title: Submaximal angioplasty for symptomatic intracranial atherosclerosis: a prospective Phase I study.

text: OBJECTIVE Intracranial atherosclerotic disease (ICAD) accounts for approximately 10% of ischemic strokes. The recent Stenting and Aggressive Medical Management for Preventing Recurrent Stroke in Intracranial Stenosis (SAMMPRIS) study demonstrated a high incidence of perioperative complications (15%) for treatment of ICAD with stenting. Although the incidence of stroke was lower in the medical arm, recurrent stroke was found in 12% of patients despite aggressive medical management, suggesting that intervention may remain a viable option for ICAD if perioperative risk is minimized. Angioplasty without stenting represents an alternative and understudied revascularization treatment for ICAD. Submaximal angioplasty limits the risks of thromboembolism, vessel perforation, and reperfusion hemorrhage that were frequently reported with stenting in the SAMMPRIS trial. The authors conducted a prospective Phase I trial designed to assess the safety of submaximal angioplasty in patients with symptomatic ICAD. METHODS This study was approved by the local institutional review board. Demographic and clinical data were prospectively collected. Angioplasty was performed with a balloon undersized to approximately 50%-70% of the nondiseased vessel diameter in patients with symptomatic ICAD who had angiographically significant stenosis of ≥ 70%. The primary outcome measure was the incidence of periprocedural complications (combined rate of death, stroke, and hemorrhage occurring within 30 days and at 1 year). RESULTS Among the 65 patients with symptomatic ICAD who were screened, 24 had significant angiographic stenosis that met the inclusion criteria of this study. The mean age was 64.08 years (median 65 years; SD ± 11.24 years), most were men (62.5%), and most were white (66.67%). Many patients had concomitants of vascular disease, including hypertension (95.8%), hyperlipidemia (70.83%), smoking history (54.1%), and diabetes mellitus (50.0%). Coronary artery disease (41.66%) and previous stroke or transient ischemic attack (45.83%) were frequently present. Most patients (75%) had anterior circulation stenosis. The mean preprocedure stenosis was 80.16% (median 80%, range 70%-95%). Submaximal angioplasty was performed in patients who met the inclusion criteria, with a mean postangioplasty stenosis rate of 54.62% (median 55.5%, range 31%-78%). Rates of ischemic stroke in the territory of the treated artery were 0% within 30 days and 5.55% (in the only patient who presented with recurrent stroke) at 1 year. The mortality and hemorrhage rates in this series were 0%. CONCLUSIONS This study demonstrates the safety of the submaximal angioplasty technique, with no permanent periprocedural complications in 24 treated patients.


title: Prospective Case Series of a Novel Minimally Invasive Bipolar Coagulation System in the Treatment of Grade I and II Internal Hemorrhoids.

text: Background Existing nonsurgical procedures for the treatment of grade I and II internal hemorrhoids are often painful, technically demanding, and often necessitate multiple applications. This study prospectively assessed the safety and efficacy of the HET Bipolar System, a novel minimally invasive device, in the treatment of symptomatic grade I and II internal hemorrhoids. Methods Patients with symptomatic grade I or II internal hemorrhoids despite medical management underwent hemorrhoidal ligation with the HET Bipolar System. Endpoints included resolution or improvement of hemorrhoidal bleeding and/or prolapse from baseline, recurrent or refractory symptoms, and pain. Results Twenty patients were treated with the HET Bipolar System. Two were lost to follow-up. Refractory or recurrent bleeding was present in 8 of 18 (44.4%), 4 of 11 (36.4%), and 4 of 8 (50.0%) patients, and prolapse was reported by 1 of 18 (5.6%), 4 of 11 (36.4%), and 1/7 (14.3%) of patients at 1, 3, and 6 months, respectively. Bleeding improved from baseline in 88.2%, 81.8%, and 87.5% of patients, and resolution of baseline prolapse was seen in 11 of 11 (100%), 4 of 7 (57.1%), and 5 of 5 (100%) patients at the same intervals. Thirteen of 18 (72.2%) patients did not require additional treatment for their symptoms. Conclusions The HET Bipolar System is safe and easy to use with short-term effectiveness comparable to that of currently used techniques for the treatment of symptomatic grade I and II internal hemorrhoids. It may be an effective alternative to rubber band ligation in patients with larger internal hemorrhoids and those with hemorrhoids close to the dentate line in which banding may produce debilitating pain.


title: Stabilization of a full-length infectious cDNA clone of transmissible gastroenteritis coronavirus by insertion of an intron.

text: The stable propagation of a full-length transmissible gastroenteritis coronavirus (TGEV) cDNA in Escherichia coli cells as a bacterial artificial chromosome has been considerably improved by the insertion of an intron to disrupt a toxic region identified in the viral genome. The viral RNA was expressed in the cell nucleus under the control of the cytomegalovirus promoter and the intron was efficiently removed during translocation of this RNA to the cytoplasm. The insertion in two different positions allowed stable plasmid amplification for at least 200 generations. Infectious TGEV was efficiently recovered from cells transfected with the modified cDNAs.


