first 2 years of life. The absence of a symptom cluster of respiratory distress, tachypnea, crackles and decreased breath sounds accurately excludes the presence of pneumonia (level II evidence). Bacterial cultures of samples from the nasopharynx and throat have no predictive value; however, Gram staining and culture of sputum from older children and adolescents are useful (level III evidence). Oral antimicrobial therapy will provide adequate coverage for most mild to moderate forms of pneumonia in children (level III evidence). Parenteral therapy is typically reserved for neonates and patients with severe pneumonia admitted to hospital (level III evidence). VALIDATION These recommendations are based on consensus of Canadian experts in infectious diseases and microbiology. They are the only guidelines to address antimicrobial treatment from an age-related, etiologic perspective. SPONSOR The development of these guidelines and the technical support and assistance of Core Health Inc. in preparing this manuscript were funded through an unrestricted educational grant from Abbott Laboratories Canada. The sponsoring company was not involved in determining the membership of the consensus group or the content of the guidelines.


title: Pneumonia: classification, diagnosis and nursing management.

text: This article provides an overview of pneumonia as a high-incidence respiratory disease of varying severity in the 21st century. Many cases are mild to moderate and patients are successfully treated with antibiotics at home and with no lasting damage to the lungs. Vaccinations for influenza and, more recently, pneumococcal infections are becoming widely available for vulnerable groups of people, which will help to reduce the incidence of these diseases. However, pneumonia causes death in more severe cases with atypical forms such as Legionnaires' disease and severe acute respiratory syndrome (SARS) causing fatal outbreaks.


title: Hospital waste generation during an outbreak of severe acute respiratory syndrome in Taiwan.

text: During the SARS outbreak in Taiwan, the number of ambulatory patients and inpatients treated at one medical center decreased by 40%-70% because of the increasing number of SARS patients. At the peak of the epidemic, the amount of hospital infectious waste had increased from a norm of 0.85 kg per patient-day to 2.7 kg per patient-day. However, the hospital was able to return the generation of waste to normal levels within 10 days.


title: A rationale for using steroids in the treatment of severe cases of H5N1 avian influenza.

text: Acute hypercytokinaemia represents an imbalance of pro-inflammatory and anti-inflammatory cytokines, and is believed to be responsible for the development of acute respiratory distress syndrome and multiple organ failure in severe cases of avian (H5N1) influenza. Although neuraminidase inhibitors are effective in treating avian influenza, especially if given within 48 h of infection, it is harder to prevent the resultant hypercytokinaemia from developing if the patient does not seek timely medical assistance. Steroids have been used for many decades in a wide variety of inflammatory conditions in which hypercytokinaemia plays a role, such as sepsis and viral infections, including severe acquired respiratory syndromes and avian influenza. However, to date, the results have been mixed. Part of the reason for the discrepancies might be the lack of understanding that low doses are required to prevent mortality in cases of adrenal insufficiency. Adrenal insufficiency, as defined in the sepsis/shock literature, is a plasma cortisol rise of at least 9 microg dl(-1) following a 250 microg dose of adrenocorticotropin hormone (ACTH), or reaching a plasma cortisol concentration of >25 microg dl(-1) following a 1-2 microg dose of ACTH. In addition, in the case of hypercytokinaemia induced by potent viruses, such as H5N1, systemic inflammation-induced, acquired glucocorticoid resistance is likely to be present. Adrenal insufficiency can be overcome, however, with prolonged (7-10 or more days) supraphysiological steroid treatment at a sufficiently high dose to address the excess activation of NF-kappaB, but low enough to avoid immune suppression. This is a much lower dose than has been typically used to treat avian influenza patients. Although steroids cannot be used as a monotherapy in the treatment of avian influenza, there might be a potential role for their use as an adjunct treatment to antiviral therapy if appropriate dosages can be determined. In this paper, likely mechanisms of adrenal insufficiency are discussed, drawing from a broad background of literature sources.


Laparoendoscopic single site splenectomy.


Doxepin increases serum cholesterol levels.


Going with the Flow: Covid-19 Pathway and Experience in Galway University Hospital.


title: Tumor necrosis factor-alpha convertase (ADAM17) mediates regulated ectodomain shedding of the severe-acute respiratory syndrome-coronavirus (SARS-CoV) receptor, angiotensin-converting enzyme-2 (ACE2).

text: Angiotensin-converting enzyme-2 (ACE2) is a critical regulator of heart function and a cellular receptor for the causative agent of severe-acute respiratory syndrome (SARS), SARS-CoV (coronavirus). ACE2 is a type I transmembrane protein, with an extracellular N-terminal domain containing the active site and a short intracellular C-terminal tail. A soluble form of ACE2, lacking its cytosolic and transmembrane domains, has been shown to block binding of the SARS-CoV spike protein to its receptor. In this study, we examined the ability of ACE2 to undergo proteolytic shedding and investigated the mechanisms responsible for this shedding event. We demonstrated that ACE2, heterologously expressed in HEK293 cells and endogenously expressed in Huh7 cells, undergoes metalloproteinase-mediated, phorbol ester-inducible ectodomain shedding. By using inhibitors with differing potency toward different members of the ADAM (a disintegrin and metalloproteinase) family of proteases, we identified ADAM17 as a candidate mediator of stimulated ACE2 shedding. Furthermore, ablation of ADAM17 expression using specific small interfering RNA duplexes reduced regulated ACE2 shedding, whereas overexpression of ADAM17 significantly increased shedding. Taken together, these data provided direct evidence for the involvement of ADAM17 in the regulated ectodomain shedding of ACE2. The identification of ADAM17 as the protease responsible for ACE2 shedding may provide new insight into the physiological roles of ACE2.


title: A Case of Pediatric Multisystem Inflammatory Syndrome Temporally Associated with COVID-19 in South Dakota.

text: Coronavirus infectious disease (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was deemed a worldwide pandemic by the World Health Organization in February 2020. The U.S. began seeing epidemic levels of cases in early March 2020. South Dakota case numbers dramatically increased in late March/early April, 2020 due to a large meat processing facility outbreak. Although COVID-19 infections in adults more severely involve the lungs and heart with multiple organ-system dysfunction, pediatric patients have largely been spared. In May 2020, a syndrome resembling severe Kawasaki disease with shock in children was reported from European groups. We report a case that presented to and was managed in our Sioux Falls pediatric intensive care unit in April 2020 that fits the description, course, and successful treatment described by our European colleagues. Our case fulfils the case definition of paediatric multisystem inflammatory syndrome temporally associated with COVID-19 described by the Royal College of Paediatrics and Child Health on April 27, 2020. We will review and discuss the European and US case definitions of this syndrome and similarities, and differences with Kawasaki disease and treatment options.


title: COVID-19 and Management of Orthopedic Emergencies. Producing a Consensus of Experts.

text: OBJECTIVE Report the results of a participatory approach in Tunisian orthopedic surgery, for the development of a consensus of experts, on the identification of the list of pathologies to always be considered as non-postponable emergencies, during the COVID-19 pandemic. MATERIAL AND METHODS This descriptive study of the opinions of Tunisian experts covered all orthopedic morbidities classified into three homogeneous groups: trauma, infections and tumors of the musculoskeletal system. The attitudes of the interviewees were collected using the "Delphi" method, using a "Google-Form" questionnaire, sent by email to all of the Tunisian university orthopedic surgeons in practice, and registered at the SOTCOT. Consensus has been established for an item, if validated by at least 80% of the experts. The analysis of the results focused on the first 30 responses to this "online" form. RESULTS Tunisian experts agreed on the continuity of the urgency of taking in charge all the items of orthopedic morbidity during the COVID-19 pandemic, apart from the following affections: aseptic nonunions in the upper and lower limbs, aggressive giant cell tumors, and hyperalgesic disc herniations, where agreement rates were only at 8%, 12%, 58% and 77%. CONCLUSION Relative to its disciplinary and professional specificities, the majority the of orthopedic conditions were still considered as emergencies, during the COVID-19 pandemic, which did not lend to postponement of the surgery. However, their management should obey to the recommendations of "Sorting" and the " COVID-19 Patient Pathway ", established by national authorities.


title: Human bocavirus in Italian patients with respiratory diseases.

text: BACKGROUND hBoV, a recently discovered parvovirus, can be present in the respiratory tract of patients with acute respiratory diseases (ARD), but its etiologic involvement in the underlying diseases is still uncertain. OBJECTIVE To determine in a retrospective study, the prevalence of hBoV, compared with common respiratory viruses (RV), in respiratory specimens from patients with ARD. STUDY DESIGN A total of 335 specimens obtained over 7 years were examined. Two hundred were nasal swabs from infants hospitalized for ARD, 84 were nasal swabs or bronchoalveolar lavages from adults with pneumonia, bronchopneumonia or asthma, and 51 were nasal swabs from healthy children. RESULTS The overall rate of hBoV detection in specimens from infants with ARD, which was 4.5%, varied slightly from year to year, except for the period 2000-2002, when no specimen was positive. Unlike other RV, no seasonal variation in hBoV incidence was noted. Infants with hBoV infection suffered either from bronchiolitis or from bronchopneumonia and 5 out of 9 cases yielded no co-infecting viral pathogen. Only one sample from an adult was hBoV positive. None of the nasal swabs from healthy subjects tested hBoV-positive. CONCLUSIONS The findings indicate that hBoV can cause ARD in infants.


title: [Recommendations for the use of thromboprophylaxis in hospitalized patients with COVID-19 in Argentina].

text: Although the incidence is uncertain, some case reports suggest that COVID 19 infection is associated with an increased risk of venous thromboembolism. We suggest starting prophylactic anticoagulant therapy for all patients hospitalized with a symptomatic infection with COVID-19, unless contraindicated, with enoxaparin 40 mg SC daily if creatinine clearance is greater than 30 ml/min.


title: Global surveillance for chemical incidents of international public health concern.

text: OBJECTIVE In December 2001, an expert consultation convened by WHO identified strengthening national and global chemical incident preparedness and response as a priority. WHO is working towards this objective by developing a surveillance and response system for chemical incidents. This report describes the frequency, nature and geographical location of acute chemical incidents of potential international concern from August 2002 to December 2003. METHODS Acute chemical incidents were actively identified through several informal (e.g. Internet-based resources) and formal (e.g. various networks of organizations) sources and assessed against criteria for public health emergencies of international concern using the then proposed revised International Health Regulations (IHR). WHO regional and country offices were contacted to obtain additional information regarding identified incidents. FINDINGS Altogether, 35 chemical incidents from 26 countries met one or more of the IHR criteria. The WHO European Region accounted for 43% (15/35) of reports. The WHO Regions for Africa, Eastern Mediterranean and Western Pacific each accounted for 14% (5/35); South-East Asia and the Americas accounted for 9% (3/35) and 6% (2/35), respectively. Twenty-three (66%) events were identified within 24 hours of their occurrence. CONCLUSION To our knowledge this is the first global surveillance system for chemical incidents of potential international concern. Limitations such as geographical and language bias associated with the current system are being addressed. Nevertheless, the system has shown that it can provide early detection of important events, as well as information on the magnitude and geographical distribution of such incidents. It can therefore contribute to improving global public health preparedness.


