title: Genomic Analysis of Viral Outbreaks.

text: Genomic analysis is a powerful tool for understanding viral disease outbreaks. Sequencing of viral samples is now easier and cheaper than ever before and can supplement epidemiological methods by providing nucleotide-level resolution of outbreak-causing pathogens. In this review, we describe methods used to answer crucial questions about outbreaks, such as how they began and how a disease is transmitted. More specifically, we explain current techniques for viral sequencing, phylogenetic analysis, transmission reconstruction, and evolutionary investigation of viral pathogens. By detailing the ways in which genomic data can help us understand viral disease outbreaks, we aim to provide a resource that will facilitate the response to future outbreaks.


title: Functional expression of endothelial nitric oxide synthase fused to green fluorescent protein in transgenic mice.

text: The activity of endothelial nitric oxide synthase (eNOS) is subject to complex transcriptional and post-translational regulation including the association with several proteins and variations in subcellular distribution. In the present study we describe a transgenic mouse model expressing eNOS fused to green fluorescent protein (GFP), which allows the study of localization and regulation of eNOS expression. We tested the functionality of eNOS in the eNOS-GFP mice. Expression of eNOS was restricted to the endothelial lining of blood vessels in various tissues tested, without appreciable expression in non-endothelial cells. Activity of the enzyme was confirmed by assaying the conversion of L-arginine to L-citrulline. NO production in isolated vessels was increased in transgenic mice when compared to non-transgenic control animals (4.88 +/- 0.59 and 2.48 +/- 0.47 micro mol/L NO, respectively, P < 0.005). Both the mean aortic pressure and the pulmonary artery pressure were reduced in eNOS-GFP mice (both approximately 30%, P < 0.05). Plasma cholesterol levels were also slightly reduced ( approximately 20%, P < 0.05). In conclusion, eNOS-GFP mice express functional eNOS and provide a unique model to study regulation of eNOS activity or eNOS-mediated vascular events, including response to ischemia, response to differences in shear stress, angiogenesis and vasculogenesis, and to study the subcellular distribution in relation with functional responses to these events.


title: Comparison of immunoglobulin G responses to the spike and nucleocapsid proteins of severe acute respiratory syndrome (SARS) coronavirus in patients with SARS.

text: Both the nucleocapsid (N) and the spike (S) proteins of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) are able to induce strong humoral responses in humans following an infection. To compare the immunoglobulin G (IgG) responses to the S and N proteins of SARS-CoV in SARS patients during the manifestation/convalescent period with those during the postinfection period, serum samples were collected from hospitalized SARS patients within 6 weeks after the onset of illness (set 1; 57 sequential samples from 19 patients) or 2 to 3 months after their recovery (set 2; 33 postinfection samples from 33 subjects). Serum samples from 100 healthy blood donors (set 3), collected in 2002, were also included. The specific IgG response to whole virus, the fragment from positions 450 to 650 of the S protein (S450-650), and the full-length N protein of SARS-CoV were measured by enzyme-linked immunosorbent assays (ELISAs). Western blot assays were carried out to confirm the ELISA results. Fifty-one of the serum samples in set 1 (89%) bound to the N protein, a proportion similar to that which recognized whole virus (79%) and the S-protein fragment (77%). All 33 serum samples from set 2 were strongly positive for N-protein-specific IgG, while 27 (82%) were positive for anti-S450-650 IgG. Two of the serum samples from set 3 were strongly positive for anti-N-protein IgG but not anti-S450-650 IgG. Similar levels of IgG responses to the S and N proteins were observed in SARS patients during the manifestation and convalescent stages. In the postinfection period, however, a number of patients had much lower serum IgG levels against S450-650 than against the N protein.


title: On the hybridization of memetic algorithms with branch-and-bound techniques.

text: Branch-and-bound (BnB) and memetic algorithms represent two very different approaches for tackling combinatorial optimization problems. However, these approaches are compatible. In this correspondence, a hybrid model that combines these two techniques is considered. To be precise, it is based on the interleaved execution of both approaches. Since the requirements of time and memory in BnB techniques are generally conflicting, a truncated exact search, namely, beam search, has opted to be carried out. Therefore, the resulting hybrid algorithm has a heuristic nature. The multidimensional 0-1 knapsack problem and the shortest common supersequence problem have been chosen as benchmarks. As will be shown, the hybrid algorithm can produce better results in both problems at the same computational cost, especially for large problem instances.


docid: zfcdpacg

title: [Clinical analysis of multiple organ dysfunction syndrome in patients suffering from SARS].

text: OBJECTIVE To analyze the features of organ dysfunction in severe acute respiratory syndrome (SARS) and the factors inducing multiple organ dysfunction syndrome (MODS) in these patients. METHODS The clinical data of 135 SARS patients and 13 patients with severe SARS complicated by MODS were studied retrospectively. RESULTS SARS could result in organ dysfunction. Its incidence rate was high, the patients' condition was severe, and the mortality was higher. SARS could end in MODS and the mortality was 92.3x. The main dysfunctioning organs were the lung (76.9x), the immune system (92.3x), the cardiovascular system (30.8x), the blood system (61.5x), the liver (38.5x), and the kidney (53.8x). CONCLUSION Dysfunction of organs are closely correlated with prognosis of SARS. The causes of MODS in SARS are probably injury to the immune system and systemic inflammatory response syndrome.


title: A new application of sequence Fourier analysis to specific ligand-protein interactions.

text: A frequency (or sequence)-dependent rule seems to be conserved in a simple and concerted interaction of various human proteins.


title: Prioritization and willingness to pay for bariatric surgery: the patient perspective.

text: BACKGROUND Access to publicly funded bariatric surgery is limited, potential candidates face lengthy waits, and no universally accepted prioritization criteria exist. We examined patients' perspectives regarding prioritization for surgery. METHODS We surveyed consecutively recruited patients awaiting bariatric surgery about 9 hypothetical scenarios describing patients waiting for surgery. Respondents were asked to rank the priority of these hypothetical patients on the wait list relative to their own. Scenarios examined variations in age, clinical severity, functional impairment, social dependence and socioeconomic status. Willingness to pay for faster access was assessed using a 5-point ordinal scale and analyzed using multivariable logistic regression. RESULTS The 99 respondents had mean age of 44.7 ± 9.9 years, 76% were women, and the mean body mass index was 47.3 ± SD 7.6. The mean wait for surgery was 34.4 ± 9.4 months. Respondents assigned similar priority to hypothetical patients with characteristics identical to theirs (p = 0.22) and higher priority (greater urgency) to those exhibiting greater clinical severity (p < 0.001) and functional impairment (p = 0.003). Lower priority was assigned to patients at the extremes of age (p = 0.006), on social assistance (p < 0.001) and of high socioeconomic status (p < 0.001). Most (85%) respondents disagreed with payment to expedite access, although participants earning more than $80 000/year were less likely to disagree. CONCLUSION Most patients waiting for bariatric surgery consider greater clinical severity and functional impairments related to obesity to be important prioritization indicators and disagreed with paying for faster access. These findings may help inform future efforts to develop acceptable prioritization strategies for publicly funded bariatric surgery.


title: First epidemiological study of sars.

text: Health authorities worldwide, especially in Asia and the Pacific region, are seeking effective public health interventions to tackle the continuing epidemic of severe acute respiratory syndrome (SARS).


title: Bioprospecting for microbial endophytes and their natural products.

text: Endophytic microorganisms are to be found in virtually every plant on earth. These organisms reside in the living tissues of the host plant and do so in a variety of relationships, ranging from symbiotic to slightly pathogenic. Because of what appears to be their contribution to the host plant, the endophytes may produce a plethora of substances of potential use to modern medicine, agriculture, and industry. Novel antibiotics, antimycotics, immunosuppressants, and anticancer compounds are only a few examples of what has been found after the isolation, culture, purification, and characterization of some choice endophytes in the recent past. The potential prospects of finding new drugs that may be effective candidates for treating newly developing diseases in humans, plants, and animals are great.


title: Antigen delivery systems.

text: Many vaccine candidates are highly purified, sometimes monomeric antigens and as a result, not very immunogenic. Antigen delivery systems optimize the presentation of antigens. They also play a major role in solving the problem of there being an increasing number of vaccines but limited opportunities in which to include these vaccines in immunization programs. The number of injections is restricted and combining vaccines may lead to immunological and physicochemical incompatibility. In this review, the current status with respect to parenteral and mucosal delivery systems is discussed. These include lipid-based systems such as liposomes and immunostimulating complexes, as well as polymeric microspheres. In addition, developments in needle-free, dermal delivery devices such as jet injectors, microneedles and patches are presented.


title: Transfer of endoplasmic reticulum and Golgi retention signals to human immunodeficiency virus type 1 gp160 inhibits intracellular transport and proteolytic processing of viral glycoprotein but does not influence the cellular site of virus particle budding.

text: In this study, specific signals known to mediate endoplasmic reticulum or Golgi localization of transmembrane proteins have been transferred to the human immunodeficiency virus type 1 (HIV-1) env gene product. The intracellularly retained recombinant glycoproteins were not proteolytically processed to gp120 and gp41, which is further evidence that this process occurs at a later stage in the transport pathway, presumably within or near the trans-Golgi network. Since the subcellular localization of the viral glycoproteins of enveloped viruses can be one of the factors determining the cellular site of particle assembly and release, experiments were performed to determine if this property was altered by coexpression of the recombinant HIV-1 glycoproteins. When wild-type virus was compared to mutant virus encoding the intracellularly retained glycoproteins, the extent of HIV-1 particle release into the extracellular medium remained unaffected, and electron-microscopic analysis did not reveal any significant alteration in the cellular sites of particle assembly and budding. Thus, in COS-7 cells, altered subcellular localization of the viral glycoprotein does not exert a dominant influence on the assembly site of the HIV-1 particle.


docid: bcwfagau

title: Small intestinal strangulation caused by a mesodiverticular band and diverticulum on the mesenteric border of the small intestine in a horse.

text: A 4-year-old Quarter horse stallion was presented for acute abdominal pain. Exploratory celiotomy revealed a mesenteric diverticulum of the jejunum and mesodiverticular band that were associated with small intestinal strangulation. Resection and anastomosis were performed. A second celiotomy was performed due to adhesions. The horse recovered completely and returned to training.