When viral oncoprotein meets tumor suppressor: a structural view.


title: New antimicrobial peptide active against Gram-positive pathogens.

text: BACKGROUND & OBJECTIVES Human and animal cystatins have been shown to inhibit the replication of certain viruses and bacteria, though it is not directly demonstrated that the effects are due to protease inhibitory capacity of the cystatins. We report antibacterial properties of a novel antimicrobial peptidyl derivative, (2S)-2-(N(alpha)-benzyloxycarbonyl-arginyl-leucylamido)-1-[(E)-cinnamoylamido]-3- methylbutane, structurally based upon the aminoterminal segment of the inhibitory centre of the human cysteine protease inhibitor, cystatin C. METHODS Clinical isolates of group A, B, C and G streptococci were collected. The antibacterial activity of Cystapep 1 derivative was tested by agar well diffusion method. RESULTS Cystapep 1, displayed antibacterial activity against several clinically important Gram-positive bacteria. It displayed minimal inhibitory and bactericidal concentrations of about 16 microg/ml for both Staphylococcus aureus and Streptococcus pyogenes. In radial agar diffusion assays, groups A, B, C and G streptococci as well as staphylococci were generally susceptible to the action of Cystapep 1, whereas pneumococci and enterococci were less susceptible. No activity against Gram-negative bacteria was observed. INTERPRETATION & CONCLUSION Cystapep 1 also showed high activity against methicillin-resistant Staph. aureus (MRSA) and multi-antibiotic resistant coagulase negative staphylococci (CNS), suggesting its mechanism of action to be different from most currently used antibiotics.


title: A field trial to evaluate the efficacy of a combined rotavirus-coronavirus/Escherichia coli vaccine in dairy cattle.

text: A field trial was designed to determine the efficacy of a combination rotavirus-coronavirus/Escherichia coli vaccine on dairy farms in southwestern Ontario. In Part A of the trial, 321 cows on 15 farms were randomly assigned to either vaccination or placebo groups. On eight farms, 50% of the dams were vaccinated, while on the other seven farms, 80% of the dams were vaccinated. In Part B of the trial, 26 farms were randomly assigned to either a total vaccination program or to no vaccination program. Mortality, disease occurrence and weight gains were recorded on all calves for the first two weeks of life. In Part A, 23.5% of all calves were treated in the first two weeks of life, 20.9% were treated specifically for scours and 3.6% of live-born calves died. Enteropathogenic E. coli was identified on 13 of the 15 farms, rotavirus on 11 and coronavirus on ten. At least one of the three potential pathogens was found on every farm. There were no significant differences between calves from placebo-treated and vaccine-treated dams with regard to the proportion treated for all diseases, or for scours, or the proportion which died. Neither were there differences in days to first treatment for all diseases (seven days on average), days to first scour (6.7 days), duration of treatments (3.9 days for all diseases, 3.7 days for scours), or estimated weight gains (0.5 kg/day to 14 days). These results were not altered when the presence or absence of enteropathogenic E. coli, rotavirus or coronavirus on the premises was accounted for.(ABSTRACT TRUNCATED AT 250 WORDS)


title: Molecular cloning and expression of a spike protein of neurovirulent murine coronavirus JHMV variant cl-2.

text: A cDNA encoding the spike (S) protein of the neurovirulent murine coronavirus JHMV variant cl-2 was isolated and sequenced. Analysis of the cDNA revealed that the S protein consists of 1376 amino acids, as does the S protein of mouse hepatitis virus 4. We inserted the cDNA into the genome of vaccinia virus to obtain a recombinant vaccinia virus (rVV). The S protein expressed in RK13 cells infected by the rVV was shown to be electrophoretically and immunologically indistinguishable from the S protein produced in DBT cells infected with cl-2 virus. RVV infection of rats and mice induced S protein-specific antibody production detectable by immunofluorescence and neutralization. Moreover, the S protein expressed by the rVV induced syncytium formation not only in mouse DBT and L cells, which are susceptible to cl-2 virus infection, but also in rabbit RK13 cells, which are not susceptible to cl-2 virus infection. This result suggests the possibility that RK13 cells have binding sites for the cl-2 virus S protein.