Covid-19: The Pandemic of Our Time


title: Self-reported visual impairment among persons with diagnosed diabetes --- United States, 1997--2010.

text: Diabetes can lead to visual impairment (VI) and blindness (1). However, early detection and treatment of many common eye diseases, such as diabetic retinopathy and glaucoma, can reduce the risk for developing VI (1). Surveillance of VI among persons with diabetes is important for evaluating the effectiveness of efforts to reduce VI and other complications of diabetes. To examine trends in the prevalence of self-reported VI among adults (persons aged ≥18 years) with diagnosed diabetes in the United States and to assess reported access to eye-care providers, CDC analyzed 1997-2010 data from the National Health Interview Survey (NHIS). This report describes the results of that analysis, which indicated that although the number of adults with diagnosed diabetes reporting VI increased, the age-adjusted percentage of adults with diagnosed diabetes who reported VI declined significantly, from 23.7% in 1997 to 16.7% in 2010. During this 14-year period, age-adjusted VI prevalence declined significantly among most categories of adults with diabetes: men, women, whites, Hispanics, those with some college or higher education, and those diagnosed with diabetes for ≥3 years. Prevalence also declined among those aged ≥45 years. The percentage of adults with diagnosed diabetes and self-reported VI who reported having consulted an eye-care provider in the past year remained constant at approximately 63%. Continued efforts are needed to sustain and improve the declining trends in self-reported VI and to increase rates of recommended eye examinations in the population with diabetes.


title: Protecting public health in the age of emerging infections.

text: Emerging and re-emerging infections cause huge concern among public health workers and international and national bodies such as the World Health Organization (WHO) and the U.S. National Institutes of Health (NIH). Indeed, scientists around the world express the view that despite the danger, research on these emerging virulent pathogens is crucial and must continue. While most of the studies underway are targeted at improving and protecting public health, some studies bear potentiallyserious risks resulting from misuse. These studies are defined as dual-use research of concern (DURC), where it is not evident that the benefits outweigh the risks. The H5N1 controversy has pushed various governments to institute new policies to govern such research. We describe the regulations that govern this emerging field of research in the United States and Israel, two countries that have taken leading stands on these issues. We suggest that the existing policies are able to mitigate many of the risks that this research encapsulates, yet more work is required--especially on the global level.


Imaging of Covid-19; an Irish Perspective


title: Assembling the SARS-CoV genome -- new method based on graph theoretical approach.

text: Nowadays, scientists may learn a lot about the organisms studied just by analyzing their genetic material. This requires the development of methods of reading genomes with high accuracy. It has become clear that the knowledge of the changes occurring within a viral genome is indispensable for effective fighting of the pathogen. A good example is SARS-CoV, which was a cause of death of many people and frightened the entire world with its fast and hard to prevent propagation. Rapid development of sequencing methods, like shotgun sequencing or sequencing by hybridization (SBH), gives scientists a good tool for reading genomes. However, since sequencing methods can read fragments of up to 1000 bp only, methods for sequence assembling are required in order to read whole genomes. In this paper a new assembling method, based on graph theoretical approach, is presented. The method was tested on SARS-CoV and the results were compared to the outcome of other widely known methods.


title: Specific mutations in a viral RNA pseudoknot drastically change ribosomal frameshifting efficiency.

text: Many viruses regulate protein synthesis by -1 ribosomal frameshifting using an RNA pseudoknot. Frameshifting is vital for viral reproduction. Using the information gained from the recent high-resolution crystal structure of the beet western yellow virus pseudoknot, a systematic mutational analysis has been carried out in vitro and in vivo. We find that specific nucleotide tertiary interactions at the junction between the two stems of the pseudoknot are crucial. A triplex is found between stem 1 and loop 2, and triplex interactions are required for frameshifting function. For some mutations, loss of one hydrogen bond is sufficient to abolish frameshifting. Furthermore, mutations near the 5' end of the pseudoknot can increase frameshifting by nearly 300%, possibly by modifying ribosomal contacts. It is likely that the selection of suitable mutations can thus allow viruses to adjust frameshifting efficiencies and thereby regulate protein synthesis in response to environmental change.


title: West Nile virus-induced cytoplasmic membrane structures provide partial protection against the interferon-induced antiviral MxA protein.

text: The human MxA protein is a type I and III interferon (IFN)-induced protein with proven antiviral activity against RNA viruses. In this study, we investigated the effect of MxA expression on the replication of West Nile Virus strain Kunjin (WNV(KUN)). Pretreatment of A549 cells with IFN-alpha lead to increased expression of MxA, which contributed to inhibition of WNV(KUN) replication and secretion. However, in Vero cells stably expressing the MxA protein, WNV(KUN) replication, maturation and secretion was not inhibited. Biochemical and subcellular localization studies of WNV(KUN) proteins and MxA suggest that the MxA activity was not compromised by a flavivirus-encoded antagonist. Instead, we show that characteristic membranous structures induced during WNV(KUN) replication provide partial protection from MxA, possibly by 'hiding' WNV(KUN) replication components. This distinct compartmentalization of viral replication and components of the cellular antiviral response may be an evolutionary mechanism by which flaviviruses can hide from host surveillance.


title: [Retinoic acid modification of cell culture used for reproduction of enteropathogenic viruses].

text: The 0.001-0.005% retinoic acid injection into the growth cultural medium of prime and continue cell cultures 12-24h before inoculation considerably raised the cell sensitivity to animal entero- and coronaviruses. The entero- and coronaviruses concentrations in cultural medium increased by 10(1.58) and 10(1.68)TCID 50/1.0 respectively. The optimized parameters of the cell culture processing for the enteropathogenic viruses reproduction improvement are proposed.


title: Antigenic relationship of turkey coronavirus isolates from different geographic locations in the United States.

text: The purpose of the present study was to examine the antigenicity of turkey coronavirus (TCV) isolates from various geographic areas with antibodies to different viruses. Seventeen isolates of TCV were recovered from intestinal samples submitted to Animal Disease Diagnostic Laboratory, Purdue University, from turkey farms located in different geographic areas. The prototype TCV Minnesota isolate (TCV-ATCC) was obtained from the American Type Culture Collection. Intestinal sections were prepared from turkey embryos infected with different TCV isolates and reacted with polyclonal or monoclonal antibodies to TCV, infectious bronchitis virus (IBV), bovine coronavirus (BCV), transmissible gastroenteritis virus (TGEV), reovirus, rotavirus, adenovirus, or enterovirus in immunofluorescent antibody staining. All 18 TCV isolates have the same antigenic reactivity pattern with the same panel of antibodies. Positive reactivity was seen with polyclonal antibodies to the TCV Indiana isolate, the TCV Virginia isolate, TCV-ATCC, and the IBV Massachusetts strain as well as monoclonal antibodies to the TCV North Carolina isolate or the membrane protein of IBV. Antibodies to BCV or TGEV were not reactive with any of the TCV isolates. Reactivity of antibodies to unrelated virus, rotavirus, reovirus, adenovirus, or enterovirus with different TCV isolates was all negative, except positive response was seen between enterovirus antibody and a TCV western North Carolina isolate, suggesting coinfection of turkeys with TCV and enterovirus in that particular case. The results indicated that the TCV isolates from these geographic locations in the U.S. shared close antigenicity and were antigenically related to IBV.


title: Assessment the Efficiency of the Constructed Minigenome of Rabies Virus using PV Strain as Helper Virus.

text: INTRODUCTION Rabies is an acute viral disease that causes encephalomyelitis in mammals and human. The only way to prevent this disease is through vaccination before or after exposure. The aim of this study is to evaluate the efficiency of the Pasteur virus (PV) minigenome, using PV strain. MATERIALS AND METHODS Enhanced Green Fluorescent Protein (EGFP) sequence was placed between the designed necessary elements (Hammerhead, HDV ribozyme, 3' Leader, and 5' Trailer sequences), which resemble the rabies virus PV strain (PV2061) genome and anti-genome. These constructs were placed between T7 polymerase promoter and T7 polymerase terminator sequences. The accuracy of the minigenome was confirmed by the expression of EGFP using the helper virus in T7-BHK cell line. RESULTS The viral necessary elements of positive and negative sense strands were evaluated for the ability of EGFP expression in the presence of the helper virus. While the positive strand showed background results, no EGFP background was observed in the negative strand application. CONCLUSION Establishment of minigenome system does not require advanced biosafety levels. Furthermore, using minigenome system eliminates many potential confounding factors that may be present in coding regions of the genome. Use of the minigenome system is easier and more feasible than the full genome rescue of the virus. This study successfully shows the efficiency of the constructed rabies virus minigenome in expression of inserted gene.