The protective effect of childhood infections.


title: Hepatobiliary pathology.

text: PURPOSE OF REVIEW This review highlights recent publications on hepatobiliary pathology concerning several unusual types of hepatitis, fatty liver disease, disorders of the biliary tree and other topics that have a substantial impact on liver biopsy interpretation. RECENT FINDINGS In the outbreak of severe acute respiratory syndrome (SARS), many patients had abnormalities in liver function tests. Liver biopsy findings in three cases were reported that showed a generic picture of hepatitis, with exceptionally increased mitotic activity. The role of portal myofibroblasts in cirrhosis was examined in several studies. A newly described lesion, isolated ductular hyperplasia (IDH) was found in patients with prolonged abnormalities of liver function tests of uncertain origin. Hyperplastic, well-differentiated bile ductules were seen on liver biopsy in the absence of any identifiable biliary disease. Hereditary hemochromatosis is now a complex entity with various clinicopathological forms based on mutations in the HFE gene and other iron-homeostatic genes such as transferrin receptor 2 and ferroportin 1. In some of these heritable forms of primary iron overload, stainable iron is present in both hepatocytes and Kupffer cells. After liver transplantation, differentiating recurrent HCV infection from acute rejection on liver biopsy is problematic, with exceptionally low inter- and intra-observer reliability shown in one study. SUMMARY The hepatitis associated with the SARS coronavirus, Isolated Ductular Hyperplasia in patients with liver function test abnormalities and other topics with pathologic relevance are reviewed.


Prevention of calf diarrhea with an immunoglobulin diet in beef herds.


title: Opportunities and limitations of SELDI-TOF-MS in biomedical research: practical advices.

text: Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, or surface-enhanced laser desorption/ionization ProteinChip technology, has been widely used in obtaining the quantitative profiles of tissue proteomes, particularly plasma proteomes. Its high-throughput nature and simplicity in its experimental procedures have allowed this technology to become a popular research tool for biomarker discovery in the past 5 years. After accumulating more research experiences, researchers now have a better understanding of the characteristics and limitations of this technology, as well as the pitfalls in biomarker research, by undertaking a comparative proteomic approach. This review provides an overview of the surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, discusses its limitations and provides some possible solutions to help apply this technology to biomarker research.


title: PCR for detection of rubella virus RNA in clinical samples.

text: A reverse transcription nested PCR (RT-PCR) assay for the detection of rubella virus RNA using primers from the E1 open reading frame was established. This assay was found to be sensitive (detecting approximately two synthetic RNA copies and RNA extracted from 0.1 50% tissue culture infective dose of rubella virus) and specific; five wild-type rubella strains and four vaccine strains were detected, and no nonspecific amplification of 16 other RNA viruses or RNAs from seven cell types occurred. Rubella virus RNA was detected in 12 pharyngeal swabs from patients with serologically confirmed rubella; these RT-PCR results were in complete agreement with virus isolation. Analysis of products of conception obtained after confirmed primary maternal rubella infection by RT-PCR gave 92% agreement (12 of 13 samples) with virus isolation. No false-positive results were obtained. The potential use of this assay for prenatal diagnosis of congenital rubella infection and for investigating aspects of the pathogenesis of chronic disease is discussed.


title: Transanal endoscopic microsurgery.

text: Transanal endoscopic microsurgery, although technically challenging, offers a viable and perhaps superior outcome to radical abdominal or traditional transanal excision of rectal neoplasia. Appropriate training and case selection, as with any new technology, are mandatory to help ensure optimal results.


Changes in monocyte counts and expression of mCD(14) and HLA-DR in the peripheral blood of patients with severe acute respiratory syndrome.


title: Maternally derived recombinant human anti-hantavirus monoclonal antibodies are transferred to mouse offspring during lactation and neutralize virus in vitro.

text: Transgenic mice expressing a recombinant human monoclonal antibody (rHMAb) against hantavirus were generated. These mice could be used as models to explore the possibilities of producing rHMAbs for therapeutic purposes. The highest concentration of the rHMAb in the milk of the transgenic females was 6.6 mg/ml. The rHMAb was also detected in the sera of pups fed by the transgenic females. Both the rHMAbs in the milk of transgenic mice and those in the sera of suckling pups were found to be active against hantaviruses, although the light chain of the antibody absorbed by the pups was modified by N-linked glycosylation.


title: Nephropathogenesis of chickens experimentally infected with various strains of infectious bronchitis virus.

text: Four-day-old specific-pathogen-free chickens were inoculated by eyedrop with four different strains (Gray, JMK, CV56b, and Wolgemuth) of infectious bronchitis virus (IBV). Birds were monitored clinically and euthanatized at 1, 4, 7, and 14 days postinfection and tissues were collected for virus isolation, histopathologic examination, in situ hybridization (ISH), and immunohistochemistry (IHC). Clinical disease was severe in chickens infected with Wolgemuth, but no overt disease was observed with the other strains. Virus was isolated from the kidneys of chickens infected with the Gray-, CV56b-, and Wolgemuth-strains of IBV. Histologically, interstitial nephritis was evident in chickens infected with these same 3 strains. However, viral nucleic acid and antigen were detected only with Wolgemuth-infected kidneys by ISH and IHC. These results indicate that the pathological changes in kidneys from chickens infected with Gray and CV56b may not have resulted from the cytolytic action of the virus.


title: Multi-phase approach to eradicate enzootic mouse coronavirus infection.

text: Infections with mouse coronavirus (also known as mouse hepatitis virus, MHV) are common and prompt concern because the adverse research effects of infection have been well documented. The animal facility we describe had contained an enzootic infection of mouse coronavirus for more than a decade. Eradication of the virus had been tried with limited success in the past. With an increase in the populations of immune-compromised and transgenic animals elsewhere in the facility, eradication of the virus became a high priority. The affected animals were in multiple breeding colonies comprising more than 4000 mice representing more than 50 site-specific strains, lines, and sublines. A task force designed to ensure representation of all interested parties developed a multi-phase approach which included (a) culling of nonessential animals, (b) cessation of breeding, (c) testing and culling of individual animals, and (d) cross-fostering to clean dams. To date, all of the strains, lines, and sublines manipulated through this multi-phase eradication approach continue to test negative for mouse coronavirus. This multi-phase approach may be useful for other institutions attempting to eradicate mouse coronavirus.


title: Barbed Sutures in Body-Contouring: Outcome Analysis of 695 Procedures in 623 Patients and Technical Advances.

text: INTRODUCTION Surgical wound closure is often complicated by suture-related issues. The recent introduction of knotless barbed sutures may address the shortcomings of conventional sutures and offer the additional benefit of reduced operating time. In this paper, we describe our experience with barbed sutures for body-contouring procedures. We share technical insights and evaluate postoperative complications. PATIENTS AND METHODS A retrospective assessment of prospectively collected data over a period of more than 5 years was undertaken. Six hundred twenty-three consecutive patients underwent 695 body-contouring procedures with barbed suture closure. Patients were followed for at least 12 weeks postoperatively. Patient demographics, operation time as well as suture-related complications, such as wound dehiscence and wound site infection were recorded and analyzed. RESULTS Barbed sutures can facilitate skin closure, rectus plication, quilting, and deep layer closure in body-contouring procedures. The average operating time in our study cohort was 108 min with reduction mammoplasty being the quickest (94 min) and bodylift (156 min) being the slowest procedure. Sixty-eight patients experienced suture-related complications resulting in an overall complication rate of 9.7 % with thigh lift having the most (15 %) and reduction mammoplasty (7 %) the fewest adverse events. CONCLUSION The use of barbed sutures allows quick closure of lengthy body-contouring incision lines with low complication rates. Our observations support that barbed sutures are safe, convenient and effective. In our hands barbed sutures appear to be superior to traditional wound-closure techniques in body-contouring procedures. LEVEL OF EVIDENCE IV This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


title: Ethical considerations for emergency care providers during pandemic influenza--ready or not...

text: When an infectious pandemic occurs in the United States, emergency care providers (ECPs) will be on the frontlines caring for infected, potentially infected, and non-infected patients. Logistically, the current emergency care system is not ready for a pandemic, but are the providers ethically ready? Some of the most difficult and challenging issues that will be raised during a pandemic will be ethical in nature. An ECP likely will be confronted with ethical values and value conflicts underlying restriction of liberty, duty to care, and resource allocation. This report summarizes the ethical concerns and challenges that ECPs face during an infectious pandemic, and raises ethical questions that may arise related to the role of an ECP as a healthcare provider and stakeholder.


Assay to detect H5N1 oseltamivir resistance.


Penetrating Spinal Cord Trauma: A Unique Patient Presentation.


docid: kdpwadgf

title: Combined effect of dark and wounding on regeneration potential of Houttuynia cordata Thunb. leaves.

text: Houttuynia cordata is one of the most potential medicinal and edible wild herb whose resources have decreased sharply due to excessive exploitation. Besides its slow agamic propagation, problems of browning and non-dedifferentiation have prevented the application of micropropagation in H. cordata. Through 4 week pre-culture in darkness and wounding after 1 week pre-culture, the browning rate of leaf explants decreased significantly and resulted in efficient regeneration (20.64 +/- 5.94 adventitious buds per explant) on the induction medium. The protocol shall facilitate conservation and commercial cultivation of the endangered species.


docid: shqavwpf

title: Concept of operations for triage of mechanical ventilation in an epidemic.

text: The recent outbreak of severe acute respiratory syndrome and the growing potential of an influenza pandemic force us to consider the fact that despite great advances in critical care medicine, we lack the capacity to provide intensive care to the large number of patients that may be generated in an epidemic or multisite bioterrorism event. Because many epidemic and bioterrorist agent illnesses involve respiratory failure, mechanical ventilation is a frequently required intervention but one that is in limited supply. In advance of such an event, we must develop triage criteria that depend on clinical indicators of survivability and resource utilization to allocate scarce health care resources to those who are most likely to benefit. These criteria must be tiered, flexible, and implemented regionally, rather than institutionally, with the backing of public health agencies and relief of liability. This report provides a sample concept of operations for triage of mechanical ventilation in epidemic situations and discusses some of the ethical principles and pitfalls of such systems.