title: Amino acid sequence of a conserved neutralizing epitope of murine coronaviruses.

text: We identified the binding site of monoclonal antibody 19.2, which cross-neutralizes several mouse hepatitis virus (MHV) strains, inhibits fusion of MHV-infected cells, and protects against lethal infection (P. J. Talbot and M. J. Buchmeier, Virus Res. 2:317-328, 1985). We used fusion proteins, generated by expression of fragments of the MHV A59 E2 gene in pEX plasmids, and synthetic peptides in a PEPSCAN.


Tackling social determinants of health through community based initiatives.


Identifying hazards, assessing the risks.


Coronavirus antibodies in patients with multiple sclerosis.


Portable monitoring devices for diagnosis of obstructive sleep apnea at home: review of accuracy, cost-effectiveness, guidelines, and coverage in Canada.


title: Maturation and localization of macrophages and microglia during infection with a neurotropic murine coronavirus.

text: Macrophages and microglia are critical in the acute inflammatory response and act as final effector cells of demyelination during chronic infection with the neutrotropic MHV-JHM strain of mouse hepatitis virus (MHV-JHM). Herein, we show that "immature" F4/80(+)Ly-6C(hi) monocytes are the first cells, along with neutrophils, to enter the MHV-JHM-infected central nervous system (CNS). As the infection progresses, macrophages in the CNS down-regulate expression of Ly-6C and CD62L, consistent with maturation, and a higher frequency express CD11c, a marker for dendritic cells (DCs). Microglia also express CD11c during this phase of the infection. CD11c(+) macrophages in the infected CNS exhibit variable properties of immature antigen-presenting cells (APCs), with modestly increased CD40 and MHC expression, and equivalent potent antigen uptake when compared with CD11c(-) macrophages. Furthermore, CDllc(+) and F4/80(+) macrophages and microglia are localized to areas of demyelination, in some instances directly associated with damaged axons. These results suggest that chronic CNS infection results in the appearance of CD11c-expressing macrophages from the blood that exhibit properties of immature APCs, are closely associated with areas of demyelination, and may act as final effectors of myelin destruction.


title: Recapitulation of normal and abnormal BioBreeding rat T cell development in adult thymus organ culture.

text: Congenitally lymphopenic diabetes-prone (DP) BioBreeding (BB) rats develop spontaneous T cell-dependent autoimmunity. Coisogenic diabetes-resistant (DR) BB rats are not lymphopenic and are free of spontaneous autoimmune disease, but become diabetic in response to depletion of RT6+ T cells. The basis for the predisposition to autoimmunity in BB rats is unknown. Abnormal T cell development in DP-BB rats can be detected intrathymically, and thymocytes from DR-BB rats adoptively transfer diabetes. The mechanisms underlying these T cell developmental abnormalities are not known. To study these processes, we established adult thymus organ cultures (ATOC). We report that cultured DR- and DP-BB rat thymi generate mature CD4 and CD8 single-positive cells with up-regulated TCRs. DR-BB rat cultures also generate T cells that express RT6. In contrast, DP-BB rat cultures generate fewer CD4+, CD8+, and RT6+ T cells. Analysis of the cells obtained from ATOC suggested that the failure of cultured DP-BB rat thymi to generate T cells with a mature phenotype is due in part to an increased rate of apoptosis. Consistent with this inference, we observed that addition of the general caspase inhibitor Z-VAD-FMK substantially increases the number of both mature and immature T cells produced by DP-BB rat ATOC. We conclude that cultured DR-BB and DP-BB rat thymi, respectively, recapitulate the normal and abnormal T cell developmental kinetics and phenotypes observed in these animals in vivo. Such cultures should facilitate identification of the underlying pathological processes that lead to immune dysfunction and autoimmunity in BB rats.


title: [Robotic and laparosocpic urological surgery during COVID-19 pandemia.]