Utilization of the TAMIS technique for trans-stomal excision of a colonic neoplasm: a video demonstration.


title: Onyx Embolization of a Meningioma with a Dysplastic Aneurysmal Anterior Cerebral Artery Vessel.

text: Preoperative embolization of meningiomas can be safely performed using a variety of embolic agents. Most commonly, the vascular supply from branches of the external carotid artery is the target of embolization. In our report, we detail the treatment of a patient with a parafalcine meningioma that received its supply via branches of the anterior cerebral artery. One of the feeder vessels appeared to contain a dysplastic aneurysmal dilatation of the vessel. Due to patient circumstances, embolization was performed using standard microcatheterization techniques to minimize intraoperative blood loss. We report a rare instance of endovascular treatment of a pial vessel to treat an intracranial meningioma using Onyx.


title: High-frequency leader sequence switching during coronavirus defective interfering RNA replication.

text: A system was developed that exploited defective interfering (DI) RNAs of coronavirus to study the role of free leader RNA in RNA replication. A cDNA copy of mouse hepatitis virus DI RNA was placed downstream of the T7 RNA polymerase promoter to generate DI RNAs capable of extremely efficient replication in the presence of a helper virus. We demonstrated that, in the DI RNA-transfected cells, the leader sequence of these DI RNAs was switched to that of the helper virus during one round of replication. This high-frequency leader sequence exchange was not observed if a nine-nucleotide stretch of sequence (UUUAUAAAC) at the junction between the leader and the remaining DI sequence was deleted. This observation suggests that a free leader RNA generated from the genomic RNA of mouse hepatitis virus may participate in the replication of DI RNA.


title: Structural and functional basis for ADP-ribose and poly(ADP-ribose) binding by viral macro domains.

text: Macro domains constitute a protein module family found associated with specific histones and proteins involved in chromatin metabolism. In addition, a small number of animal RNA viruses, such as corona- and toroviruses, alphaviruses, and hepatitis E virus, encode macro domains for which, however, structural and functional information is extremely limited. Here, we characterized the macro domains from hepatitis E virus, Semliki Forest virus, and severe acute respiratory syndrome coronavirus (SARS-CoV). The crystal structure of the SARS-CoV macro domain was determined at 1.8-Angstroms resolution in complex with ADP-ribose. Information derived from structural, mutational, and sequence analyses suggests a close phylogenetic and, most probably, functional relationship between viral and cellular macro domain homologs. The data revealed that viral macro domains have relatively poor ADP-ribose 1"-phosphohydrolase activities (which were previously proposed to be their biologically relevant function) but bind efficiently free and poly(ADP-ribose) polymerase 1-bound poly(ADP-ribose) in vitro. Collectively, these results suggest to further evaluate the role of viral macro domains in host response to viral infection.


title: Severe acute respiratory syndrome and its effects on health care: how Canada has dealt with this ordeal.

text: The global epidemic of severe acute respiratory syndrome (SARS) indirectly has affected all health care facilities in affected nations and has unaffected nations on alert. Health care practitioners need to be aware of infection control measures to prevent the spread of SARS. This article looks at this new disease and how it has affected the delivery of health care in Canada.


title: Intensive care ventilators.

text: Intensive care ventilators provide temporary ventilatory support to critically ill patients who cannot breathe on their own or who require assistance to maintain adequate ventilation. These ventilators offer a wide range of capabilities and can meet the needs of many types of patients. We last evaluated intensive care ventilators for adult use in our July-August 2000 issue. In the current Evaluation, we revisit the ratings of nine previously evaluated ventilators and describe our findings for one newly evaluated unit, the Siemens Servo-i. As ventilator technology has evolved, so have our ratings. Several units have added new features and capabilities since we last examined them. As a result, we have rated them higher in this study. Conversely, a few models have been given lower ratings because they haven't kept up with the advances in the technology; although they perform adequately, we can no longer recommend their purchase because better choices are available.


title: [Adaptation of an isolate of bovine enteritic coronavirus to serial growth in cell culture and characterization of the virus (author's transl)].

text: Coronaviruses are known to behave as pathogens in a variety of animal species. Their multiplication in the differentiated enterocytes of the intestinal tract induces a severe diarrheic syndrome which may lead to death. To study these viruses and to progress in the development of prophylactic and diagnostic methods, it is essential to succeed in adapting the agents to serial growth in cell culture. In spite of several attempts adaptation to primary cell cultures could be obtained in the past only for two isolates. The present work describes the methodology which enabled us to succeed in the adaptation of a bovine enteric coronavirus isolated in France to primary calf kidney cells, and reports results on the characteristics of this virus. Adaptation to serial growth was realized at 34 degrees C in the following way: 3 passages each lasting 8 to 10 days followed by 16 passages each lasting 6 days. From the 20th passage on the duration of incubation was reduced to 4 days. Virus growth was checked using an indirect immunofluorescence test. During the first passages only a few dispersed positive cells could be observed but later on small foci of immunofluorescent cells did appear. By the 35th passage infective titers did not exceed 1 x 10(5) TCID50/ml and cytopathic effects were always very discrete. When the susceptibility of different cell-types to this coronavirus strain (G110) was checked we found that only bovine cells and the human HRT18 cell-line were able to replicate the virus. In HRT18 cells titers as high as 5 x 10(7) TCID50/ml could be obtained at the first passage of G110 strain, previously adapted to primary calf kidney cells. As regards the antigenic properties of G110 virus no difference could be established between this strain and the Nebraska isolate (NCDCV) neither by indirect immunofluorescence test nor by indirect neutralization test. The G110 strain agglutinates rat and mouse erythrocytes. The density of purified virions in mouse gradients is 1.19 and the morphology of the particles is characteristic of a coronavirus.


title: A conserved coronavirus epitope, critical in virus neutralization, mimicked by internal-image monoclonal anti-idiotypic antibodies.

text: Monoclonal antibody (MAb) 6A.C3 neutralizes transmissible gastroenteritis coronavirus (TGEV) and is specific for a conserved epitope within subsite Ac of the spike (S) glycoprotein of TGEV. Six hybridomas secreting anti-idiotypic (Ab2) MAbs specific for MAb 6A.C3 (Ab1) have been selected. All six MAbs inhibited the binding of Ab1 to TGEV and specifically cross-linked MAb1-6A.C3. Four of these hybridomas secreted gamma-type anti-idiotypic MAbs. The other two Ab2s (MAbs 9A.G3 and 9C.E11) were recognized by TGEV-specific antiserum induced in two species. This binding was inhibited by viruses of the TGEV group but not by serologically unrelated coronaviruses. These results indicate that MAb2-9A.G3 and MAb2-9C.E11 mimic an antigenic determinant present on the TGEV surface, and they were classified as beta-type ("internal-image") MAbs. TGEV-binding Ab3 antiserum was induced in 100% of mice immunized with the two beta-type MAb2s and in 25 to 50% of mice immunized with gamma-type MAb2. Both beta- and gamma-type Ab2s induced neutralizing Ab3 antibodies in mice that were mainly directed to antigenic subsite Ac of the S protein.


title: The impact of the SARS outbreak on an urban emergency department in Taiwan.

text: OBJECTIVE Emergency departments (ED) were on the front lines for possible cases of transmission during the severe acute respiratory syndrome (SARS) epidemic. The purpose of this study was to investigate the impact of the SARS catastrophe on an urban ED. METHODS The patients' characteristics in an urban ED were collected from March to May 2003 during the SARS outbreak in Taiwan. The crisis period was divided into 2 periods: 30 days before (period 1) and after (period 2) April 21, the date of the first hospital-associated transmission. Problem severity in the ED and stress levels of ED staff during the SARS catastrophe were rated from mild (1 point) to severe (5 points). RESULTS The number of ED patients declined 33.4% in period 2. There was a 2.1% (95%CI, 0.4-3.8) increase in the percentage of male patients, a 2.5% (95% CI, 1.5-3.7) increase in percentage of fever (>38 degrees C), and a 4.0% (95% CI, 2.6-5.4%) increase in chief complaint of fever in period 2. The number of nontrauma patients younger than 18 years had declined by 44.5% in period 2. The total charge for reimbursement from an insurance institution declined 21.7%. During the SARS outbreak, the most severe stress experienced by either physicians or nurses occurred during emergency resuscitation (median stress rating point, 4; interquartile range, 1). CONCLUSION The SARS catastrophe affected the ED visit volume, finances, various patient characteristics, and stress levels in the ED physicians and nurses. EDs must be fully prepared to face the challenges of the next outbreak of SARS or other infectious disease.


Performance of transport ventilators.


Analysis of CNS inflammatory responses to MHV. Role of spike determinants in initiating chemokine and cytokine responses.


title: Characterization and isolation of structural polypeptides in haemagglutinating encephalomyelitis virus.

text: Haemagglutinating encephalomyelitis virus (HEV), a member of the coronavirus family, was purified and analysed by SDS-polyacrylamide gel electrophoresis. It was shown to contain eight polypeptides, seven of which were glycosylated. They had apparent mol. wt. of 180,000 (GP 180), 130,0000 (GP 130), 120,000 (GP 120) 76,000 (GP 76), 64,000 (VP 64), 54,000 (GP 54), 32,000 (GP 32) and 31,000 (GP 31). Electrophoresis of virus samples dissociated under varying conditions showed that GP 54 and GP 120 could be interpreted as larger products of GP 31 and GP 32 and of GP 76, respectively. GP 76 also appeared as a dimer with a mol. wt. of 140,000 (GP 140) in the absence of beta-mercaptoethanol. Subviral particles, obtained by treatment with bromelain, banded at a slightly lower density than the intact virus and lacked surface projections. Analysis of these particles indicated that GP 180, GP 130 and GP 76 are associated with the virus projections. A small part of GP 31 and GP 32 also appeared to protrude from the lipid envelope since 20% of each molecule was sensitive to digestion. Two glycoproteins, GP 130 and GP 76, were solubilized with the detergent Triton X-100 and separated by rate zonal centrifugation. According to its activity-in indirect haemagglutination tests, GP 76 was considered to be a monovalent haemagglutinin subunit.


title: Antibodies to some swine diseases in commercial piggeries in Central Zambia.

text: Blood samples were taken from 121 sows and gilts on 7 commercial piggeries located around Lusaka (Zambia). The samples tested negative for antibodies to Aujeszky's disease, transmissible gastroenteritis (TGE), swine influenza, hog cholera and brucellosis. Seventy-eight pigs from 5 farms had positive titres to porcine parvovirus. Eighteen sera showed positive titres to Leptospira celledoni.


title: Detection of a novel and highly divergent coronavirus from asian leopard cats and Chinese ferret badgers in Southern China.

text: Since an outbreak of severe acute respiratory syndrome (SARS) was averted in 2004, many novel coronaviruses have been recognized from different species, including humans. Bats have provided the most diverse assemblages of coronaviruses, suggesting that they may be the natural reservoir. Continued virological surveillance has proven to be the best way to avert this infectious disease at the source. Here we provide the first description of a previously unidentified coronavirus lineage detected from wild Asian leopard cats (Prionailurus bengalensis) and Chinese ferret badgers (Melogale moschata) during virological surveillance in southern China. Partial genome analysis revealed a typical coronavirus genome but with a unique putative accessory gene organization. Phylogenetic analyses revealed that the envelope, membrane, and nucleoprotein structural proteins and the two conserved replicase domains, putative RNA-dependent RNA polymerase and RNA helicase, of these novel coronaviruses were most closely related to those of group 3 coronaviruses identified from birds, while the spike protein gene was most closely related to that of group 1 coronaviruses from mammals. However, these viruses always fell into an outgroup phylogenetic relationship with respect to other coronaviruses and had low amino acid similarity to all known coronavirus groups, indicating that they diverged early in the evolutionary history of coronaviruses. These results suggest that these viruses may represent a previously unrecognized evolutionary pathway, or possibly an unidentified coronavirus group. This study demonstrates the importance of systematic virological surveillance in market animals for understanding the evolution and emergence of viruses with infectious potential.