A place in our hearts for the lowly angiotensin 1-7 peptide?


title: Contrast-enhanced computed tomography angiography and volume-rendered imaging for evaluation of cellophane banding in a dog with extrahepatic portosystemic shunt.

text: A 4-year-old, 1.8 kg, male, castrated Maltese was presented for evaluation of urolithiasis. Urinary calculi were composed of ammonium biurate. Preprandial and postprandial bile acids were 44.2 and 187.3 micromol/l, respectively (reference ranges 0-10 and 0-20 micromol/l, respectively). Single-phase contrast-enhanced computed tomography angiography (CTA) with volume-rendered imaging (VRI) was obtained. VRI revealed a portocaval shunt originating just cranial to a tributary of the gastroduodenal vein and draining into the caudal vena cava at the level of the epiploic foramen. CTA revealed a 3.66 mm-diameter shunt measured at the level of the termination of the shunt and a 3.79 mm-diameter portal vein measured at the level between the origin of the shunt and the porta of the liver. Surgery was performed using cellophane banding without attenuation. Follow-up single-phase CTA with VRI was obtained 10 weeks after surgery. VRI revealed no evidence of portosystemic communication on the level of a cellophane band and caudal to the cellophane band. CTA demonstrated an increased portal vein diameter (3.79-5.27 mm) measured at the level between the origin of the shunt and the porta of the liver. Preprandial and postprandial bile acids were 25 and 12.5 micromol/l, respectively (aforementioned respective reference ranges), 3 months post-surgery. No problems were evident at 6 months.


title: The experience of quarantine for individuals affected by SARS in Toronto.

text: OBJECTIVE The purpose of this study was to explore the experience of home quarantine during the severe acute respiratory syndrome (SARS) outbreak in Toronto in 2003. DESIGN Qualitative descriptive design. SAMPLE Stratified random sampling techniques were used to generate a list of potential participants, who varied in terms of gender and closeness of exposure to someone with suspected SARS (contact level). Twenty-one individuals participated in the study. MEASUREMENTS All interviews were audiotaped and followed a semistructured interview guide. Participants were invited to describe their experience of quarantine in detail including their advice for Public Health. RESULTS The experience followed a trajectory of stages beginning before quarantine and ending after quarantine. Despite individual differences, common themes of uncertainty, isolation, and coping intersected the data. CONCLUSIONS Public Health has a dual role of monitoring compliance and providing support to people in quarantine. This study has implications for public health policy and practice in planning for future public health emergencies in terms of the information and the resources required to mount an effective response.


title: [Study of infusion of oxygen-enriched liquid to correct severe hypoxemia in infectious diseases: a report of pilot clinical study].

text: OBJECTIVE To investigate a new therapy for effectively correcting severe hypoxemia in patients with infectious diseases by infusion of oxygen-enriched liquid, in order to raise the partial pressure of blood oxygen without passing through pathologically damaged alveoli of such patients. METHODS Intravenous drip with oxygen-enriched liquids was given to 6 cases suffering from severe acute respiratory syndrome (SARS), and 3 cases of acquired immune deficiency syndrome (AIDS) in the course of treatment for 1 to 5 days, 500-700 ml per day. RESULTS For all the 9 SARS cases, their hypoxemia was gradually corrected to normal in 20 minutes' or 4 hours' intravenous drip with oxygen-enriched liquid. Respiratory rate decreased from 29-49 breath/min to 18-22 breath/min, heart rate decreased from 89-145 beats/min to 60-79 beats/min, two faint patients regained consciousness, hypoxemia was redressed, partial pressure of oxygen in artery increased from 56 mm Hg (1 mm Hg=0.133 kPa) to 87 mm Hg, saturation of oxygen increased from 0.89 to 0.96. CONCLUSION Intravenous drip of the oxygen-enriched liquid effectively helped correct the hypoxemia of SARS and other infectious diseases cases by bypassing the diseased alveoli through which oxygen would not pass into the blood by conventional oxygen inhalation. This therapy of oxygen-enriched liquid infusion could be quite life-saving in the combined treatment for SARS and other infectious diseases.


title: Haematological manifestations in patients with severe acute respiratory syndrome: retrospective analysis.

text: OBJECTIVES To evaluate the haematological findings of patients with severe acute respiratory syndrome (SARS). DESIGN Analysis of the demographic, clinical, and laboratory characteristics of patients with SARS. SETTING Prince of Wales Hospital, Hong Kong. Subjects All patients with a diagnosis of SARS between 11 March and 29 March 2003 who had no pre-existing haematological disorders. MAIN OUTCOME MEASURES Clinical end points included the need for intensive care and death. Univariate and multivariate analyses were performed to examine factors associated with adverse outcome. RESULTS 64 male and 93 female patients were included in this study. The most common findings included lymphopenia in 153 (98%) of the 157 patients, neutrophilia in 129 (82%), thrombocytopenia in 87 patients (55%), followed by thrombocytosis in 77 (49%), and isolated prolonged activated partial thromboplastin time in 96 patients (63%). The haemoglobin count dropped by more than 20 g/l from baseline in 95 (61%) patients. Four patients (2.5%) developed disseminated intravascular coagulation. Lymphopenia was shown in haemato-lymphoid organs at postmortem examination. Multivariate analysis showed that advanced age and a high concentration of lactate dehydrogenase at presentation were independent predictors of an adverse outcome. Subsets of peripheral blood lymphocytes were analysed in 31 patients. The counts of CD4 positive and CD8 positive T cells fell early in the course of illness. Low counts of CD4 and CD8 cells at presentation were associated with adverse outcomes. CONCLUSIONS Abnormal haematological variables were common among patients with SARS. Lymphopenia and the depletion of T lymphocyte subsets may be associated with disease activity.


title: Type IV hypersensitivity in the pathogenesis of FIPV-induced lesions.

text: In focal lesions of feline infectious peritonitis (FIP), the cells involved in the delayed-type hypersensitivity were identified in formalin-fixed paraffin-embedded and frozen samples taken from 35 affected cats. The clinical diagnosis of FIP was confirmed by necropsy, histology and direct immunofluorescence against the coronaviruses on cryostatic sections. The immune cells were detected immunohistochemically by the Avidin-Biotin-Complex (ABC) method using either polyclonal antibodies against lymphoid antigens (CD3) or monoclonal antibodies against lymphoid (PAN-T, CD4, CD8) and myeloid antigens (MAC387). Better identification of T cells and macrophages was found on formalin-fixed paraffin-embedded sections than on cryostatic ones, while T lymphocyte subpopulations could be differentiated only in cryostatic sections. Type IV hypersensitivity was detected in focal feline infectious peritonitis virus (FIPV)-induced lesions from progressive activation of T lymphocytes, mainly CD4+, and the presence of granulocytes and macrophages. The FIPV-induced lesions could be studied as examples of granulomas caused by unconventional antigens, such as viruses or immune complexes.


title: Evaluation of syndromic surveillance in the Netherlands: its added value and recommendations for implementation.

text: In the last decade, syndromic surveillance has increasingly been used worldwide for detecting increases or outbreaks of infectious diseases that might be missed by surveillance based on laboratory diagnoses and notifications by clinicians alone. There is, however, an ongoing debate about the feasibility of syndromic surveillance and its potential added value. Here we present our perspective on syndromic surveillance, based on the results of a retrospective analysis of syndromic data from six Dutch healthcare registries, covering 1999–2009 or part of this period. These registries had been designed for other purposes, but were evaluated for their potential use in signalling infectious disease dynamics and outbreaks. Our results show that syndromic surveillance clearly has added value in revealing the blind spots of traditional surveillance, in particular by detecting unusual, local outbreaks independently of diagnoses of specific pathogens, and by monitoring disease burden and virulence shifts of common pathogens. Therefore we recommend the use of syndromic surveillance for these applications.


Urgent need for empirical research into whaling and whale watching.


title: [Lung Cancer and Respiratory Infections].

text: The frequency of respiratory infections as an infection in cancer patients is high. Several factors increase the risk of respiratory infections in cancer patients. These include airway obstruction, disruption of mucosal surfaces and treatment-related factors, such as radiation and chemotherapy. It has also been reported that the frequency and mortality of drug-induced lung injury associated with antineoplastic drugs are high in Japan. In the diagnosis and treatment of drug-induced lung disease, it is important to distinguish pneumonia as an infectious disorder. Immune checkpoint inhibitors(ICIs)have been widely introduced to treat several types of malignancies. It is currently believed that ICIs do not increase the risk of infections. However, immune-related adverse events(irAEs)induced by ICIs may necessitate the administration of immunosuppressive agents, which could cause opportunistic infections. Furthermore, several reports describe reactivation tuberculosis without irAEs or immunosuppressants. Further studies are needed to prevent the development of tuberculosis in patients receiving ICIs.


title: One Bioregion/One Health: An Integrative Narrative for Transboundary Planning along the US-Mexico Border.

text: Global megatrends-including climate change, food and water insecurity, economic crisis, large-scale disasters and widespread increases in preventable diseases-are motivating a bioregionalisation of planning in city-regions around the world. Bioregionalisation is an emergent process. It is visible where societies have begun grappling with complex socio-ecological problems by establishing place-based (territorial) approaches to securing health and well-being. This article examines a bioregional effort to merge place-based health planning and ecological restoration along the US-Mexico border. The theoretical construct underpinning this effort is called One Bioregion/One Health (OBROH). OBROH frames health as a transborder phenomenon that involves human-animal-environment interactions. The OBROH approach aims to improve transborder knowledge networking, ecosystem resilience, community participation in science-society relations, leadership development and cross-disciplinary training. It is a theoretically informed narrative to guide action. OBROH is part of a paradigm shift evident worldwide; it is redefining human-ecological relationships in the quest for healthy place making. The article concludes on a forward-looking note about the promise of environmental epidemiology, telecoupling, ecological restoration, the engaged university and bioregional justice as concepts pertinent to reinventing place-based planning.


Pandemic Influenza: public health preparedness for the next global health emergency.


title: Drug-induced tardive motor syndromes.

text: Drug-induced tardive motor syndromes (TMS) is a group of disorders, characterized by involuntary movements of the tongue, face, lips, trunk and extremities, occurring after long-term exposure to a variety of pharmacological agents, mostly neuroleptics. The diagnosis of TMS requires exposure to dopamine receptor blocking agents for a period at least of 3 months, although for people over 60 years old the necessary exposure period is limited to 1 month. The exact pathophysiology still remains obscure. The aim of this article is to review the phenomenology, epidemiology and treatment options of the TMS, as clinically distinct movement disorders. TMS include tardive dyskinesia, which is the most common movement disorder, tardive dystonia, tardive akathisia, tardive Tourettism, tardive tremor and tardive myoclonus as well as some specific syndromes less often presented such as Pisa, Meige and Rabbit syndromes. Prevention remains the cornerstone in good clinical practice. Preventive approach requires thorough diagnostic process with frequent reviews in order to determine the necessity of use and dosing of neuroleptic treatment. Clinical vigilance for early detection of signs of TMS as well as recording of early extrapyramidal side-effects in the patient's history is needed, as these may predict the occurrence of TMS. In case of occurrence of TMS, gradual discontinuation of the offending agent is required. Therapeutic interventions include the administration of the following agents: atypical antipsychotics (mainly clozapine), benzodiazepines, vitamin E, reserpine, tetrabenazine, anticholinergics, botulinum toxin A. The early management of TMS is crucial for the patients' better clinical outcome and improved quality of life.