text: OBJECTIVE SARS-CoV-2 pandemic hashigh repercussion on urologic minimally invasive surgery (MIS). Controversy about safety of MIS procedures during COVID-19 pandemic has been published. Nowadays, our priority should be create agreement in order to restart and organize MIS with safety conditions for patients and healthcare workers. METHODS: Pubmed and web search was conducted with following terms: "SARS-CoV-2", "COVID19", "COVID19 Urology", COVID19 Surgery", "COVID19 transmission", "SARS-CoV-2 transmission", "COVID19 nd minimally invasive surgery", "SARS-CoV-2 and CO 2insuflation". A narrative review of available literature and scientific evidence summary was done. A modify nominal group technique was used to achieve an expert consensus. First draft was circulated amongst authors. Definitive document was approved in May 26th. RESULTS: Non evidence supports higher risk of SARSCoV-2 healthcare workers infection with MIS compared to open surgery. MIS is associated with shorter hospital stay than open surgery. Modify MIS indications to open surgery, with no scientific evidence, could spend valuable resources in detriment to COVID-19 patients. MIS indications should be prioritized attending to available resources and pandemic intensity. SARS-CoV-2screening 72 hours prior to surgery by clinical and epidemiological questionnaire and nasopharyngeal PCRis recommended, in order to prevent nosocomial transmission, professional infections and to minimize postoperative complications. Intraoperative steps should be established to reduce professional exposure to surgical aerosols, including: surgical room reorganization, adequate personal protective equipment, surgical technique optimization and management of CO2 and surgical smoke. CONCLUSIONS In COVID-19 pandemic de-escalation, MIS carried out with optimal safety measurements, could contribute to reduce hospital resources utilization. With current evidence, MIS should not be limited or reconverted to open surgery during COVID-19 pandemic.


title: Anesthesia for the child with an upper respiratory tract infection: still a dilemma?

text: One of the most controversial issues in pediatric anesthesia has revolved around the decision to proceed with anesthesia and surgery for the child who presents with an upper respiratory tract infection (URI). In the past, doctrine dictated that children with URIs have their surgery postponed until the child was symptom free. This practice was based on the empirically supported premise that anesthesia increased the risk of serious complications and complicated the child's postoperative course. Although recent clinical data confirm that some children with URIs are at increased risk of perioperative complications, these complications can, for the most part, be anticipated, recognized, and treated. Although the child with a URI still presents a challenge, anesthesiologists are now in a better position to make informed decisions regarding the assessment and management of these children, such that blanket cancellation has now become a thing of the past.


title: The role of superspreading in Middle East respiratory syndrome coronavirus (MERS-CoV) transmission.

text: As at 15 June 2015, a large transmission cluster of Middle East respiratory syndrome coronavirus (MERSCoV)was ongoing in South Korea. To examine the potential for such events, we estimated the level of heterogeneity in MERS-CoV transmission by analyzing data on cluster size distributions. We found substantial potential for superspreading; even though it is likely that R0 < 1 overall, our analysis indicates that cluster sizes of over 150 cases are not unexpected forMERS-CoV infection.


title: Catheter-based renal denervation for treatment of resistant hypertension.

text: INTRODUCTION Activation of renal sympathetic nerves is associated with the development of hypertension. Catheter-based renal sympathetic denervation with radiofrequency energy ablation is a new promising treatment option for resistant hypertension. We here report the first Danish experiences and results with this technique. MATERIAL AND METHODS Nine patients with resistant hypertension and a day-time 24-hour ambulatory blood pressure (BP) of 152/89 mmHg ± 10/10 (standard deviation) mmHg despite treatment with 5.4 ± 1.4 anti-hypertensive drugs underwent catheter-based renal sympathetic denervation with the Symplicity catheter. RESULTS No periprocedural complications or adverse events during follow-up were observed. Seven patients received complete ablation and two patients only partial ablation. Five patients responded to the treatment with a reduction in day-time 24-hour ambulatory BP from 158/94 ± 13/9 mmHg to 139/82 ± 10/8 mmHg (p < 0.05) at the one month follow-up and a reduction in the number of anti-hypertensive drugs from 5.4 ± 1.6 to 3.4 ± 0.9 (p < 0.05). BP in the remaining four patients was not significantly changed and antihypertensive therapy was not changed. CONCLUSION Catheter-based renal sympathetic denervation is a feasible and in several cases also effective treatment option for patients with resistant hypertension. Adequately designed controlled trials are needed to assess the long-term safety and the full potential of this treatment.


title: Cellular origin and ultrastructure of membranes induced during poliovirus infection.

text: Poliovirus RNA replicative complexes are associated with cytoplasmic membranous structures that accumulate during viral infection. These membranes were immunoisolated by using a monoclonal antibody against the viral nonstructural protein 2C. Biochemical analysis of the isolated membranes revealed that several organelles of the host cell (lysosomes, trans-Golgi stack and trans-Golgi network, and endoplasmic reticulum) contributed to the virus-induced membranous structures. Electron microscopy of infected cells preserved by high-pressure freezing revealed that the virus-induced membranes contain double lipid bilayers that surround apparently cytosolic material. Immunolabeling experiments showed that poliovirus proteins 2C and 3D were localized to the same membranes as the cellular markers tested. The morphological and biochemical data are consistent with the hypothesis that autophagy or a similar host process is involved in the formation of the poliovirus-induced membranes.