Micro total analysis systems. Latest advancements and trends.


Anxiety and Morale in Front-Line Healthcare Workers During the Coronavirus Disease 2019 (COVID-19) Outbreak at the National Screening Centre in Singapore.


title: Epidemic of diarrhoea caused by porcine epidemic diarrhoea virus in Italy.

text: There was an epidemic of diarrhoea affecting pigs of all ages in Italy between May 2005 and June 2006. In 63 herds the cause was confirmed as porcine epidemic diarrhoea virus by electron microscopy, immunoelectron microscopy, pcr and serology. Watery diarrhoea without mucus and blood was usually associated with a reduction of feed consumption. In farrowing-to-weaning herds, diarrhoea affected the sows and suckling piglets, and the mortality in newborn piglets was up to 34 per cent. In growers and fatteners the morbidity ranged from 20 to 80 per cent, but there was either no mortality or it was very low. Depending on the size of the herd and the type of operation, the clinical disease lasted for weeks or months.


title: Characterization of cytokine/chemokine profiles of severe acute respiratory syndrome.

text: RATIONALE There is currently no optimal treatment or effective drug for severe acute respiratory syndrome (SARS), because the immunopathologic mechanism is poorly understood. OBJECTIVES To explore the immune mechanism underlying the pathogenesis of SARS, we studied the expression profile of cytokines/chemokines in the blood and the immunopathology of the lung and lymphoid tissues. METHODS Fourteen cytokines/chemokines in the blood of 23 patients with SARS were dynamically screened, using a bead-based multiassay system. Reverse transcription-polymerase chain reaction was performed to amplify mRNA. Histopathology of the lung and lymphoid tissues at autopsy was examined, using methods of immunohistochemistry and double immunofluorescence staining. MAIN RESULTS Interferon-inducible protein-10 (IP-10) was markedly elevated in the blood during the early stage of SARS, and remained at a high level until convalescence. Moreover, IP-10 was highly expressed in both lung and lymphoid tissues, where monocyte-macrophage infiltration and depletion of lymphocytes were observed. The levels of interleukin-6, interleukin-8, and monocyte chemoattractant protein-1 were concomitantly increased in the blood of the patients with superinfection, and the mRNAs for these cytokines were also increased in lung tissues. CONCLUSIONS Induction of IP-10 is a critical event in the initiation of immune-mediated acute lung injury and lymphocyte apoptosis during the development of SARS. Superinfection after the immune injury is the main cause of death. The prompt elevation of interleukin-6, interleukin-8, and monocyte chemoattractant protein-1 is a sign of superinfection, indicating a high risk of death.


Comments from the Center for Biosecurity of UPMC on the National Strategy for Pandemic Influenza: Implementation Plan.


title: Why own an exotic pet?

text: Even though people have owned a wide variety of companion animals since times of old, the modern craze for increasingly exotic and little-known species raises a number of questions, including some of an ethical nature. While trade in exotic animals is certainly profitable for these who practise it, it poses great risks of varying types: ecological risks, threats to biodiversity conservation and health risks. Several introduced animal populations have gone on to establish a line in their new host country. We are just starting to measure the adverse impact this has had, in some cases on a very large scale. The veterinary profession doubtless has a major role to play in endeavouring to reform this trade in living creatures that unfortunately results in many losses.


title: Effect of membrane curvature-modifying lipids on membrane fusion by tick-borne encephalitis virus.

text: Enveloped viruses enter cells by fusion of their own membrane with a cellular membrane. Incorporation of inverted-cone-shaped lipids such as lysophosphatidylcholine (LPC) into the outer leaflet of target membranes has been shown previously to impair fusion mediated by class I viral fusion proteins, e.g., the influenza virus hemagglutinin. It has been suggested that these results provide evidence for the stalk-pore model of fusion, which involves a hemifusion intermediate (stalk) with highly bent outer membrane leaflets. Here, we investigated the effect of inverted-cone-shaped LPCs and the cone-shaped oleic acid (OA) on the membrane fusion activity of a virus with a class II fusion protein, the flavivirus tick-borne encephalitis virus (TBEV). This study included an analysis of lipid mixing, as well as of the steps preceding or accompanying fusion, i.e., binding to the target membrane and lipid-induced conformational changes in the fusion protein E. We show that the presence of LPC in the outer leaflet of target liposomes strongly inhibited TBEV-mediated fusion, whereas OA caused a very slight enhancement, consistent with a fusion mechanism involving a lipid stalk. However, LPC also impaired the low-pH-induced binding of a soluble form of the E protein to liposomes and its conversion into a trimeric postfusion structure that requires membrane binding at low pH. Because inhibition is already observed before the lipid-mixing step, it cannot be determined whether impairment of stalk formation is a contributing factor in the inhibition of fusion by LPC. These data emphasize, however, the importance of the composition of the target membrane in its interactions with the fusion peptide that are crucial for the initiation of fusion.


title: Preparation of His-tagged armored RNA phage particles as a control for real-time reverse transcription-PCR detection of severe acute respiratory syndrome coronavirus.

text: Armored RNA has been increasingly used as both an external and internal positive control in nucleic acid-based assays for RNA virus. In order to facilitate armored RNA purification, a His6 tag was introduced into the loop region of the MS2 coat protein, which allows the exposure of multiple His tags on the surface during armored RNA assembly. The His-tagged armored RNA particles were purified to homogeneity and verified to be free of DNA contamination in a single run of affinity chromatography. A fragment of severe acute respiratory syndrome coronavirus (SARS-CoV) genome targeted for SARS-CoV detection was chosen for an external positive control preparation. A plant-specific gene sequence was chosen for a universal noncompetitive internal positive control preparation. Both controls were purified by Co2+ affinity chromatography and were included in a real-time reverse transcription-PCR assay for SARS-CoV. The noncompetitive internal positive control can be added to clinical samples before RNA extraction and enables the identification of potential inhibitive effects without interfering with target amplification. The external control could be used for the quantification of viral loads in clinical samples.


Assessment of new enzyme immunoassay to detect herpes simplex virus antigen.


title: Transanal endoscopic microsurgery for rectal tumours using a Single Incision Laparoscopic Port.

text: BACKGROUND Transanal endoscopic microsurgery (TEM), first described by Buess enables a less aggressive approach of benign rectal lesions, or even early rectal cancer in a curative intent. MATERIALS AND METHODS The SILS Port, initially designed for laparoscopic surgery, was successfully used for 20 TEM procedures in 16 patients. Local resection was sufficient in 15 procedures (benign tumours or pT1) out of 20, whereas 5 TEM operations required additional surgery: 3 rectal resections (pT1Nx, pT1sm3Nx and pT2N1) and 1 TEM revision (1 patient refused the rectal resection (pT1Nx). Postoperative complications following Dindo-Clavien were: grade II in 4 patients (pain: 2; fever: 1; bleeding: 1) and grade IIIb in 2 patients (bleeding). No long-term faecal incontinence was noted. CONCLUSION TEM using the SILS Port is a safe and effective procedure for local resection of benign and certain malignant tumours. Using such a cheaper device, TEM procedure could be available in any operating theatre.


title: [Endobronchial ultrasound: practical aspects and clinical applications].

text: Endobronchial ultrasound (EBUS) has become a major advance in bronchoscopy. Substantial scientific evidence has confirmed its usefulness in lung cancer diagnosis and staging, as well as in other clinical settings. It is of growing importance that endoscopists perform and interpret this imaging method accurately, in order to optimize diagnosis and treatment of their patients. The present article provides a practical and comprehensible review of the two EBUS systems currently available and its main clinical indications.


title: A new surveillance system for undiagnosed serious infectious illness for the London 2012 Olympic and Paralympic Games.

text: A new surveillance system was developed to detect possible new or emerging infections presenting as undiagnosed serious infectious illness (USII) for use during the London 2012 Olympic and Paralympic Games. Designated clinicians in sentinel adult and paediatric intensive care units (ICU/ PICUs) reported USII using an online reporting tool or provided a weekly nil notification. Reported cases were investigated for epidemiological links. A pilot study was undertaken for six months between January and July 2011 to evaluate the feasibility and acceptability of the system. In this six-month period, 5 adults and 13 children were reported by six participating units (3 ICUs, 3 PICUs). Of these 18 patients, 12 were reported within four days after admission to an ICU/PICU. Nine patients were subsequently diagnosed and were thus excluded from the surveillance. Therefore, only nine cases of USII were reported. No clustering was identified.On the basis of the pilot study, we conclude that the system is able to detect cases of USII and is feasible and acceptable to users. USII surveillance has been extended to a total of 19 sentinel units in London and the south-east of England during the London 2012 Olympic and Paralympic Games.


title: [Serological tests should be related to the aim of the testing, as well as the population].

text: Expectations are high on serological tests for SARS-CoV-2. Further knowledge of the immunity is needed, but also evaluation of the reliability of the tests. Important for the latter is for which purpose the test is conducted and how common the outcome to be identified is (antibodies). For the determination of immunity at the individual level, the specificity of the test must be very high, preferably 100%. Even tests where the specificity is perceived as high, e.g. 95% or 99%, can lead to a large proportion of false positives, if the proportion of the population actually infected is small.


title: Severe acute respiratory syndrome coronavirus infection of golden Syrian hamsters.