Revised nomenclature for coronavirus structural proteins, mRNAs and genes.


title: [Viruses in the feces of patients with viral hepatitis and other enterovirus infections].

text: Direct and immune electron microscopy was used to determine the frequency of finding of hepatitis A virus (HAV) and other viral agents in feces of patients in relation to the diagnosis and epidemiological situation. HAV-containing excretions from patients were analysed ultrastructurally. The highest frequency of HAV detection was established in patients in a water-borne and food-borne outbreak of hepatitis A (HA) and was 40.9% and 36.9%, respectively. In patients with HA diagnosis in the period of a seasonal rise of HA incidence HAV particles were found in 11.8%, and in the interseasonal period in 5%. Apart from HAV particles, in a small per cent of patients with HA diagnosis adenovirus and enterovirus particles were found. In patients with the disease diagnosed as hepatitis B (HB) only adenovirus and enterovirus particles were found. In contrast to the patients with HAV and HB diagnosis, the patients who were combined into a conditional "non-hepatitis" group were found to have, in addition to HAV (in the period of seasonal rise of HA incidence), adenovirus and enterovirus particles, also particles of astrovirus, coronavirus, and rotavirus. In fecal specimens of patients containing typical HAV particles structures were found resembling individual fragments of empty HAV particles, antibody-covered 17-22 nm and 27 nm spherical structures poorly reacting with antibody.


title: [Covid-19 - deaths and analysis].

text: Mortality from Covid-19 is monitored in detail and compared between countries with different strategies against the virus. There is, however, often a lack of understanding of what is required in terms of measures and interpretation to enable correct comparisons. The number of deaths from Covid-19 is affected by the testing strategy and many other things that differ between countries. Therefore, today, the most reliable source for monitoring and comparing mortality from Covid-19 is total mortality. In Sweden, there is good correspondence of Covid-19 deaths and total mortality, with a tendency to a higher total mortality indicating some under-reporting of Covid-19 mortality.


Beijing/W Mycobacterium tuberculosis in Italy.


title: Borna Disease Virus Assembles Porous Cage-like Viral Factories in the Nucleus.

text: Animal-derived RNA viruses frequently generate viral factories in infected cells. However, the details of how RNA viruses build such intracellular structures are poorly understood. In this study, we examined the structure and formation of the viral factories, called viral speckle of transcripts (vSPOTs), that are produced in the nuclei of host cells by Borna disease virus (BDV). Super-resolution microscopic analysis showed that BDV assembled vSPOTs as intranuclear cage-like structures with 59-180-nm pores. The viral nucleoprotein formed the exoskeletons of vSPOTs, whereas the other viral proteins appeared to be mainly localized within these structures. In addition, stochastic optical reconstruction microscopy revealed that filamentous structures resembling viral ribonucleoprotein complexes (RNPs) appeared to protrude from the outer surfaces of the vSPOTs. We also found that vSPOTs disintegrated into RNPs concurrently with the breakdown of the nuclear envelope during mitosis. These observations demonstrated that BDV generates viral replication factories whose shape and formation are regulated, suggesting the mechanism of the integrity of RNA virus persistent infection in the nucleus.


title: Real-time PCR for diagnosis of human bocavirus infections and phylogenetic analysis.

text: The human bocavirus (hBoV) was first described in 2005 in respiratory tract samples. The clinical relevance of hBoV is still unclear. The aim of our study was to establish a real-time PCR assay for the detection and quantification of hBoV DNA, to apply the real-time assay for the analysis of stool and serum samples for the presence of hBoV DNA, and to perform a phylogenetic analysis of the hBoV positive samples. A total of 834 nasopharyngeal aspirates (NPA), 10 serum samples, and 31 stool samples of children with acute respiratory diseases were retrospectively tested. For phylogenetic analysis, 968 bp of the VP2 gene were sequenced from 69 hBoV-positive NPA samples. The qualitative results of the real-time hBoV PCR were in good agreement with a conventional hBoV PCR. We found that 12% of the NPA were positive for hBoV DNA. The median viral load in the NPA was 4.9 x 10(3) copies/ml (range, 2.7 x 10 degrees to 1.5 x 10(11) copies/ml). There was no difference of the hBoV load in NPA between children with or without known coinfection, but the load was significantly higher in children with bronchitis than in children with the diagnosis of febrile seizures. hBoV DNA was found in 1 of 10 serum samples and in 14 of 31 stool samples. hBoV sequence identity was >99% in the VP2 region. In conclusion, hBoV DNA can be found in NPA samples at very high titers. In addition to being found in the respiratory tract, hBoV was found in stool samples. The clinical relevance of these findings remains to be determined.


title: Experimental sialodacryoadenitis virus infection in severe combined immunodeficient mice.

text: Mice with a severe combined immunodeficiency in B and T lymphocytes and natural killer cells (SCID-beige) were inoculated intranasally with sialodacryoadenitis (SDA) virus, a coronavirus of rats. Animals were killed at designated intervals and tissues were examined for evidence of viral infection by light microscopy and immunofluorescence microscopy. Based on these criteria, there was no evidence that these immunodeficient mice were susceptible to infection with SDA virus.


Delayed immunisation and risk of pertussis in infants: unmatched case-control study.


title: Association of diarrhea in cattle with torovirus infections on farms.

text: An epidemiologic survey was performed to determine the incidence of torovirus infections in 2 disease entities of cattle: diarrhea of replacement calves up to 2 months old, and winter dysentery of adult cattle. Samples were obtained from 187 diarrheal and 115 healthy calves from 15 farms, as well as 149 diarrheal and 67 healthy cows from 27 farms with or without winter dysentery. Enzyme-linked immunosorbent assays for detection of torovirus, rotavirus, and coronavirus antigen in feces, and of torovirus and coronavirus antibodies in serum were used to monitor infections in these groups. Torovirus was detected in 9 of the 15 farms in the study, and in 6% of calves with diarrhea, which was significantly higher than in healthy calves (2%). Seroconversion to torovirus was found significantly more often after winter dysentery episodes than on farms without a disease history; coronavirus seroconversion was less common.


title: Prostheses Used in Laparoscopic Inguinal Hernia Repair: Biocompatibility, Postoperative Complications and Quality of Life - Review of the Literature.

text: INTRODUCTION There is a continuous concern about meshes used in laparoscopic inguinal hernia repair, which mainly relates to their biocompatibility and ability to decrease postoperative complications and recurrence rate; in other words, efforts are made to find the "ideal" prosthesis. OBJECTIVES To evaluate different prostheses used in laparoscopic inguinal hernia repair in terms of biocompatibility, postoperative complications, recurrence rate and quality of life, so that all the features could lead us to the "ideal" mesh. MATERIAL AND METHOD Randomized controlled trials, reviews, prospective and retrospective studies, retrospective cross-sectional and experimental studies on animals published between 2000 and 2016 were analysed with respect to several features of a mesh: biocompatibility, postoperative complications, recurrence rate and quality of life. OUTCOMES The most common comparison is between heavy-weight and light-weight mesh used in laparoscopic inguinal hernia repair. Experimental studies try to discover the "ideal" prosthesis, which could provide improved biocompatibility, low postoperative complications, decreased recurrence rate and good quality of life. The most commonly used mesh that meats the characteristics of an "ideal" prosthesis is a light-weight monofilament macroporous polypropylene mesh, with a minimum tensile strength >16 N/cm², measuring 10x15 cm. CONCLUSIONS Published data show that the "ideal" prosthesis used in laparoscopic inguinal hernia repair has not been discovered yet. Regarding heavy- or light-weight meshes, there is no significant effect on recurrence, acute or chronic pain, incidence of seroma or return to daily activity and quality of life (1).


title: [Genome-wide scan reveals the molecular mechanisms of functional differentiation of Myotis lucifugus and Pteropus vampyrus].

text: The physiological and behavioral differences between Myotis lucifugus and Pteropus vampyrus should be attributed to molecular mechanisms and deserve intensive investigation. We conducted genome-wide scan for coding sequences from the orthologue genes of seven mammalian species. Selection analyses were carried out by setting the branches leading to Myotis lucifugus and Pteropus vampyrus as foreground branches, respectively. Enrichment analyses were conducted for positively selected genes. Our results indicated that more genes exhibited positive selection in Myotis lucifugus than that in Pteropus vampyrus. The positively selected genes of the two species were enriched in different functions. The differences between Myotis lucifugus and Pteropus vampyrus represented their differentiation in biological functions, especially the functions of immunity, motor ability, energetic metabolism and sensory organ development.


docid: mofuloaa

title: Nephrotic syndrome under treatment with dasatinib: be aware of a possible adverse drug reaction.

text: The protein kinase inhibitor dasatinib, targeting BCR-ABL and Src family kinases, is used in chronic myeloid leukaemia and Philadelphia-chromosome positive acute lymphoblastic leukaemia. The Netherlands Pharmacovigilance Centre Lareb has received one report of nephrotic syndrome associated with the use of dasatinib. With some other protein kinase inhibitors, targeting vascular endothelial growth factor, nephrotic syndrome is a well-known adverse drug reaction. The Dutch and European pharmacovigilance databases and scientific literature contain several cases indicating a causal relationship between dasatinib and nephrotic syndrome. Nephrotic syndrome was recently added to the list of adverse drug reactions in the Dutch summary of product characteristics for dasatinib. It is important to recognise the possibility of this adverse drug reaction when a patient develops nephrotic syndrome under treatment with dasatinib.


title: CD147 and its interacting proteins in cellular functions.

text: CD147 (basigin, EMMPRIN, neurothelin, M6, HAb18G, etc.), a transmembrane glycoprotein, has a broad expression pattern on various epithelial cells with some differences between species, e.g. rat, mouse, chicken and human, but is highly enriched on the surface of cancer cells of epithelial origin such as lung cancer, breast cancer and hepatoma cells. The CD147 antigen consists of two IgSF domains, a transmembrane sequence containing a charged residue (Glu) and a cytoplasmic domain of 40 residues. The particular structural features suggest that it is involved in protein-protein interactions. Although the interacting molecules are still not well known due to unavailability of the 3D structure of CD147, adhesion, coimmunoprecipitation and other studies recently suggest that several proteins, including integrins, cyclophilins, MCT, etc., interact with CD147 as its ligand or receptor candidates to mediate a wide range of cellular functions.