Open-access chemistry databases evolving slowly but not surely.


title: Comparative study of the effects of heptameric slippery site composition on -1 frameshifting among different eukaryotic systems.

text: Studies of programmed -1 ribosomal frameshifting (-1 PRF) have been approached over the past two decades by many different laboratories using a diverse array of virus-derived frameshift signals in translational assay systems derived from a variety of sources. Though it is generally acknowledged that both absolute and relative -1 PRF efficiency can vary in an assay system-dependent manner, no methodical study of this phenomenon has been undertaken. To address this issue, a series of slippery site mutants of the SARS-associated coronavirus frameshift signal were systematically assayed in four different eukaryotic translational systems. HIV-1 promoted frameshifting was also compared between Escherichia coli and a human T-cell line expression systems. The results of these analyses highlight different aspects of each system, suggesting in general that (1) differences can be due to the assay systems themselves; (2) phylogenetic differences in ribosome structure can affect frameshifting efficiency; and (3) care must be taken to employ the closest phylogenetic match between a specific -1 PRF signal and the choice of translational assay system.


title: Development of a focus-reduction neutralizing test for detecting equine herpesvirus type-1-neutralizing antibodies.

text: Virus-neutralizing (VN) testing is essential for evaluating virus-specific immunity in equine herpesvirus type-1 (EHV-1) infection. We developed a focus-reduction neutralization test (FRNT) for EHV-1 using 96-well plates for faster large-scale testing with sufficient sensitivity. We used an overlay medium containing Avicel (FMC Biopolymer), a microcrystalline cellulose with lower viscosity than the methylcellulose. The foci were visualized by immuno-staining with anti-EHV-1 gp14 monoclonal antibody. The FRNT successfully detected seroconversion in horses experimentally infected with EHV-1 (n = 3) and in those infected naturally (n = 16). The FRNT for EHV-1 was high-throughput and time saving. The FRNT can be used in large-scale seroepidemiological studies of EHV-1 and in evaluating vaccine efficacy.


title: Computational methods for remote homolog identification.

text: As more and more protein sequences are available, homolog identification becomes increasingly important for functional, structural, and evolutional studies of proteins. Many homologous proteins were separated a very long time ago in their evolutionary history and thus their sequences share low sequence identity. These remote homologs have become a research focus in bioinformatics over the past decade, and some significant advances have been achieved. In this paper, we provide a comprehensive review on computational techniques used in remote homolog identification based on different methods, including sequence-sequence comparison, and sequence-structure comparison, and structure-structure comparison. Other miscellaneous approaches are also summarized. Pointers to the online resources of these methods and their related databases are provided. Comparisons among different methods in terms of their technical approaches, their strengths, and limitations are followed. Studies on proteins in SARS-CoV are shown as an example for remote homolog identification application.


title: Canine viruses and multiple sclerosis.

text: Serum and cerebrospinal fluid from multiple sclerosis (MS) patients and control subjects were tested and compared for presence and titer of neutralizing antibody against the most common canine viruses. Canine viruses included canine distemper virus (CDV), canine adenovirus 1 (CAV-1), canine parainfluenza virus (CPIV), canine herpesvirus (CHV), canine coronavirus (CCV), and canine parvovirus (CPV). Neutralizing titers against measles virus (MV) and human adenovirus 8 (HA8) were also tested. Significantly elevated (p less than 0.05) antibody levels in sera from MS patients were found only against MV and CDV, but this depended upon the study population and the method of evaluation. The CDV-neutralizing component in serum could be absorbed on MV-infected cells. Results of this study failed to establish a link between canine viruses and MS.


title: Emotional and behavioral consequences of bioterrorism: planning a public health response.

text: Millions of dollars have been spent improving the public health system's bioterrorism response capabilities. Yet relatively little attention has been paid to precisely how the public will respond to bioterrorism and how emotional and behavioral responses might complicate an otherwise successful response. This article synthesizes the available evidence about the likely emotional and behavioral consequences of bioterrorism to suggest what decision makers can do now to improve that response. It examines the emotional and behavioral impact of previous "bioterrorism-like" events and summarizes interviews with experts who have responded to such events or conducted research on the effects of community-wide disasters. The article concludes by reflecting on the evidence and experts' perspectives to suggest actions to be taken now and future policy and research priorities.