text: Small animal models are needed in order to evaluate the efficacy of candidate vaccines and antivirals directed against the severe acute respiratory syndrome coronavirus (SARS CoV). We investigated the ability of SARS CoV to infect 5-week-old Golden Syrian hamsters. When administered intranasally, SARS CoV replicates to high titers in the lungs and nasal turbinates. Peak replication in the lower respiratory tract was noted on day 2 postinfection (p.i.) and was cleared by day 7 p.i. Low levels of virus were present in the nasal turbinates of a few hamsters at 14 days p.i. Viral replication in epithelial cells of the respiratory tract was accompanied by cellular necrosis early in infection, followed by an inflammatory response coincident with viral clearance, focal consolidation in pulmonary tissue, and eventual pulmonary tissue repair. Despite high levels of virus replication and associated pathology in the respiratory tract, the hamsters showed no evidence of disease. Neutralizing antibodies were detected in sera at day 7 p.i., and mean titers at day 28 p.i. exceeded 1:400. Hamsters challenged with SARS CoV at day 28 p.i. were completely protected from virus replication and accompanying pathology in the respiratory tract. Comparing these data to the mouse model, SARS CoV replicates to a higher titer and for a longer duration in the respiratory tract of hamsters and is accompanied by significant pathology that is absent in mice. Viremia and extrapulmonary spread of SARS CoV to liver and spleen, which are seen in hamsters, were not detected in mice. The hamster, therefore, is superior to the mouse as a model for the evaluation of antiviral agents and candidate vaccines against SARS CoV replication.


title: [Human metapneumovirus as hospitalization cause in children under 3 years old with acute respiratory infections during 2004].

text: Human metapneumovirus was detected in 15 of 123 children (12%) younger than 3 years of age hospitalized for treatment of acute respiratory infection between July and November 2004. The virus was detected by RT-PCR directly from nasopharyngeal swabs and/or from supernatants after cell culture. Children infected with hMPV were mostly younger than one year of age (67%), all presenting with fever and cough. The main cause for hospitalization was the need for oxygen therapy (73%). Four hMPV positive children had an identifiable co-morbid condition but had a similar clinical evolution when compared to previously healthy infants. Chest radiography showed an increase in interstitial infiltrates with focal consolidation in 6 children. Obstructive bronchial syndrome and bronchiolitis, with or without pneumonia, were the most frequent diagnosis associated with hMPV positivity. A rapid and sensitive diagnostic method is required to improve diagnosis and treatment of these patients.


title: [Dose of glucocorticosteroids in the treatment of severe acute respiratory syndrome].

text: OBJECTIVE To survey the dose of glucocorticosteroids administered in patients with severe acute respiratory syndrome (SARS) and assess the effect of glucocorticosteroid doses in improving the patients' lung function. METHODS A retrospective analysis was conducted among 225 SARS patients treated in our in 2003. Oxygenation index was used as the effectness index, and the criteria for effectiveness was defiend as increase of the value of OI by 20% or above. RESULTS Glococoticostecoids were used in 59.56% of the SARS cases. The average value of OI before intravenous use of glucocorticosteroids was 237.08 mmHg, and that after the administration was 335.08 mmHg. The glucocorticosteroid doses that produce better effects were 1-3 mg/kg and 160-240 mg daily, with the total accumulative dose of 1000-2000 mg. The optimal duration of glucocorticosteroid use was 8-14 days. CONCLUSIONS For SARS treatment, Glucocorticosteroids can effectively ameliorate the SARS patients' lung symptoms and improve the lung function. The appropriate daily dose of glucocorticosteroids is 1-3 mg/kg or 160-240 mg/d for a duration of 8-14 d; the accumulative dose should be controlled around 1500 mg.


title: Complete coding sequences and phylogenetic analysis of Human Bocavirus (HBoV).

text: Human Bocavirus (HBoV) is a novel virus which can cause respiratory tract disease in infants or children. Recently, the prevalence of this virus has been studied worldwide. In this study, 18 of 252 (7.14%) nasopharyngeal suctions from infants or children between 1 month and 9 years of age with respiratory tract illness were HBoV-positive by PCR. Three positive samples were selected for sequencing the entire coding sequences using a new conserved set of primers. The results showed that the most conserved regions of HBoV are the NS1 and NP1 genes, whereas VP1 and VP2 showed frequent variations. However, the complete coding sequences showed that the variations did not depend on the origin of virus found. The complete coding sequences determined in this study can resolve the problem of an HBoV detection method, which can be advantageously implemented in laboratory detection.


title: Coronavirus HKU1 and other coronavirus infections in Hong Kong.

text: We have recently described the discovery of a novel coronavirus, coronavirus HKU1 (CoV-HKU1), associated with community-acquired pneumonia. However, the clinical spectrum of disease and the epidemiology of CoV-HKU1 infections in relation to infections with other respiratory viruses are unknown. In this 12-month prospective study, 4,181 nasopharyngeal aspirates from patients with acute respiratory tract infections were subjected to reverse transcription-PCRs specific for CoV-HKU1 and human coronaviruses NL63 (HCoV-NL63), OC43 (HCoV-OC43), and 229E (HCoV-229E). Coronaviruses were detected in 87 (2.1%) patients, with 13 (0.3%) positive for CoV-HKU1, 17 (0.4%) positive for HCoV-NL63, 53 (1.3%) positive for HCoV-OC43, and 4 (0.1%) positive for HCoV-229E. Of the 13 patients with CoV-HKU1 infections, 11 were children and 8 had underlying diseases. Similar to the case for other coronaviruses, upper respiratory infection was the most common presentation of CoV-HKU1 infections, although pneumonia, acute bronchiolitis, and asthmatic exacerbation also occurred. Despite a shorter duration of fever (mean, 1.7 days) and no difference in maximum temperature in children with CoV-HKU1 infections compared to patients with most other respiratory virus infections, a high incidence of febrile seizures (50%) was noted, which was significantly higher than those for HCoV-OC43 (14%), adenovirus (9%), human parainfluenza virus 1 (0%), and respiratory syncytial virus (8%) infections. CoV-HKU1 and HCoV-OC43 infections peaked in winter, although cases of the former also occurred in spring to early summer. This is in contrast to HCoV-NL63 infections, which mainly occurred in early summer and autumn but were absent in winter. Two genotypes of CoV-HKU1 cocirculated during the study period. Continuous studies over a longer period are warranted to ascertain the seasonal variation and relative importance of the different coronaviruses. Similar studies in other countries are required to better determine the epidemiology and genetic diversity of CoV-HKU1.


Lung and mediastinum.


title: [Study on the RNA of severe acute respiratory syndrome (SARS) associated coronavirus in the blood and excretion of convalescent patients with SARS].

text: OBJECTIVE To examine the RNA of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) in the blood and excretion of convalescent patient with SARS for prevention and treatment of the disease. METHODS A total of 276 samples, including plasma, urine, feces and sputum, obtained from 23 convalescent patients with SARS were studied at 3 time-points at least 21 days after the onset of symptoms. RNA was extracted and nested reverse transcription-polymerase chain reaction (RT-PCR) was carried out using SARS-CoV specific primers. RESULTS Among the 276 samples, SARS-CoV RNA was detected in 6 cases (38.8%) by nested RT-PCR. The positive rates of SARS-CoV RNA was 5.8% in feces and 2.9% in sputum samples but SARS-CoV RNA was not detectable in plasma and urine of all the cases. CONCLUSION The existence of SARS-CoV RNA in the excretion of some convalescent patients with SARS showed that the excretion from these patients should be carefully treated whilthe re-transmission of SARS by which, should be further studied.


title: Transgenic mice secreting coronavirus neutralizing antibodies into the milk.

text: Ten lines of transgenic mice secreting transmissible gastroenteritis coronavirus (TGEV) neutralizing recombinant monoclonal antibodies (rMAbs) into the milk were generated. The rMAb light- and heavy-chain genes were assembled by fusing the genes encoding the variable modules of the murine MAb 6A.C3, which binds an interspecies conserved coronavirus epitope essential for virus infectivity, and a constant module from a porcine myeloma with the immunoglobulin A (IgA) isotype. The chimeric antibody led to dimer formation in the presence of J chain. The neutralization specific activity of the recombinant antibody produced in transiently or stably transformed cells was 50-fold higher than that of a monomeric rMAb with the IgG1 isotype and an identical binding site. This rMAb had titers of up to 10(4) by radioimmunoassay (RIA) and neutralized virus infectivity up to 10(4)-fold. Of 23 transgenic mice, 17 integrated both light and heavy chains, and at least 10 of them transmitted both genes to the progeny, leading to 100% of animals secreting functional TGEV neutralizing antibody during lactation. Selected mice produced milk with TGEV-specific antibody titers higher than 10(6) as determined by RIA, neutralized virus infectivity by 10(6)-fold, and produced up to 6 mg of antibody per ml. Antibody expression levels were transgene copy number independent and integration site dependent. Comicroinjection of the genomic beta-lactoglobulin gene with rMAb light- and heavy-chain genes led to the generation of transgenic mice carrying the three transgenes. The highest antibody titers were produced by transgenic mice that had integrated the antibody and beta-lactoglobulin genes, although the number of transgenic animals generated does not allow a definitive conclusion on the enhancing effect of beta-lactoglobulin cointegration. This approach may lead to the generation of transgenic animals providing lactogenic immunity to their progeny against enteric pathogens.


title: Clinical experience with therapeutic dose of Low-Molecular-Weight Heparin.

text: A 71-year old gentleman with history of arterial hypertension treated with valsartan presented on was hospitalized at the Infectious Diseases Unit, University of Bologna (Italy) for severe acute respiratory syndrome- coronavirus-2 (SARS-CoV-2) and received treatment with hydroxychloroquine 200mg bid (400 mg bid the first day), azithromycin 400 mg qd, thrombotic prophylaxis with enoxaparin 4000 UI qd and Venturi mask oxygen delivering FiO2 of 31%. The case highlights the high frequency of coagulopathy in patients with moderate to severe cases of SARS-CoV-2 associated disease (COVID-19). After one week the patient significantly improved and the daily dose of enoxaparin was reduced and definitively discontinued four days later. The case highlights the high frequency of coagulopathy in patients with moderate to severe cases of SARS-CoV-2 associated disease (COVID-19). Considering the available information we believe that LMWH may represent a promising treatment for COVID-19 but further well-designed trials are needed to address these points.


title: Human immunodeficiency viral vector pseudotyped with the spike envelope of severe acute respiratory syndrome coronavirus transduces human airway epithelial cells and dendritic cells.