title: Cryptic properties of a cluster of dominant flavivirus cross-reactive antigenic sites.

text: A number of flaviviruses are important human pathogens, including yellow fever, dengue, West Nile, Japanese encephalitis, and tick-borne encephalitis (TBE) viruses. Infection with or immunization against any of these viruses induces a subset of antibodies that are broadly flavivirus cross-reactive but do not exhibit significant cross-neutralization. Nevertheless, these antibodies can efficiently bind to the major envelope protein (E), which is the main target of neutralizing and protective antibodies because of its receptor-binding and membrane fusion functions. The structural basis for this phenomenon is still unclear. In our studies with TBE virus, we have provided evidence that such cross-reactive antibodies are specific for a cluster of epitopes that are partially occluded in the cage-like assembly of E proteins at the surfaces of infectious virions and involve-but are not restricted to-amino acids of the highly conserved internal fusion peptide loop. Virus disintegration leads to increased accessibility of these epitopes, allowing the cross-reactive antibodies to bind with strongly increased avidity. The cryptic properties of these sites in the context of infectious virions can thus provide an explanation for the observed lack of efficient neutralizing activity of broadly cross-reactive antibodies, despite their specificity for a functionally important structural element in the E protein.


docid: gqisbvbn

title: Survey of infectious agents involved in acute respiratory disease in finishing pigs.

text: Outbreaks of respiratory disease constitute a major health problem in herds of finishing pigs and their aetiology often remains unclear. In this study, 16 outbreaks of respiratory disease with acute clinical signs in finishing pigs were investigated to determine which infectious agents were involved. From each herd four diseased and two clinically healthy pigs were examined pathologically and for the presence of viruses, bacteria and mycoplasmas. In addition, paired blood samples from 10 groupmates of the diseased pigs were tested for antibodies against commonly known causal agents of respiratory disease. A clear diagnosis was possible in 12 of the 16 outbreaks. Seven were due to an infection with influenza virus and five were due to an infection with Actinobacillus pleuropneumoniae. A combination of influenza virus and A pleuropneumoniae may have caused one other outbreak, but no clear cause could be established for the other three outbreaks.


docid: brrnrzew

title: Epidemiology and Clinical Features of Coronavirus Disease 2019 in Moroccan Children.

text: OBJECTIVES This study aims to analyze the epidemiological and clinical features of coronavirus disease 19 (COVID-19) in a Moroccan pediatric population. METHODS A retrospective study of a cohort of 74 children with RT-PCR confirmed COVID-19. We collected information on clinical and laboratory features of all children (age <18 years) admitted between 2 March, 2020 and 1 April, 2020. RESULTS The mean (SD) age of the 74 children (40 girls) was 7 (1.5) years. The mean (SD) time from illness onset to diagnosis was 2 (1) days. 54 children were asymptomatic, while eight had fever, and five cases had cough. Recovery was after a mean (SD) of 12 (1) days. CONCLUSION COVID-19 was mostly mild in the pediatric population in Morocco.


Public health and epidemiological research: a blind spot among the European Union priorities?


title: Respiratory syncytial virus polypeptides. IV. The oligosaccharides of the glycoproteins.

text: The cell-associated glycoproteins of respiratory syncytial (RS) virus included GP1 (90K), VP70 (70K), VGP48 (48K) and GP26 (26K). Although present in infected cells, there was no VP70 in purified virus. Trypsin treatment of infected cells removed 80 to 90% of VP70 as well as its products VGP48 and GP26. This suggested that most of the VP70 in the cell is located on the plasma membrane. The glycoproteins of purified RS virus (GP1, VGP48 and GP26) contain mannose, galactose and fucose as well as glucosamine, but the quantity of mannose in GP1 is low when compared to that of the other three sugars. The effects that follow the treatment of infected cells with the glycosylation inhibitors tunicamycin and monensin, and the treatment of the immunoprecipitated product of pulse-chase experiments with endonuclease H demonstrated that VP70 and its products contained N-linked oligosaccharides, and that the oligosaccharides of the mature VGP48 subunit were of the complex type, while GP1 contained both N- and O-linked oligosaccharides. The non-glycosylated forms of VP70 and GP1 have estimated mol. wt. of 50K and 33K respectively. Therefore, the carbohydrate contribution to the mol. wt. of VP70 and GP1, as determined by PAGE, was equivalent to 20K for the former and 57K for the latter. The majority of the GP1 oligosaccharides were O-linked, a form of sugar linkage not previously found among paramyxoviruses.


title: Endoluminal abdominal aortic aneurysm repair: the latest advances in prevention of distal endograft migration and type 1 endoleak.

text: Endovascular abdominal aortic aneurysm repair (EVAR) is an attractive alternative to open surgical repair. Distal endograft migration and type 1 endoleak are recognized to be the 2 main complications of EVAR. First-generation endografts had a stronger propensity for distal migration, modular component separation, thrombosis, and loss of structural integrity. Substantial progress has been made in recent years with 2nd- and 3rd-generation devices to prevent these complications. Some of the most common predictors of endograft failure are angulated and short infrarenal necks, large-diameter necks, and thrombus in the aneurysmal sac. The purpose of this study is to describe and review our experience in using innovative techniques and a newer generation of endografts to prevent distal migration and type 1 endoleak in patients with challenging infrarenal neck anatomy. The use of these innovative EVAR techniques and the new generation of endografts in patients with challenging infrarenal neck anatomy has yielded encouraging procedural and intermediate-term results.


title: Neuromodulation device therapy for treatment of hypertensive heart disease.

text: Hypertensive heart disease (HHD) is the leading cause of mortality and morbidity in the United States. Despite the availability of medical therapy it remains a challenge to treat. Autonomic nervous system imbalance resulting in overactivity of the sympathetic nervous system and under activity of the parasympathetic nervous system is integral in the development of hypertension and ultimately the development of HHD. Emerging data suggest that neuromodulation device therapy for treatment of HHD is promising.


docid: gxarfjkn

title: Prevalence of IgG antibody to SARS-associated coronavirus in animal traders--Guangdong Province, China, 2003.

text: Severe acute respiratory syndrome (SARS) was identified in 2003 as an infectious disease caused by the SARS-associated coronavirus (SARS-CoV), a member of the coronavirus family not observed previously in humans. Because its sequence data differ from that of known human coronaviruses, SARS-CoV is suspected to have crossed the species barrier between an animal host and humans. The SARS outbreak began in China's Guangdong Province, where approximately 1,500 probable cases were identified during November 2002-June 2003. Detection of SARS-like coronavirus has been reported previously in masked palm civets (sometimes called civet cats) and a raccoon dog for sale in a live animal market in Shenzhen municipality. This report summarizes results of an investigation conducted by public health authorities in Guangdong Province, which compared the seroprevalence of SARS-CoV IgG antibody in animal traders (i.e., workers in live animal markets) with that of persons in control groups. The results indicated that 13% of the animal traders, none of whom had SARS diagnosed, had IgG antibody to SARS-CoV, compared with 1%-3% of persons in three control groups. Although the results provide indirect support for the hypothesis of an animal origin for SARS, they also underscore the need for detailed patient histories and more focused animal studies to confirm an animal origin for SARS.


Evaluating Routine Upper Gastrointestinal Series after Laparoscopic Sleeve Gastrectomy.


title: Design and Coordination Kinematics of an Insertable Robotic Effectors Platform for Single-Port Access Surgery.

text: Single port access surgery (SPAS) presents surgeons with added challenges that require new surgical tools and surgical assistance systems with unique capabilities. To address these challenges, we designed and constructed a new insertable robotic end-effectors platform (IREP) for SPAS. The IREP can be inserted through a Ø15 mm trocar into the abdomen and it uses 21 actuated joints for controlling two dexterous arms and a stereo-vision module. Each dexterous arm has a hybrid mechanical architecture comprised of a two-segment continuum robot, a parallelogram mechanism for improved dual-arm triangulation, and a distal wrist for improved dexterity during suturing. The IREP is unique because of the combination of continuum arms with active and passive segments with rigid parallel kinematics mechanisms. This paper presents the clinical motivation, design considerations, kinematics, statics, and mechanical design of the IREP. The kinematics of coordination between the parallelogram mechanisms and the continuum arms is presented using the pseudo-rigid-body model of the beam representing the passive segment of each snake arm. Kinematic and static simulations and preliminary experiment results are presented in support of our design choices.


title: Expression of ICAM-1 in nasal epithelium and levels of soluble ICAM-1 in nasal lavage fluid during human experimental rhinovirus infection.

text: Most rhinovirus serotypes use intercellular adhesion molecule-1 (ICAM-1) as the receptor to enter cells, but ICAM-1 expression has not been detected on normal nasal epithelial cells. During experimental rhinovirus infection, expression of ICAM-1 on nasal epithelial cells was examined with immunohistochemical staining of nasal scrape biopsy specimens, and levels of soluble ICAM-1 in nasal lavage fluid were measured by sandwich enzyme-linked immunosorbent assay technique. Expression of ICAM-1 on nasal epithelial cells increased following inoculation in 20 of 23 infected subjects. The median number of ICAM-1-positive cells per 6.25-mm(2) area of stained biopsy specimen was 0 in control samples (day 20 or 33 after inoculation), and in those without infection, 6 on day 1 (P< or =.05), 14.5 on day 3 (P< or =.01), 1.5 on day 5, and 0 on day 9. In a different group of volunteers, soluble ICAM-1 in nasal lavage fluid was higher on days 1 and 3 compared with preinoculation levels (P< or =.001), but only 11 of 23 infected subjects had a 2-fold or greater increase. Up-regulation of ICAM-1 receptor expression on nasal epithelial cells occurred within 24 hours after inoculation in experimental rhinovirus infections (prior to onset of symptoms) and declined promptly by day 5.