title: Prevalence of rotavirus and coronavirus antigens in the feces of normal cows.

text: The prevalence of rotavirus and coronavirus shedding by adult cows was investigated using capture enzyme-linked immunosorbent assays. Fecal samples from 121 cows in a single herd were tested for the presence of rotavirus and coronavirus, either free or complexed with immunoglobulin. Free rotavirus was not detected in any samples while rotavirus-immunoglobulin complexes were detected in 53 of 121 (44%) samples tested. In contrast, free coronavirus was detected in six (5%) samples and coronavirus-immunoglobulin complexes were detected in 85 (70%) of the samples tested. Thus it appears that subclinical infection of cows by either of these viruses is common, possibly providing a source for infection of the neonate. These assays may therefore provide important information regarding the epidemiology of enteric virus infections and suggest means of improving management to prevent epidemics of neonatal diarrhea.


title: CCR7 is critically important for migration of dendritic cells in intestinal lamina propria to mesenteric lymph nodes.

text: Although dendritic cells (DCs) located in the small intestinal lamina propria (LP-DCs) migrate to mesenteric lymph nodes (MLNs) constitutively, it is unclear which chemokines regulate their trafficking to MLNs. In this study we report that LP-DCs in unperturbed mice require CCR7 to migrate to MLNs. In vitro, LP-DCs expressing CCR7 migrated toward CCL21, although the LP-DCs appeared morphologically and phenotypically immature. In MLNs, DCs bearing the unique LP-DC phenotype (CD11chighCD8alphaintCD11blowalphaLlowbeta7high and CD11chighCD8alpha-CD11bhighalphaLlowbeta7high) were abundant in wild-type mice, but were markedly fewer in CCL19-, CCL21-Ser-deficient plt/plt mice and were almost absent in CCR7-deficient mice, indicating the critical importance of CCR7 in LP-DC trafficking to MLNs. Interestingly, CCR7+ DCs in MLNs with the unique LP-DC phenotype had numerous vacuoles containing cellular debris in the cytoplasm, although MLN-DCs themselves were poorly phagocytic, suggesting that the debris was derived from the LP, where the LP-DCs ingested apoptotic intestinal epithelial cells (IECs). Consistent with this, LP-DCs ingested IECs vigorously in vitro. By presenting IEC-associated Ag, the LP-DCs also induce T cells to produce IL-4 and IL-10. Collectively, these results strongly suggest that LP-DCs with unique immunomodulatory activities migrate to MLNs in a CCR7-dependent manner to engage in the presentation of IEC-associated Ags acquired in the LP.


title: [Detection of Middle East Respiratory Syndrome Coronavirus by Reverse-transcription Loop-Mediated Isothermal Amplification].

text: A simple, rapid and sensitive colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) was developed for rapid detection of Middle East respiratory syndrome coronavirus (MERS-CoV). The method employed six primers that recognized sequences of a nucleocapsid gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 60 min. Products were detected through a LA-320c Loopamp Turbidimeter (real-time RT-LAMP) or visual inspection of color change by pre-addition of Hydroxynaphthol Blue dye (visual RT-LAMP). Specificity of RT-LAMP was validated by detection of several human coronaviruses and common respiratory viruses. MERS-CoV real-time RT-LAMP had a linear correlation (R2) of 0.995 at 10(3)-10(6) copies. The limit of detection of real-time RT-LAMP, visual RT-LAMP and quantitative real-time PCR was 500, 1000 and 100 copies/reaction, respectively. The established RT-LAMP assay was demonstrated to be a rapid screening tool for MERS-CoV infection, and could be suitable in resource-limited clinical sites and for field studies.


title: Application of RT-Bst to enhance detection of pathogenic viruses of the respiratory tract.