text: The human severe acute respiratory syndrome coronavirus (SARS-CoV) is a highly infectious virus that causes severe respiratory infections in humans. The spike envelope glycoprotein of SARS-CoV, the main determinant of SARS-CoV tropism, was isolated and used to pseudotype a human immunodeficiency virus (HIV)-based vector. Spike-pseudotyped HIV vector was generated and evaluated in vitro on well-differentiated human airway epithelial cells and bronchial explants and in vivo in murine airways. The spike envelope was less efficient at promoting HIV vector transduction of murine airway epithelium than an optimized deletion mutant of the Zaire ebolavirus envelope glycoprotein (NTD6L), which was used as a benchmark. However, spike-pseudotyped HIV vector was substantially more efficient than NTD6L-pseudotyped vector on human airway epithelium as demonstrated by lacZ gene transfer in primary cultures of epithelial cells and bronchial explants. In addition, this study shows that spike-pseudotyped HIV -based vector can efficiently transduce human dendritic cells and epithelial cells of the esophagus, which may have implications in investigating mechanisms of SARS-CoV pathogenesis. Spike-pseudotyped HIV-based vector is a novel lung-directed gene transfer vehicle that holds promise for the treatment of genetic lung diseases such as cystic fibrosis or alpha(1)-antitrypsin deficiency.


title: A new option for intracorporeal circular-stapled esophagojejunostomy in laparoscopic total gastrectomy: Roux-en-Y reconstruction with its efferent loop located at the left side of the patient to prevent twisting of the esophagojejunostomy.

text: BACKGROUND/AIMS Laparoscopic total gastrectomy (LTG) has not gained widespread acceptance because of the difficult reconstruction technique, especially for esophagojejunostomy. Although various modified procedures using a circular stapler for esophagojejunostomy have been reported, an optimal technique has not yet been established. In addition, in intracorporeal techniques, twisting of the esophagojejunostomy, which might be the cause of stenosis, is often encountered because application of the shaft is restricted. To prevent twisting of the esophagoejunostomy, we underwent LTG with Roux-en-Y reconstruction with its efferent loop located at the left side of the patient. METHODOLOGY From November 2013 to November 2014, a series of 9 patients underwent LTG with Roux-en-Y reconstruction using the transorally inserted anvil (OrVil™, Covidien, Mansfield, MA, USA), whose efferent loop was located at the left side of the patient. RESULTS No twisting of the esophagojejunostomy was encountered in all cases. In addition, no stenosis or leakage of the esophagojejunostomy occurred. CONCLUSIONS This reconstruction system may be a feasible surgical procedure in LTG.


From the founding editor.


title: Inflammatory cytokines in pediatric obstructive sleep apnea.

text: Pediatric obstructive sleep apnea (OSA) is associated with chronic systemic inflammation and with cognitive impairments. This study aimed to investigate the status of proinflammatory cytokines, particularly interleukin 17 (IL-17) and interleukin 23 (IL-23) and cognition in pediatric OSA.Controls and OSA children participated in the study. Exclusion criteria were adenotonsillectomy, heart, neurological and severe psychiatric diseases, craniofacial syndromes, and obesity. Polysomnogram was followed by serum testing for inflammatory markers and neurocognitive tests such as continuous performance task (CPT) and Wisconsin card sorting test, questionnaires, analyses of plasma high-sensitivity C-reactive protein (HS-CRP), tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1), interleukin 6 (IL-6), IL-17, and IL-23.Seventy-nine, 4 to 12-year-old subjects in 2 groups ended the study: 47 nonobese OSA children (mean age = 7.84 ± 0.56 years, body mass index [BMI] = 16.95 ± 0.47 kg/m, BMI z-score = 0.15 ± 0.21, and mean apnea-hypopnea index [AHI] = 9.13 ± 1.67 events/h) and 32 healthy control children (mean age = 7.02 ± 0.65 years, with BMI = 16.55 ± 0.58 kg/m, BMI z-score = -0.12 ± 0.27, and mean AHI = 0.41 ± 0.07 event/h) were enrolled. Serum cytokine analyses showed significantly higher levels of HS-CRP, IL-17, and IL-23 in OSA children (P = 0.002, P = 0.024, and P = 0.047). Regression test showed significant influence of HS-CRP, TNF-α, IL-6, IL-17, and specifically IL-23, with the continuous performance test and Wisconsin card sorting test.OSA children have abnormal levels of IL-17, an interleukin related to T helper 17 cells, a T helper cell involved in development of autoimmunity and inflammation. This high expression level may contribute to the complications of pediatric OSA; we also found a significant influence of inflammatory cytokines, particularly IL-23, on abnormal neurocognitive testing.


title: Transabdominal preperitoneal herniorrhaphy using laser-assisted tissue soldering in a porcine model.

text: BACKGROUND AND OBJECTIVES Collagen solder is capable of fixation of surgical meshes during laparoscopic herniorrhaphy without compromising tissue integration, increasing adhesions or inflammation. This pilot study describes development of instrumentation and techniques for transabdominal preperitoneal (TAPP) herniorrhaphy using laser-assisted soldering technology. METHODS Anesthetized 20-kg to 25-kg female Yorkshire pigs underwent laparoscopy performed using a 3-trocar technique. Peritoneal incisions were made and pockets created in the preperitoneal space for mesh placement. Parietex TEC mesh segments embedded in 60% collagen-solder were soldered to the muscle surface by using a prototype laser (1.45micro, 4.5W CW, 5mm spot, and 55 degrees C set temperature) and custom laparoscopic handpiece. Parietex TEC mesh segments (Control) were affixed to the muscle with fibrin sealant (Tisseel). Peritoneal closure was with staples (Control) or by soldering collagen embedded Vicryl mesh segments over the peritoneal incision (Mesh/TAPP). Segments were inserted using a specially designed introducer. Animals were recovered and underwent second-look laparoscopy at 6 weeks postimplantation. Mesh sites were harvested after animals were euthanized. RESULTS The mesh-solder constructs were easily inserted and affixed in the TAPP approach. Tisseel tended to drip during application, particularly in vertical and ventral locations. Postoperative healing was similar to Control segments in all cases. Mesh/TAPP closures healed without scarring or adhesion formation. DISCUSSION AND CONCLUSION Collagen-based tissue soldering permits normal wound healing and may mitigate or reduce use of staples for laparoscopic mesh fixation and peritoneal closure. Laser-assisted mesh fixation and peritoneal closure is a promising alternative for laparoscopic herniorrhaphy. Further development of this strategy is warranted.


title: First evidence of feline herpesvirus, calicivirus, parvovirus, and Ehrlichia exposure in Brazilian free-ranging felids.

text: Serum samples from 18 pumas (Puma concolor), one ocelot (Leopardus pardalis), and two little spotted cats (Leopardus tigrinus) collected from free-ranging animals in Brazil between 1998 and 2004 were tested by indirect immunofluorescence (IFA) for antibodies to feline herpesvirus 1 (FHV 1), calicivirus (FCV), coronavirus (FCoV), parvo-virus (FPV), Ehrlichia canis, Anaplasma pha-gocytophilum, and Bartonella henselae. Serum samples also were tested, by Western blot and ELISA, for feline leukemia virus (FeLV) specific antibodies and antigen, respectively, by Western blot for antibodies to feline immunodeficiency virus (FIV), and by indirect ELISA for antibodies to puma lentivirus (PLV). Antibodies to FHV 1, FCV, FCoV, FPV, FeLV, FIV, PLV or related viruses, and to B. henselae were detected. Furthermore, high-titered antibodies to E. canis or a closely related agent were detected in a puma for the first time.


Bovine intestinal spirochaetosis with dysentery.


title: [A pilot study of hydroxychloroquine in treatment of patients with moderate COVID-19].

text: OBJECTIVE To evaluate the efficacy and safety of hydroxychloroquine (HCQ) in the treatment of patients with moderate coronavirus disease 2019 (COVID-19). METHODS We prospectively enrolled 30 treatment-naïve patients with confirmed COVID-19 after informed consent at Shanghai Public Health Clinical Center. The patients were randomized 1：1 to HCQ group and the control group. Patients in HCQ group were given HCQ 400 mg per day for 5 days plus conventional treatments, while those in the control group were given conventional treatment only. The primary endpoint was negative conversion rate of SARS-CoV-2 nucleic acid in respiratory pharyngeal swab on days 7 after randomization. This study has been approved by the Ethics Committee of Shanghai Public Health Clinical Center and registered online (NCT04261517). RESULTS One patient in HCQ group developed to severe during the treatment. On day 7, nucleic acid of throat swabs was negative in 13 (86.7%) cases in the HCQ group and 14 (93.3%) cases in the control group (P>0.05). The median duration from hospitalization to virus nucleic acid negative conservation was 4 (1,9) days in HCQ group, which is comparable to that in the control group [2 (1,4) days, Z=1.27, P>0.05]. The median time for body temperature normalization in HCQ group was 1 (0,2) day after hospitalization, which was also comparable to that in the control group [1 (0,3) day]. Radiological progression was shown on CT images in 5 cases (33.3%) of the HCQ group and 7 cases (46.7%) of the control group, and all patients showed improvement in follow-up examinations. Four cases (26.7%) of the HCQ group and 3 cases (20%) of the control group had transient diarrhea and abnormal liver function (P>0.05). CONCLUSIONS The prognosis of COVID-19 moderate patients is good. Larger sample size study are needed to investigate the effects of HCQ in the treatment of COVID-19. Subsequent research should determine better endpoint and fully consider the feasibility of experiments such as sample size.


title: A preoperative hernia symptom score predicts inguinal hernia anatomy and outcomes after TEP repair.

text: BACKGROUND The Carolinas comfort scale (CCS) is an ideal tool for assessing patients’ quality-of-life post hernia repair, but its use has been barely investigated preoperatively. The aim was to quantify preoperative symptoms and assess their relevance in predicting postoperative clinical outcomes following totally extraperitoneal (TEP) inguinal hernia repair. METHODS The CCS was modified for preoperative use (modified or MCCS) by omitting mesh sensation questioning. Data collection was prospective over a 16 months period. (M)CCS questionnaires were completed preoperatively and at 2 then 6 weeks post repair. Intraoperative findings were also recorded. One hundred and four consecutive patients consented for TEP repair were included using a fibrin glue mesh fixation technique. RESULTS All three questionnaires were completed by 88 patients (84.6 %). Preoperative MCCS scores did not differ with age, obesity, the presence of bilateral or recurrent inguinal herniae or hernia type. Higher MCCS grouping [OR 4.3 (95 % CI 1.5–12.6)] and the presence of bilateral herniae [OR 8.5 (1.2–61.8)] were predictors of persisting discomfort at 6 weeks, with lower scores on MCCS [OR 16.4 (3.9–67.6), obesity (OR 9.9 91.6–63.2)] and recurrent hernia repair [OR 11.4 (1.4–91.0)] predicting increased discomfort at 2 weeks versus preoperatively. MCCS scores were inversely correlated with the size of a direct defect (r −0.42, p = 0.011) but did not differ with the intraoperative finding of an incidental femoral and/or obturator hernia. Female sex was strongly associated with recognition of a synchronous incidental hernia (5 vs 57 %, p = 0.001). CONCLUSIONS Pre- and post-operative scoring of hernia specific symptoms should be considered as part of routine surgical practice, to counsel patients on their expectations of pain and discomfort post repair and to select those who might be more appropriate for a watchful waiting approach. Females with inguinal hernia warrant complete assessment of their groin hernial orifices intraoperatively due to a high rate of synchronous incidental hernia.