COVID-19: early reflections.


title: Toxicological safety evaluation of DNA plasmid vaccines against HIV-1, Ebola, Severe Acute Respiratory Syndrome, or West Nile virus is similar despite differing plasmid backbones or gene-inserts.

text: The Vaccine Research Center has developed a number of vaccine candidates for different diseases/infectious agents (HIV-1, Severe Acute Respiratory Syndrome virus, West Nile virus, and Ebola virus, plus a plasmid cytokine adjuvant-IL-2/Ig) based on a DNA plasmid vaccine platform. To support the clinical development of each of these vaccine candidates, preclinical studies were performed to screen for potential toxicities (intrinsic and immunotoxicities). All treatment-related toxicities identified in these repeated-dose toxicology studies have been confined primarily to the sites of injection and seem to be the result of both the delivery method (as they are seen in both control and treated animals) and the intended immune response to the vaccine (as they occur with greater frequency and severity in treated animals). Reactogenicity at the site of injection is generally seen to be reversible as the frequency and severity diminished between doses and between the immediate and recovery termination time points. This observation also correlated with the biodistribution data reported in the companion article (Sheets et al., 2006), in which DNA plasmid vaccine was shown to remain at the site of injection, rather than biodistributing widely, and to clear over time. The results of these safety studies have been submitted to the Food and Drug Administration to support the safety of initiating clinical studies with these and related DNA plasmid vaccines. Thus far, standard repeated-dose toxicology studies have not identified any target organs for toxicity (other than the injection site) for our DNA plasmid vaccines at doses up to 8 mg per immunization, regardless of disease indication (i.e., expressed gene-insert) and despite differences (strengths) in the promoters used to drive this expression. As clinical data accumulate with these products, it will be possible to retrospectively compare the safety profiles of the products in the clinic to the results of the repeated-dose toxicology studies, in order to determine the utility of such toxicology studies for signaling potential immunotoxicities or intrinsic toxicities from DNA vaccines. These data build on the biodistribution studies performed (see companion article, Sheets et al., 2006) to demonstrate the safety and suitability for investigational human use of DNA plasmid vaccine candidates for a variety of infectious disease prevention indications.


title: Respiratory viruses and influenza-like illness: a survey in the area of Rome, winter 2004-2005.

text: Limited information is available on the viral aetiology of influenza-like illness (ILI) in Southern European countries. Hereby we report the main findings of a survey conducted in the area of Rome during the 2004-2005 winter season.ILI cases were defined as individuals with fever >37.5 degrees C and at least one constitutional symptom and one respiratory symptom, recruited during the survey period. Influenza and other respiratory viruses were identified using polymerase chain reaction (PCR) on throat swabs. Basic individual information was collected through a standard form. Of 173 ILI cases enrolled, 74 tested positive for one virus, and two tested positive for two viruses. Overall, 33.5% of the cases were positive for influenza viruses, 5.2% for adenoviruses, 3.5% for parainfluenza viruses, 1.7% for coronaviruses, and 1.2% for the respiratory syncitial virus. The proportion of influenza virus detection was higher in the 'high influenza activity' period. The distribution of viral agents varied across age groups, influenza viruses being more likely to be detected in younger patients. Viral pathogens were identified in less than 50% of ILI cases occurred during a high activity influenza season. The detection of other than influenza viruses was sporadic, without evidence of large outbreaks due to specific agents.


Why I won't see you on the barricades: Disability and COVID-19.


title: Detection of feline coronavirus infection in captive cheetahs (Acinonyx jubatus) by polymerase chain reaction.

text: Feline coronavirus genetic elements were detected by polymerase chain reaction from blood, fecal samples, and effusive fluid collected from 33 cheetahs in the U.S.A. Feline coronavirus-specific serum antibodies were also measured by indirect immunofluorescence. Ten cheetahs were positive for viral shedding by polymerase chain reaction, whereas 13 were seropositive by immunofluorescence. Results of serology did not consistently correlate with shedding of virus, and the capture antigen used for detection of feline coronavirus-specific antibodies had a significant impact on results. Testing of samples from one population over a 1-yr period indicated chronic infection in some animals. These relatively healthy carrier animals were a source of virus for contact animals. Screening programs in cheetah populations for feline coronavirus infection may be most reliable if a combination of serologic analysis and viral detection by polymerase chain reaction is used.


title: Amino acids from both N-terminal hydrophobic regions of the Lassa virus envelope glycoprotein GP-2 are critical for pH-dependent membrane fusion and infectivity.

text: Lassa virus glycoprotein 2 (LASV GP-2) belongs to the class I fusion protein family. Its N terminus contains two stretches of highly conserved hydrophobic amino acids (residues 260-266 and 276-298) that have been proposed as N-terminal or internal fusion peptide segments (N-FPS, I-FPS) by analogy with similar sequences of other viral glycoproteins or based on experimental data obtained with synthetic peptides, respectively. By using a pH-dependent, recombinant LASV glycoprotein mediated cell-cell fusion assay and a retroviral pseudotype infectivity assay, an alanine scan of all hydrophobic amino acids within both proposed FPSs was performed. Fusogenicity and infectivity were correlated, both requiring correct processing of the glycoprotein precursor. Most point mutations in either FPS accounted for reduced or abolished fusion or infection, respectively. Some mutations also had an effect on pre-fusion steps of virus entry, possibly by inducing structural changes in the glycoprotein. The data demonstrate that several amino acids from both hydrophobic regions of the N terminus, some of which (W264, G277, Y278 and L280) are 100 % conserved in all arenaviruses, are involved in fusogenicity and infectivity of LASV GP-2.


title: Offering lung cancer screening to high-risk medicare beneficiaries saves lives and is cost-effective: an actuarial analysis.

text: BACKGROUND By a wide margin, lung cancer is the most significant cause of cancer death in the United States and worldwide. The incidence of lung cancer increases with age, and Medicare beneficiaries are often at increased risk. Because of its demonstrated effectiveness in reducing mortality, lung cancer screening with low-dose computed tomography (LDCT) imaging will be covered without cost-sharing starting January 1, 2015, by nongrandfathered commercial plans. Medicare is considering coverage for lung cancer screening. OBJECTIVE To estimate the cost and cost-effectiveness (ie, cost per life-year saved) of LDCT lung cancer screening of the Medicare population at high risk for lung cancer. METHODS Medicare costs, enrollment, and demographics were used for this study; they were derived from the 2012 Centers for Medicare & Medicaid Services (CMS) beneficiary files and were forecast to 2014 based on CMS and US Census Bureau projections. Standard life and health actuarial techniques were used to calculate the cost and cost-effectiveness of lung cancer screening. The cost, incidence rates, mortality rates, and other parameters chosen by the authors were taken from actual Medicare data, and the modeled screenings are consistent with Medicare processes and procedures. RESULTS Approximately 4.9 million high-risk Medicare beneficiaries would meet criteria for lung cancer screening in 2014. Without screening, Medicare patients newly diagnosed with lung cancer have an average life expectancy of approximately 3 years. Based on our analysis, the average annual cost of LDCT lung cancer screening in Medicare is estimated to be $241 per person screened. LDCT screening for lung cancer in Medicare beneficiaries aged 55 to 80 years with a history of ≥30 pack-years of smoking and who had smoked within 15 years is low cost, at approximately $1 per member per month. This assumes that 50% of these patients were screened. Such screening is also highly cost-effective, at <$19,000 per life-year saved. CONCLUSION If all eligible Medicare beneficiaries had been screened and treated consistently from age 55 years, approximately 358,134 additional individuals with current or past lung cancer would be alive in 2014. LDCT screening is a low-cost and cost-effective strategy that fits well within the standard Medicare benefit, including its claims payment and quality monitoring.


title: Debate on MERS-CoV respiratory precautions: surgical mask or N95 respirators?

text: Since the emergence of Middle East respiratory syndrome coronavirus (MERS-CoV) in mid-2012, there has been controversy over the respiratory precaution recommendations in different guidelines from various international bodies. Our understanding of MERS-CoV is still evolving. Current recommendations on infection control practices are heavily influenced by the lessons learnt from severe acute respiratory syndrome. A debate on respiratory precautions for MERS-CoV was organised by Infection Control Association (Singapore) and the Society of Infectious Disease (Singapore). We herein discuss and present the evidence for surgical masks for the protection of healthcare workers from MERS-CoV.


title: Perinatal changes of cardiac troponin-I in normal and intrauterine growth-restricted pregnancies.

text: Intrauterine growth restriction (IUGR) implies fetal hypoxia, resulting in blood flow redistribution and sparing of vital organs (brain, heart). Serum cardiac Troponin-I (cTnI), a well-established marker of myocardial ischaemia, was measured in 40 mothers prior to delivery, the doubly clamped umbilical cords (representing fetal state), and their 20 IUGR and 20 appropriate-for-gestational-age (AGA) neonates on day 1 and 4 postpartum. At all time points, no differences in cTnI levels were observed between the AGA and IUGR groups. Strong positive correlations were documented between maternal and fetal/neonatal values (r > or = .498, P < or = .025 in all cases in the AGA and r > or = .615, P < or = .009 in all cases in the IUGR group). These results may indicate (a) normal heart function, due to heart sparing, in the IUGR group (b) potential crossing of the placental barrier by cTnI in both groups.


title: Pathological and immunological effects of ingesting L-tryptophan and 1,1'-ethylidenebis (L-tryptophan) in Lewis rats.

text: The eosinophilia-myalgia syndrome (EMS) has been associated with ingestion of L-tryptophan (L-TRP) produced by a single manufacturer. Epidemiological data implicated 1,1'-ethylidenebis (L-tryptophan) (EBT) (peak 97 or peak E) as a possible etiologic agent. We showed previously that Lewis rats treated with the L-TRP implicated in EMS develop fasciitis and perimyositis similar to those seen in human EMS. We now report the pathology associated with the treatment of Lewis rats with synthetic EBT and/or L-TRP. All animals treated for 6 wk with case-associated L-TRP or EBT developed significant myofascial thickening, compared with animals in the vehicle control and control L-TRP groups. However, even those animals receiving the control L-TRP showed a mild but significant increase in the thickness of the myofascia, compared with vehicle-treated control animals. All animals except vehicle controls also exhibited significant pancreatic pathology, including fibrosis and acinar changes. Only animals treated with case-associated L-TRP for 6 wk showed evidence of immune activation with increased frequency of CD8, Ia, and IL-2 receptor-positive cells in the peripheral blood. Animals receiving L-TRP or EBT for < 6 wk did not show significant differences in myofascial thickness, although these animals did show pancreatic acinar changes. Although these results demonstrate for the first time the pathological effects of EBT, they do not rule out the possibility that other impurities in the EMS-case-associated L-TRP may also contribute to some of the features of EMS.


title: An outbreak of severe acute respiratory syndrome among hospital workers in a community hospital in Hong Kong.

text: BACKGROUND During outbreaks, hospital workers are at high risk for nosocomial infection with severe acute respiratory syndrome (SARS)-associated coronavirus. OBJECTIVE To examine how hospital workers became infected and whether they transmit the virus to their families. DESIGN Retrospective descriptive study. SETTING 529-bed community hospital in Hong Kong. PATIENTS 40 hospital workers infected with SARS-associated coronavirus over a 6-week period (25 March through 5 May 2003). MEASUREMENTS Percentage of infected hospital workers according to job category. RESULTS The cumulative incidence was highest among health care assistants, followed by physicians and nurses (8%, 5%, and 4%, respectively). Most hospital workers were infected from direct contact with patients with SARS, who primarily were in general wards and had unsuspected infection. At the time of contact, all hospital workers had used masks but not necessarily other protective devices. Affected hospital workers did not infect their families. CONCLUSION Before isolation of all patients with clinically confirmed or suspected SARS, routine use of several protective devices, and training of staff in infection control, many health care workers were infected with SARS from patients with unsuspected cases.