text: Inefficiency of RT-PCR can be associated with the suboptimal process of reverse transcription as only 40-80% of RNA is converted to cDNA. We employed a novel method, RT-Bst, to enrich the concentration of cDNA for subsequent multiplex PCR detection of selected RNA viruses. The RT-Bst method amplifies cDNA through reverse transcription of viral RNA using reverse transcriptase and amplification of cDNA using Bst DNA polymerase. Viral RNA was extracted from 25 nasopharyngeal samples for detection of influenza A, B and C; parainfluenza 1-4; human coronaviruses 229E and OC43; respiratory syncytial virus (RSV) and rhinovirus. Both multiplex one-step RT-PCR and RT-Bst PCR were used to compare their performances for detection of virus sequences. These findings were compared with routine laboratory detection. When using RT-Bst PCR, 28% of samples yielded a viral pathogen compared to 20% with RT-PCR and 12% using routine diagnostic tests. RT-Bst PCR was shown to have particular utility in the detection of RSV RNA as this was present in 20% of the samples studied compared to 8% when using RT-PCR. For one patient, RT-Bst PCR was able to detect RSV five days earlier than conventional hospital diagnostic testing. RT-Bst and RT-Bst PCR can be used as alternative approaches to reverse transcription and one-step RT-PCR, respectively, for sequence-independent amplification of RNA virus sequences and a larger scale analysis of this new diagnostic approach is warranted.


title: Vaccines for the prevention of respiratory viral infections: problems and current status.

text: Acute respiratory virus infections cause the majority of lower respiratory tract illnesses and hospitalisations of infants and the elderly. The emergence of new respiratory viruses and a high probability that influenza will cause further pandemics highlights the necessity for developing better preventative strategies. Although there is a clear and pressing need for vaccines to prevent respiratory syncytial virus, rhinoviruses, coronaviruses, parainfluenza and human metapneumovirus, progress has been extremely slow. This review presents the current status of vaccine development for respiratory viral diseases and outlines novel approaches for the future.


title: Correlation between blood and lymphatic vessel density and results of contrast-enhanced spectral mammography.

text: UNLABELLED Contrast-enhanced spectral mammography (CESM) is a novel technique used for detection of tumour vascularity by imaging the moment in which contrast, delivered to the lesion by blood vessels, leaks out of them, and flows out through lymphatic vessels. In our study, we included 174 women for whom spectral mammography was performed for diagnostic purposes. The relationship between enhancement in CESM and blood vessel density (BVD), lymphatic vessel density (LVD) or the percentage of fields with at least one lymphatic vessel (distribution of podoplanin-positive vessels - DPV) and other related parameters was assessed in 55 cases. BVD, LVD and DPV were assessed immunohistochemically, applying podoplanin and CD31/CD34 as markers of lymphatic and blood vessels, respectively. The sensitivity (in detection of malignant lesions) of CESM was 100%, while its specificity - 39%. We found a significant positive correlation between the intensity of enhancement in CESM and BVD (p = 0.007, r = 0.357) and a negative correlation between the intensity of enhancement in CESM and DPV (p = 0.003, r = -0.390). Lesions with the highest enhancement in CESM showed a high number of blood vessels and a low number of lymphatics. CONCLUSIONS 1) CESM is a method characterized by high sensitivity and acceptable specificity; 2) the correlation between CESM results and blood/lymphatic vessel density confirms its utility in detection of tissue angiogenesis and/or lymphangiogenesis.


title: Biochemical aspects of coronavirus replication and virus-host interaction.

text: Infection by different coronaviruses (CoVs) causes alterations in the transcriptional and translational patterns, cell cycle, cytoskeleton, and apoptosis pathways of the host cells. In addition, CoV infection may cause inflammation, alter immune and stress responses, and modify the coagulation pathways. The balance between the up- and downregulated genes could explain the pathogenesis caused by these viruses. We review specific aspects of CoV-host interactions. CoV genome replication takes place in the cytoplasm in a membrane-protected microenvironment and may control the cell machinery by locating some of their proteins in the host cell nucleus. CoVs initiate translation by cap-dependent and cap-independent mechanisms. CoV transcription involves a discontinuous RNA synthesis (template switching) during the extension of a negative copy of the subgenomic mRNAs. The requirement for base-pairing during transcription has been formally demonstrated in arteriviruses and CoVs. CoV N proteins have RNA chaperone activity that may help initiate template switching. Both viral and cellular proteins are required for replication and transcription, and the role of selected proteins is addressed.


title: Arterial aneurysms associated with arteriovenous malformations of the brain: classification, incidence, risk of hemorrhage, and treatment-a systematic review.