title: The H1N1 influenza pandemic of 2009 in the Eastern Mediterranean Region: lessons learnt and future strategy.

text: A novel strain of influenza A virus H1N1 surfaced in Mexico in April 2009 and quickly spread across the globe, turning an epidemic into a pandemic. Within two months, the World Health Organization (WHO) declared an international health emergency and raised the threat bar from level V to level VI, i.e. containment to mitigation. During this time, the WHO Regional Office for the Eastern Mediterranean worked closely with its Member States, other stakeholders and WHO headquarters to manage the situation. This report examines the steps taken as part of this response. Programme documents were reviewed and key personnel interviewed for this study. A hallmark of the response was the establishment of the Strategic Health Operations Centre to bring together experts from different technical backgrounds at regional level. Several lessons were learnt that can provide the basis for standard operating procedures, protocols and guidelines for emergency events in future.


Out of the woods or in the eye of the storm?


Fighting 2 Enemies: Cancer and COVID-19.


title: A single amino acid change in the hemagglutinin protein of measles virus determines its ability to bind CD46 and reveals another receptor on marmoset B cells.

text: This paper provides evidence for a measles virus receptor other than CD46 on transformed marmoset and human B cells. We first showed that most tissues of marmosets are missing the SCR1 domain of CD46, which is essential for the binding of Edmonston measles virus, a laboratory strain that has been propagated in Vero monkey kidney cells. In spite of this deletion, the common marmoset was shown to be susceptible to infections by wild-type isolates of measles virus, although they did not support Edmonston measles virus production. As one would expect from these results, measles virus could not be propagated in owl monkey or marmoset kidney cell lines, but surprisingly, both a wild-type isolate (Montefiore 89) and the Edmonston laboratory strain of measles virus grew efficiently in B95-8 marmoset B cells. In addition, antibodies directed against CD46 had no effect on wild-type infections of marmoset B cells and only partially inhibited the replication of the Edmonston laboratory strain in the same cells. A direct binding assay with insect cells expressing the hemagglutinin (H) proteins of either the Edmonston or Montefiore 89 measles virus strains was used to probe the receptors on these B cells. Insect cells expressing Edmonston H but not the wild-type H bound to rodent cells with CD46 on their surface. On the other hand, both the Montefiore 89 H and Edmonston H proteins adhered to marmoset and human B cells. Most wild-type H proteins have asparagine residues at position 481 and can be converted to a CD46-binding phenotype by replacement of the residue with tyrosine. Similarly, the Edmonston H protein did not bind CD46 when its Tyr481 was converted to asparagine. However, this mutation did not affect the ability of Edmonston H to bind marmoset and human B cells. The preceding results provide evidence, through the use of a direct binding assay, that a second receptor for measles virus is present on primate B cells.


Antibody response in swine to individual transmissible gastroenteritis virus (TGEV) proteins.


title: Noninvasive ventilation in acute respiratory failure.

text: BACKGROUND Noninvasive ventilation has assumed an important role in the management of respiratory failure in critical care units, but it must be used selectively depending on the patient's diagnosis and clinical characteristics. DATA We review the strong evidence supporting the use of noninvasive ventilation for acute respiratory failure to prevent intubation in patients with chronic obstructive pulmonary disease exacerbations or acute cardiogenic pulmonary edema, and in immunocompromised patients, as well as to facilitate extubation in patients with chronic obstructive pulmonary disease who require initial intubation. Weaker evidence supports consideration of noninvasive ventilation for chronic obstructive pulmonary disease patients with postoperative or postextubation respiratory failure; patients with acute respiratory failure due to asthma exacerbations, pneumonia, acute lung injury, or acute respiratory distress syndrome; during bronchoscopy; or as a means of preoxygenation before intubation in critically ill patients with severe hypoxemia. CONCLUSION Noninvasive ventilation has assumed an important role in managing patients with acute respiratory failure. Patients should be monitored closely for signs of noninvasive ventilation failure and promptly intubated before a crisis develops. The application of noninvasive ventilation by a trained and experienced intensive care unit team, with careful patient selection, should optimize patient outcomes.


title: Emerging infectious disease issues in international adoptions: severe acute respiratory syndrome (SARS), avian influenza and measles.

text: PURPOSE OF REVIEW New emerging infections over the last few years demonstrate the potential for the introduction of epidemic illness through global migration. The increasing number of children adopted internationally (>20,000 in 2003, from the United States State Department) provides a unique situation for the spread of emerging infections through the combination of international travel by parents through areas where such infections may be contracted and the nature of the living conditions for many of the orphans being placed by this process. RECENT FINDINGS The recent literature on three emerging infections--avian influenza, severe acute respiratory syndrome (SARS) and measles--describes clinical aspects of the illnesses and their epidemiology. For avian influenza aspects of the agrarian economy in southeast Asia enabled the virus to reach the human population. The potential for further adaptation to people could set the stage for a new pandemic. SARS evolved in rural China and spread worldwide in one season with an approximate 10% mortality. Attention to public-health measures led to control of this new illness. Most recently, outbreaks of measles in Chinese orphanages have been documented. These findings demonstrate the potential of such infections to be transmitted during the process of international adoption, and in the case of measles the realization of this potential in recent reported cases from Chinese orphanages brought to the United States on commercial airlines. SUMMARY Clinicians involved in international adoption and public-health officials assessing emerging infections need to work together in monitoring these issues.


title: Effects of journey and lairage time on steers transported to slaughter in Chile.

text: Steers representative of the most common type, weight and conformation slaughtered in Chile were transported for either three or 16 hours and held in lairage for three, six, 12 or 24 hours. Measurements of liveweight, carcase weight, and the postmortem pH and colour of muscle were made to assess the economic and welfare effects of the different transport and lairage times. Compared with the short journey, the longer journey was associated with a mean (se) reduction in liveweight of 8.5 (2.8) kg, and there was a further decrease of 0.42 (0.18) kg for every hour that the animals were kept in lairage after 16 hours of transport, an increase in final muscle pH, a decrease in muscle luminosity and an increase in the proportion of carcases downgraded because they were classified as 'dark cutting The carcase weights also tended to be lower after the longer journey and after longer periods in lairage.


title: Virus-like particles as a vaccine delivery system: myths and facts.

text: Vaccines against viral disease have traditionally relied on attenuated virus strains or inactivation of infectious virus. Subunit vaccines based on viral proteins expressed in heterologous systems have been effective for some pathogens, but have often suffered from poor immunogenicity due to incorrect protein folding or modification. In this review we focus on a specific class of viral subunit vaccine that mimics the overall structure of virus particles and thus preserves the native antigenic conformation of the immunogenic proteins. These virus-like particles (VLPs) have been produced for a wide range of taxonomically and structurally distinct viruses, and have unique advantages in terms of safety and immunogenicity over previous approaches. With new VLP vaccines for papillomavirus beginning to reach the marketplace we argue that this technology has now 'come-of-age' and must be considered a viable vaccine strategy.


Anesthesia Considerations in Laryngeal Surgery.


Immune response to SARS sets up puzzling paradox.


title: The Drosophila slamdance gene: a mutation in an aminopeptidase can cause seizure, paralysis and neuronal failure.

text: We report here the characterization of slamdance (sda), a Drosophila melanogaster "bang-sensitive" (BS) paralytic mutant. This mutant exhibits hyperactive behavior and paralysis following a mechanical "bang" or electrical shock. Electrophysiological analyses have shown that this mutant is much more prone to seizure episodes than normal flies because it has a drastically lowered seizure threshold. Through genetic mapping, molecular cloning, and RNA interference, we have demonstrated that the sda phenotype can be attributed to a mutation in the Drosophila homolog of the human aminopeptidase N (APN) gene. Furthermore, using mRNA in situ hybridization and LacZ staining, we have found that the sda gene is expressed specifically in the central nervous system at particular developmental stages. Together, these results suggest that the bang sensitivity in sda mutants is caused by a defective APN gene that somehow increases seizure susceptibility. Finally, by using the sda mutation as a sensitized background, we have been able to identify a rich variety of sda enhancers and other independent BS mutations.


title: Herpes simplex virus-induced keratitis: evaluation of the role of molecular mimicry in lesion pathogenesis.

text: Viruses are suspected but usually unproven triggering factors in autoimmunity. One favored mechanism to explain the role of viruses in the genesis of autoimmunity is molecular mimicry. An immunoinflammatory blinding lesion called herpetic stromal keratitis (HSK) that follows ocular infection with herpes simplex virus (HSV) is suggested to result from a CD4(+) T-cell response to a UL6 peptide of HSV that cross-reacts with a corneal autopeptide shared with the immunoglobulin G2a(b) (IgG2a(b)) isotype. The present report reevaluates the molecular mimicry hypothesis to explain HSK pathogenesis. Our results failed to reveal cross-reactivity between the UL6 and IgG2a(b) peptides or between peptide reactive T cells and HSV antigens. More importantly, animals infected with HSV failed to develop responses that reacted with either peptide, and infection with a recombinant vaccinia UL6 vector failed to cause HSK, in spite of generating UL6 reactivity. Other lines of evidence also failed to support the molecular mimicry hypothesis, such as the failure to affect HSK severity upon tolerization of susceptible BALB/c and B-cell-deficient mice with IgG2a(b) or UL6 peptides. An additional study system revealed that HSK could be induced in mouse strains, such as the OT2 x RAG1(-/-) mice (T cell receptor transgenic recognizing OVA(323-339)) that were unable to produce CD4(+) T-cell responses to any detectable HSV antigens. Our results cast doubt on the molecular mimicry hypothesis as an explanation for the pathogenesis of HSK and indicate that if autoimmunity is involved its likely proceeds via a bystander activation mechanism.