title: Induction of glial cell MHC antigen expression in neurotropic coronavirus infections. Characterization of the H-2-inducing soluble factor elaborated by infected brain cells.

text: Neurotropic coronavirus (mouse hepatitis virus strain A59) infection induces major histocompatibility complex class I (H-2) surface antigens on oligodendrocytes and astrocytes, cells that do not normally express detectable MHC antigens on their surface. The induction on MHC antigen expression potentially allows immunocytes to interact with infected glial cells and may play a critical role in the development of virus-induced, immune-mediated demyelination in the central nervous system, a possible model of human multiple sclerosis. In this study, we characterized the soluble factor involved in MHC antigen induction, quantitated induction of MHC antigens, and analyzed the central nervous system cell type involved in the production of the factor. The H-2-inducing factor, most likely produced by astrocytes, was found to be nondialyzable, heat- and trypsin-sensitive, but resistant to treatment at pH 2.0. The m.w. of the factor was estimated as 50 to 100 kDa. Studies on fractionation by ultrafiltration and sucrose density gradient along with antibody-blocking experiments indicate that the factor is not interferon or virus particles.


Possible prevention of tick-borne relapsing fever in patients infected with Borrelia recurrentis.


title: Using Electromagnetic Navigation Bronchoscopy and Dye Injection to Aid in Video-Assisted Lung Resection.

text: Small (2 cm) peripheral lung lesions and ground glass opacities remain a difficult subset of lung lesions for the diagnosis and management of lung cancer. Surgical biopsy is more difficult for these lesions because intraoperative localization has to be made without the aid of direct visualization or manual palpation. Electromagnetic navigation bronchoscopy can be used in the operating room to identify a small peripheral lesion and marked using an injection of methylene blue, which can be seen on the visceral pleura of the lung. We present our initial experience using this technique. The sample was eight patients who had peripheral lesions with an average size of 19 mm. Surgical wedge biopsy was diagnostic in all cases, with an average procedure time of 28 minutes. There were no complications from this procedure. In conclusion, these data suggest that electromagnetic navigation bronchoscopy can be performed safely with high diagnostic accuracy by the operating thoracic surgeon, but further data are needed to establish its utility and safety.


[Digestive symptoms in coronavirus disease 2019 (COVID-19)].


title: Design and synthesis of dipeptidyl glutaminyl fluoromethyl ketones as potent severe acute respiratory syndrome coronovirus (SARS-CoV) inhibitors.

text: This paper describes the design and synthesis of dipeptidyl N,N-dimethyl glutaminyl fluoromethyl ketones (fmk) as severe acute respiratory syndrome coronovirus (SARS-CoV) inhibitors. The compounds were tested against SARS-CoV-induced cell death in Vero or CaCo2 cells as a measurement of the inhibiting effects of the compounds on the replication of the virus. Z-Leu-Gln(NMe(2))-fmk (6a) was found to be a potent inhibitor with low toxicity in cells, protecting cells with an EC(50) value of 2.5 microM and exhibiting a selectivity index of >40.


title: Microtiter plate based chemistry and in situ screening: a useful approach for rapid inhibitor discovery.

text: The use of libraries extracted from nature or constructed by combinatorial chemistry, have been widely appreciated in the drug discovery area. In this perspective, we present our contribution to the field of enzyme inhibitor discovery using a useful approach that allows diversification of a common core in a microtiter plate followed by in situ screening. Our method relies on an organic reaction that is highly selective, high yielding, amenable to the microscale and preferably can be performed in water. The core can be a designed molecule based on the structural and mechanistic information of the target, a compound with a weak binding affinity, or a natural product. Several reactions were found useful for this approach and were applied to the rapid discovery of potent inhibitors of representative enzymes.


title: Using models to identify routes of nosocomial infection: a large hospital outbreak of SARS in Hong Kong.

text: Two factors dominated the epidemiology of severe acute respiratory syndrome (SARS) during the 2002-2003 global outbreak, namely super-spreading events (SSE) and hospital infections. Although both factors were important during the first and the largest hospital outbreak in Hong Kong, the relative importance of different routes of infection has not yet been quantified. We estimated the parameters of a novel mathematical model of hospital infection using SARS episode data. These estimates described levels of transmission between the index super-spreader, staff and patients, and were used to compare three plausible hypotheses. The broadest of the supported hypotheses ascribes the initial surge in cases to a single super-spreading individual and suggests that the per capita risk of infection to patients increased approximately one month after the start of the outbreak. Our estimate for the number of cases caused by the SSE is substantially lower than the previously reported values, which were mostly based on self-reported exposure information. This discrepancy suggests that the early identification of the index case as a super-spreader might have led to biased contact tracing, resulting in too few cases being attributed to staff-to-staff transmission. We propose that in future outbreaks of SARS or other directly transmissible respiratory pathogens, simple mathematical models could be used to validate preliminary conclusions concerning the relative importance of different routes of transmission with important implications for infection control.


title: Transvaginal cholecystectomy versus single-incision laparoscopic cholecystectomy versus four-port laparoscopic cholecystectomy: a prospective cohort study.

text: OBJECTIVE This report describes the first prospective cohort study comparing transvaginal cholecystectomies (TVC) with single incision laparoscopic cholecystectomies (SILC) and four-port laparoscopic cholecystectomies (4PLC). METHODS Between May 2009 and August 2010, 14 patients underwent a TVC. These patients were compared with patients who underwent SILC (22 patients) or 4PLC (11 patients) in a concurrent, randomized, controlled trial. Demographic data, operative time, numerical pain scales, complications, and return to work were recorded. RESULTS Mean age (TVC: 33.5 ± 3.0 year; SILC: 38.4 ± 3.3 year; 4PLC: 35.5 ± 4.1 year; p = 0.58) and mean BMI (TVC: 28.8 ± 1.5 kg/m(2); SILC: 31.8 ± 1 kg/m(2); 4PLC: 31.4 ± 2.2 kg/m(2); p = 0.35) were not statistically significant. However, mean operative time (TVC: 67 ± 3.9 min; SILC: 48.9 ± 2.6 min; 4PLC: 42.3 ± 3.9 min; p < 0.001) was significantly longer for TVC. Numerical pain scales showed significantly lower pain scores on POD 1 and 3 for TVC compared with SILC and 4PLC (TVC: 4.1 ± 0.5 and 2.9 ± 0.7; SILC: 6.1 ± 0.5 and 5.3 ± 0.5; 4PLC: 5.7 ± 0.4 and 4.7 ± 0.3; p = 0.02) with equilibration of pain scores by days 14 and 30. Return to work (TVC: 6.4 ± 1.5 days; SILC: 13.1 ± 1.3 days; 4PLC: 14.1 ± 1.4 days; p < 0.001) also was significantly faster for patients in the TVC group. One conversion in the TVC group to a 4PLC was necessary due to adhesions within the pelvis. One dislodged IUD was seen and immediately replaced in the TVC group. One hernia was observed in the SILC group. CONCLUSIONS Transvaginal cholecystectomy is a safe and well-tolerated procedure with statistically significantly less pain at 1 and 3 days after surgery, with a faster return to work but longer operative times compared with single incision and four-port laparoscopic cholecystectomy.


title: Genetic diversity of avian infectious bronchitis viruses in Japan based on analysis of s2 glycoprotein gene.

text: To understand the genetic diversity of the S2 gene of infectious bronchitis viruses (IBV) isolated in Japan, we determined the nucleotide sequences of these IBVs using the reverse transcriptase polymerase chain reaction method coupled with direct sequencing. IBV isolated in Japan were classified into six different groups by phylogenetic analysis based on the S2 gene. However, the classification based on the S2 gene of IBV isolated in Japan was different for some of the strains from those obtained with our previous analysis of the S1 gene. This suggested that genetic recombination between the virus strains classified into different genetic groups had occurred in poultry, and that recombinant viruses might be epidemic in Japan.


title: Detection of airborne rhinovirus and its relation to outdoor air supply in office environments.

text: Rhinoviruses are major causes of morbidity in patients with respiratory diseases; however, their modes of transmission are controversial. We investigated detection of airborne rhinovirus in office environments by polymerase chain reaction technology and related detection to outdoor air supply rates. We sampled air from 9 A.M. to 5 P.M. each workday, with each sample run for 1 work week. We directly extracted RNA from the filters for nested reverse transcriptase-polymerase chain reaction analysis of rhinovirus. Nasal lavage samples from building occupants with upper respiratory infections were also collected. Indoor carbon dioxide (CO2 concentrations were recorded every 10 minutes as a surrogate for outdoor air supply. To increase the range of CO2 concentrations, we adjusted the outdoor air supply rates every 3 months. Generalized additive models demonstrated an association between the probability of detecting airborne rhinovirus and a weekly average CO2 concentration greater than approximately 100 ppm, after controlling for covariates. In addition, one rhinovirus from a nasal lavage contained an identical nucleic acid sequence similar to that in the building air collected during the same week. These results suggest that occupants in buildings with low outdoor air supply may have an increased risk of exposure to infectious droplet nuclei emanating from a fellow building occupant.


title: Technical Improvement Using a Three-Dimensional Video System for Laparoscopic Partial Nephrectomy.

text: BACKGROUND Laparoscopic partial nephrectomy is one of the major surgical techniques for small renal masses. However, it is difficult to manage cutting and suturing procedures within acceptable time periods. To overcome this difficulty, we applied a three-dimensional (3D) video system with laparoscopic partial nephrectomy, and evaluated its utility. MATERIALS AND METHODS We retrospectively enrolled 31 patients who underwent laparoscopic partial nephrectomy between November 2009 and June 2014. A conventional two-dimensional (2D) video system was used in 20 patients, and a 3D video system in 11. Patient characteristics and video system type (2D or 3D) were recorded, and correlations with perioperative outcomes were analyzed. RESULTS Mean age of the patients was 55.8±12.4, mean body mass index was 25.7±3.9 kg/m2, mean tumor size was 2.0±0.8 cm, mean R.E.N.A.L nephrometry score was 6.9±1.9, and clinical stage was T1a in all patients. There were no significant differences in operative time (p=0.348), pneumoperitoneum time (p=0.322), cutting time (p=0.493), estimated blood loss (p=0.335), and Clavien grade of >II complication rate (p=0.719) between the two groups. However, warm ischemic time was significantly shorter in the 3D group than the 2D group (16.1 min vs. 21.2min, p=0.021), which resulted from short suturing time (9.1 min vs. 15.2 min, p=0.008). No open conversion occurred in either group. CONCLUSIONS A 3D video system allows the shortening of warm ischemic time in laparoscopic partial nephrectomy and thus may be useful in improving the procedure.


title: Fringed membranous particles and viruses in faeces from healthy turkey poults and from poults with putative poult enteritis complex/spiking mortality.

text: The purpose of the present study was to use clinical epidemiologic tools to define the relationship between production performance data, virus particles, and intestinal fringed membranous particles (FMPs) in healthy turkey poults and in poults that were experiencing an outbreak of poult enteritis complex (PEC) and spiking mortality syndrome (SMS). Small and large intestines from flocks of healthy poults and poults with PEC/SMS were collected, processed, and examined for viruses. Production performance parameters were collected and analyzed. Hocks of turkeys with PEC/SMS in the present study had low survival expectancy (livability) and poor growth compared to their healthy turkey counterparts. The only significant association between sickness and intestinal virus was the presence of coronavirus.


title: Serological and molecular characterization of three enteric isolates of infectious bronchitis virus of chickens.

text: Three coronaviruses isolated from the intestines of laying chickens were partially characterized. Serological and molecular assays indicated that the enteric coronaviruses are infectious bronchitis virus (IBV) isolates. Although the three isolates were recovered from three unrelated chicken flocks, their RNase T1-resistant oligonucleotide fingerprints were almost identical. The three isolates were not neutralized by antisera specific to IBV serotype Connecticut, but their RNase T1-resistant oligonucleotide fingerprints closely matched the fingerprints of strain Conn-46, a Connecticut serotype. This and the co-fingerprinting data suggested that the three field isolates may have emerged from the Connecticut virus through mutation(s). The mutation(s) apparently involved the S1 protein gene that determines the virus serotype.


title: Transcription regulatory sequences and mRNA expression levels in the coronavirus transmissible gastroenteritis virus.

text: The transcription regulatory sequences (TRSs) of the coronavirus transmissible gastroenteritis virus (TGEV) have been characterized by using a helper virus-dependent expression system based on coronavirus-derived minigenomes to study the synthesis of subgenomic mRNAs. The TRSs are located at the 5' end of TGEV genes and include a highly conserved core sequence (CS), 5'-CUAAAC-3', that is essential for mediating a 100- to 1,000-fold increase in mRNA synthesis when it is located in the appropriate context. The relevant sequences contributing to TRS activity have been studied by extending the CS 5' upstream and 3' downstream. Sequences from virus genes flanking the CS influenced transcription levels from moderate (10- to 20-fold variation) to complete mRNA synthesis silencing, as shown for a canonical CS at nucleotide (nt) 120 from the initiation codon of the S gene that did not lead to the production of the corresponding mRNA. An optimized TRS has been designed comprising 88 nt from the N gene TRS, the CS, and 3 nt 3' to the M gene CS. Further extension of the 5'-flanking nucleotides (i.e., by 176 nt) decreased subgenomic RNA levels. The expression of a reporter gene (beta-glucuronidase) by using the selected TRS led to the production of 2 to 8 microg of protein per 10(6) cells. The presence of an appropriate Kozak context led to a higher level of protein expression. Virus protein levels were shown to be dependent on transcription and translation regulation.


Role of endoscopy in the management of GERD.


title: Incidence and characteristics of viral community-acquired pneumonia in adults.

text: BACKGROUND In adults, viral causes of community-acquired pneumonia (CAP) are poorly characterised. The aims of this study were to characterise the viral aetiology of CAP in adults by using an extensive array of viral diagnostic tests and to compare the characteristics of viral pneumonia with those of pneumococcal pneumonia. METHODS Adults admitted to Christchurch Hospital over a 1-year period with CAP were included in the study. Microbiological testing methods included blood and sputum cultures, urinary antigen testing for Streptococcus pneumoniae and Legionella pneumophila, antibody detection in paired sera and detection of respiratory viruses in nasopharyngeal swabs by immunofluorescence, culture and PCR. RESULTS Of 304 patients with CAP, a viral diagnosis was made in 88 (29%), with rhinoviruses and influenza A being the most common. Two or more pathogens were detected in 49 (16%) patients, 45 of whom had mixed viral and bacterial infections. There were no reliable clinical predictors of viral pneumonia, although several variables were independently associated with some aetiologies. The presence of myalgia was associated with pneumonia caused by any respiratory virus (OR 3.62, 95% CI 1.29 to 10.12) and influenza pneumonia (OR 190.72, 95% CI 3.68 to 9891.91). Mixed rhinovirus/pneumococcal infection was associated with severe disease. CONCLUSIONS Virus-associated CAP is common in adults. Polymicrobial infections involving bacterial and viral pathogens are frequent and may be associated with severe pneumonia.


title: Topology of double-membraned vesicles and the opportunity for non-lytic release of cytoplasm.

text: Infection of mammalian cells with several positive-strand RNA viruses induces double-membraned vesicles whose cytosolic surfaces serve as platforms for viral RNA replication. Our recent publication (Jackson et al. PLoS Biol 2005; 3:861-71) chronicled several similarities between poliovirus-induced membranes and autophagosomes, including induced co-localization of GFP-LC3 and LAMP1. Occasionally, the cytosolic lumen of these structures also contains viral particles; this likely results from wrapping of cytosol, which can contain high viral concentrations late in infection, by newly formed double membranes. Interestingly, RNAi treatment to reduce LC3 or Atg12p concentrations reduced yields of extracellular virus even more than intracellular virus. It is often assumed that exit of non-enveloped viruses such as poliovirus requires cell lysis. However, we hypothesize that autophagosome-like double-membranes, which can become single-membraned upon maturation, provide a long-sought mechanism for the observed non-lytic release of cytoplasmic viruses and possibly other cytoplasmic material resistant to the environment of maturing autophagosomes.


title: Human coronavirus OC43 interacts with major histocompatibility complex class I molecules at the cell surface to establish infection.

text: Human coronaviruses have been associated with common colds, diarrhea and enterocolitis, and have been implicated in multiple sclerosis. HLA class I molecules may play a critical role as receptor for OC43 because monoclonal antibody (mAb)W6/32 to HLA-A, -B and -C specificities completely blocks infectivity in human rhabdomyosarcoma (RD) cells. The role of HLA class 1 antigen as the virus receptor was examined using HLA-A3.1 stably transfected human plasma cells and untransfected HMY.C1R cells which do not express HLA-A and -B molecules. When the cells (5 x 10(6)) were infected at a multiplicity of one, the HLA.A3 transfected cells produced 10(8) PFU of virus whereas no replication occurred in the HMY.C1R cells mAb W6/32 reduced the virus yield by 99.9%. Cell membranes from HMY.C1R, HMY.A3 cells and chicken erythrocytes were biotinylated as live cells. Immunoprecipitation with polyclonal antiviral antibody to detect binding of biotinylated cell membranes to virus revealed that biotinylated HMY.A3 membranes coprecipitated with virus-antibody complexes when the immunoprecipitates were electrophoresed on SDS-PAGE gel, electroblotted and stained with Avidin-horseradish peroxidase. The results provide direct evidence that OC43 virus can recognize HLA class I as receptor on the cell surface.


title: Allergen challenge-induced acute exudation of IL-8, ECP and alpha2-macroglobulin in human rhinovirus-induced common colds.

text: Rhinovirus infections cause exacerbations of eosinophilic airway disease. The acute effects of allergen-challenge on nasal interleukin-8 (IL-8), eosinophil cationic protein (ECP), and alpha2-macroglobulin were examined in atopic subjects with common cold symptoms. Twenty-three patients with seasonal allergic rhinitis were inoculated with human rhinovirus 16 outside the pollen season. Diluent and allergen challenges, followed by nasal lavages, were carried out about 3 months before and 4 days after virus inoculation. Seventeen patients developed significant common cold symptoms with increased nasal lavage fluid levels of alpha2-macroglobulin, IL-8, and ECP at baseline (p<0.001-0.05 versus before inoculation), and were further increased by allergen challenge (p< 0.001-0.05); IL-8 and ECP levels were correlated (r = 0.63, p<0.001). Before inoculation, the six patients who later did not develop common cold symptoms had high levels of IL-8 and myeloperoxidase (MPO), and exhibited strong allergen-induced plasma exudation responses (alpha2-macroglobulin). After inoculation, IL-8 and ECP did not increase in these symptomless subjects. In conclusion, high nasal interleukin-8 and myeloperoxidase levels and exudative hyperresponsiveness may protect against infection. The association between nasal interleukin-8 and eosinophil cationic protein in common cold, particularly that observed in nasal lavage fluids after allergen-induced acute exudation of plasma, suggests the involvement of interleukin-8 in exacerbation of airway mucosal eosinophil activity.


title: Clathrin-dependent entry of severe acute respiratory syndrome coronavirus into target cells expressing ACE2 with the cytoplasmic tail deleted.

text: The penetration of various viruses into host cells is accomplished by hijacking the host endocytosis machinery. In the case of severe acute respiratory syndrome coronavirus (SARS-CoV) infection, viral entry is reported to require a low pH in intracytoplasmic vesicles; however, little is known about how SARS-CoV invades such compartments. Here we demonstrate that SARS-CoV mainly utilizes the clathrin-mediated endocytosis pathway for its entry to target cells by using infectious SARS-CoV, as well as a SARS-CoV pseudovirus packaged in the SARS-CoV envelope. The SARS-CoV entered caveolin-1-negative HepG2 cells, and the entry was significantly inhibited by treatment with chlorpromazine, an inhibitor for clathrin-dependent endocytosis, and by small interfering RNA-mediated gene silencing for the clathrin heavy chain. Furthermore, the SARS-CoV entered COS7 cells transfected with the mutant of ACE2 with the cytoplasmic tail deleted, SARS-CoV receptor, as well as the wild-type ACE2, and their entries were significantly inhibited by treatment with chlorpromazine. In addition, ACE2 translocated into EEA1-positive early endosomes immediately after the virus attachment to ACE2. These results suggest that when SARS-CoV binds ACE2 it is internalized and penetrates early endosomes in a clathrin-dependent manner and that the cytoplasmic tail of ACE2 is not required for the penetration of SARS-CoV.