Communicating risk: we as doctors are not alone.


title: The pathogenesis of murine coronavirus infection of the central nervous system.

text: Mouse hepatitis virus (MHV) is a positive-strand RNA virus that causes an acute encephalomyelitis that later resolves into a chronic fulminating demyelinating disease. Cytokine production, chemokine secretion, and immune cell infiltration into the central nervous system are critical to control viral replication during acute infection. Despite potent antiviral T-lymphocyte activity, sterile immunity is not achieved, and MHV chronically persists within oligodendrocytes. Continued infiltration and activation of the immune system, a result of the lingering viral antigen and RNA within oligodendrocytes, lead directly to the development of an immune-mediated demyelination that bears remarkable similarities, both clinically and histologically, to the human demyelinating disease multiple sclerosis. MHV offers a unique model system for studying host defense during acute viral infection and immune-mediated demyelination during chronic infection.


title: Ethical and legal aspects of global tobacco control.

text: On 28 February 2005, the Framework Convention on Tobacco Control came into force as a result of at least 40 countries becoming State Parties through ratification of this first ever health treaty sponsored by the World Health Organization. This article discusses the bioethical, trade, and legal aspects of global tobacco control. Special emphasis is given to globalisation of tobacco use and the challenges it poses to sovereign nations. It also advocates a bioethical basis in the pursuit of global solutions to expanding tobacco use.


title: Disparities in premature deaths from heart disease--50 States and the District of Columbia, 2001.

text: In 2001, heart disease accounted for approximately 29.0% of deaths among U.S. residents; 16.8% of those deaths occurred among persons aged <65 years. Although mortality rates from heart disease have decreased, the decline has not been uniform for all populations. One of the two overall national health objectives for 2010 is to eliminate health disparities among different segments of the U.S. population. To better understand these disparities, CDC analyzed death certificate data for premature deaths from heart disease occurring in 2001. This report summarizes the results of that analysis, which indicated that the proportion of premature heart disease deaths varied by state and was higher among blacks, American Indians/Alaska Natives (AI/ANs), Asians/Pacific Islanders (A/PIs), and Hispanics. Reducing premature death from heart disease and eliminating disparities will require preventing, detecting, treating, and controlling risk factors for heart disease in young and middle-aged adults.


title: Modulation of autoimmunity by treatment of an infectious disease.

text: Chagas' heart disease (CHD), caused by the parasite Trypanosoma cruzi, is the most common form of myocarditis in Central America and South America. Some humans and experimental animals develop both humoral and cell-mediated cardiac-specific autoimmunity during infection. Benznidazole, a trypanocidal drug, is effective at reducing parasite load and decreasing the severity of myocarditis in acutely infected patients. We hypothesized that the magnitude of autoimmunity that develops following T. cruzi infection is directly proportional to the amount of damage caused by the parasite. To test this hypothesis, we used benznidazole to reduce the number of parasites in an experimental model of CHD and determined whether this treatment altered the autoimmune response. Infection of A/J mice with the Brazil strain of T. cruzi leads to the development of severe inflammation, fibrosis, necrosis, and parasitosis in the heart accompanied by vigorous cardiac myosin-specific delayed-type hypersensitivity (DTH) and antibody production at 21 days postinfection. Mice succumbed to infection within a month if left untreated. Treatment of infected mice with benznidazole eliminated mortality and decreased disease severity. Treatment also reduced cardiac myosin-specific DTH and antibody production. Reinfection of treated mice with a heart-derived, virulent strain of T. cruzi or immunization with myosin led to the redevelopment of myosin-specific autoimmune responses and inflammation. These results provide a direct link between the levels of T. cruzi and the presence of autoimmunity and suggest that elimination of the parasite may result in the reduction or elimination of autoimmunity in the chronic phase of infection.


title: Detailed study of graft sinking and worsening of the fused segment angle in patients with cervical disease treated with the Williams-Isu method.

text: Detailed changes involved in the worsening of the fused segment angle were assessed after application of the Williams-Isu method using autologous bone grafts from cervical vertebral bodies in 30 patients with cervical disease treated by single-level anterior fusion. The mean follow-up duration was 25.4 months. The fused segment angle was measured on serial radiographs. Whole cervical spine alignment changed from 12.8° to 9.9°. The alignment of the fused segment worsened by mean 3.3°. To elucidate the characteristics of worsening of the fused segment, the 30 patients were divided into 2 groups: Group I (n = 20) without and Group II (n = 10) with postoperative worsening of the fused segment. The loss in the fused segment angle was significantly greater in Group II (8.0°) than Group I (0.9°). Preoperative range of motion and disc height were significantly greater in Group II than Group I. Worsening of the fused segment angle occurred within 1 month in Group I, whereas stabilization was observed after 3 months in Group II. Graft subsidence was primarily posterior and inferior. Our results indicate that the preoperative range of motion and disc height of the fused segment must be considered to prevent worsening in that segment after anterior fusion. Such detailed information is useful for the selection and postoperative monitoring of patients eligible for treatment by the Williams-Isu method.


title: Lymphoid organ alterations enhanced by sub-lethal doses of coronaviruses in experimentally induced Trypanosoma cruzi infection in mice.

text: The effect of sub-lethal doses of coronaviruses on the course of disease in CBA mice experimentally infected with a mildly pathogenic strain of Trypanosoma cruzi was investigated. Mice were inoculated with either T. cruzi, 0.1 median lethal dose (LD50) of coronavirus (mouse hepatitis virus [MHV-3] or virus X), or both pathogens. Levels of parasitemia, mortality, and the extent of pathologic alterations in lymphoid organs were determined. Mice inoculated with T. cruzi had mild alterations in their lymphoid organs and survived infection. In contrast, mice inoculated with both pathogens died, and had significantly higher levels of parasitemia and profound alterations in lymphoid organs. These results indicate that the pathologic profile of T. cruzi infection can be profoundly altered by subclinical infection with coronaviruses.


title: High prevalence of turkey parvovirus in turkey flocks from Hungary experiencing enteric disease syndromes.

text: Samples collected in 2008 and 2009, from 49 turkey flocks of 6 to 43 days in age and presenting clinical signs of enteric disease and high mortality, were tested by polymerase chain reaction and reverse transcription-polymerase chain reaction for the presence of viruses currently associated with enteric disease (ED) syndromes: astrovirus, reovirus, rotavirus, coronavirus, adenovirus, and parvovirus. Turkey astroviruses were found in 83.67% of the cases and turkey astrovirus 2 (TAst-2) in 26.53%. The investigations directly demonstrated the high prevalence of turkey parvovirus (TuPV) in 23 flocks (46.9%) experiencing signs of ED, making this pathogen the second most identified after astroviruses. Phylogenetic analysis on a 527 base pair-long region from the NS1 gene revealed two main clusters, a chicken parvovirus (ChPV) and a TuPV group, but also the presence of a divergent branch of tentatively named "TuPV-like ChPV" strains. The 23 Hungarian TuPV strains were separately positioned in two groups from the American origin sequences in the TuPV cluster. An Avail-based restriction fragment length polymorphism assay has also been developed for the quick differentiation of TuPV, ChPV, and divergent TuPV-like ChPV strains. As most detected enteric viruses have been directly demonstrated in healthy turkey flocks as well, the epidemiology of this disease complex remains unclear, suggesting that a certain combination of pathogens, environmental factors, or both are necessary for the development of clinical signs.


title: Immunological features in patients with pneumonitis due to influenza A H1N1 infection.

text: BACKGROUND Pneumonitis induced by pandemic influenza A H1N1 has a potential to cause respiratory failure, which is a risk factor for death. The underlying immunopathological mechanisms, however, have not yet been fully elucidated. PATIENTS AND METHODS We investigated changes in plasma cytokines, T cell subsets, and C-reactive protein (CRP) in 16 hospitalized patients with pneumonia caused by 2009 H1N1 influenza infection. The patients were classified into a severe disease group and a mild disease group according to PaO2. RESULTS Cytokine profiles showed no changes in interferon gamma (IFN-gamma), interleukin 6 (IL-6), IL-8, or transforming necrosis factor alpha (TNF-alpha) levels throughout the observation period. Transforming growth factor beta (TGF-beta1) was overproduced in the severe group but not in the mild group. Accordingly, we also found some signs of pulmonary fibrosis during the recovery period. Elevated CRP levels and lymphopenia were common in both the severe and the mild group. After treatment, there was a significant elevation in lymphocytes in both groups, but a significant decrease in CRP in the mild group. Lymphocyte counts and CRP levels rapidly recovered to normal levels in all survivors posttreatment; otherwise it seemed to be related to poor prognosis. CONCLUSIONS Serial measurements of cytokines showed that only TGF-beta1 was overproduced, possibly in relation to the early use of corticosteroids, which may have downregulated immune responses to H1N1 infection. Pretreatment TGF-beta1 plasma concentrations and absolute lymphocyte counts were independent predictors of severity. However, the role of elevated TGF-beta1 in H1N1 infection-associated pulmonary fibrosis requires further investigation.


title: Impaired angiogenesis in aminopeptidase N-null mice.

text: Aminopeptidase N (APN, CD13; EC 3.4.11.2) is a transmembrane metalloprotease with several functions, depending on the cell type and tissue environment. In tumor vasculature, APN is overexpressed in the endothelium and promotes angiogenesis. However, there have been no reports of in vivo inactivation of the APN gene to validate these findings. Here we evaluated, by targeted disruption of the APN gene, whether APN participates in blood vessel formation and function under normal conditions. Surprisingly, APN-null mice developed with no gross or histological abnormalities. Standard neurological, cardiovascular, metabolic, locomotor, and hematological studies revealed no alterations. Nonetheless, in oxygen-induced retinopathy experiments, APN-deficient mice had a marked and dose-dependent deficiency of the expected retinal neovascularization. Moreover, gelfoams embedded with growth factors failed to induce functional blood vessel formation in APN-null mice. These findings establish that APN-null mice develop normally without physiological alterations and can undergo physiological angiogenesis but show a severely impaired angiogenic response under pathological conditions. Finally, in addition to vascular biology research, APN-null mice may be useful reagents in other medical fields such as malignant, cardiovascular, immunological, or infectious diseases.