text: BACKGROUND Aneurysms associated with brain arteriovenous malformations (bAVMs) influence the natural history of these lesions and pose important therapeutic challenges. However, the epidemiology, natural history, and appropriate management of the aneurysms associated with bAVMs are not completely understood due to the paucity of large and uniform studies. We performed a systematic review of published series examining the association between aneurysms and bAVMs with the purpose of clarifying the prevalence, risk of hemorrhage, and appropriate management of these lesions. METHOD PRISMA/MOOSE guidelines were followed. We conducted a comprehensive literature search of three databases (PubMed, Ovid MEDLINE, and Ovid EMBASE) on aneurysms associated with bAVMs. Only studies examining consecutive case series of aneurysms associated with bAVMs were included. From the collected studies, we extracted data regarding prevalence of bAVM-associated aneurysms, risk of aneurysm rupture in relation to bAVM location and aneurysm characteristics, and treatment-related outcomes. RESULTS Our systematic review included 44 articles with a total of 10,093 bAVMs. The proportion of bAVMs with an associated aneurysm was 20.2 % (95 % CI = 19.4-20.9 %). Among ruptured bAVMs with associated aneurysms, the aneurysm was the source of hemorrhage in 49.2 % (95 % CI = 43.7-54.7 %) of cases. Flow-related aneurysms were the most common source of aneurysm rupture (78.5 %, 95 % CI = 70.6-84.9 %). Infratentorial bAVM-associated aneurysms presented a higher risk of rupture (60 %, 95 % CI = 47.4-71.9 %) when compared with supratentorial lesions (29 %, 95 % CI = 21.4-38.5 %). Endovascular treatment of aneurysms associated with bAVMs had a cure rate of 80.0 % (95 % CI = 73.3-85.3 %), complication rate of 8.7 % (95 % CI = 5.5-13.1 %), and a good neurological outcome rate of 78.8 % (95 % CI = 72.5-83.9 %). CONCLUSIONS Twenty percent of bAVMs harbored arterial aneurysms. The presence of aneurysm increases the risk of bleeding of the bAVM, especially when flow-related or infratentorially located. Aneurysms associated with bAVMs should be treated promptly. Selective endovascular treatment of bAVM-associated aneurysms appears safe and effective.


title: Carboxyl terminus of severe acute respiratory syndrome coronavirus nucleocapsid protein: self-association analysis and nucleic acid binding characterization.

text: Coronavirus nucleocapsid (N) protein envelops the genomic RNA to form long helical nucleocapsid during virion assembly. Since N protein oligomerization is usually a crucial step in this process, characterization of such an oligomerization will help in the understanding of the possible mechanisms for nucleocapsid formation. The N protein of severe acute respiratory syndrome coronavirus (SARS-CoV) was recently discovered to self-associate by its carboxyl terminus. In this study, to further address the detailed understanding of the association feature of this C-terminus, its oligomerization was systematically investigated by size exclusion chromatography and chemical cross-linking assays. Our results clearly indicated that the C-terminal domain of SARS-CoV N protein could form not only dimers but also trimers, tetramers, and hexamers. Further analyses against six deletion mutants showed that residues 343-402 were necessary and sufficient for this C-terminus oligomerization. Although this segment contains many charged residues, differences in ionic strength have no effects on its oligomerization, indicating the absence of electrostatic force in SARS-CoV N protein C-terminus self-association. Gel shift assay results revealed that the SARS-CoV N protein C-terminus is also able to associate with nucleic acids and residues 363-382 are the responsible interaction partner, demonstrating that this fragment might involve genomic RNA binding sites. The fact that nucleic acid binding could promote the SARS-CoV N protein C-terminus to form high-order oligomers implies that the oligomeric SARS-CoV N protein probably combines with the viral genomic RNA in triggering long nucleocapsid formation.


title: Quantitative systematic review of randomised controlled trials comparing antibiotic with placebo for acute cough in adults.

text: OBJECTIVES To assess whether antibiotic treatment for acute cough is effective and to measure the side effects of such treatment. DESIGN Quantitative systematic review of randomised placebo controlled trials. DATA SOURCES Nine trials (8 published, 1 unpublished) retrieved from a systematic search (electronic databases, contact with authors, contact with drug manufacturers, reference lists); no restriction on language. MAIN OUTCOME MEASURES Proportion of subjects with productive cough at follow up (7-11 days after consultation with general practitioner); proportion of subjects who had not improved clinically at follow up; proportion of subjects who reported side effects from taking antibiotic or placebo. RESULTS Eight trials contributed to the meta-analysis. Resolution of cough was not affected by antibiotic treatment (relative risk 0.85 (95% confidence interval 0.73 to 1.00)), neither was clinical improvement at re-examination (relative risk 0.62 (0.36 to 1.09)). The side effects of antibiotic were more common in the antibiotic group when compared to placebo (relative risk 1.51 (0.86 to 2.64)). CONCLUSIONS Treatment with antibiotic does not affect the resolution of cough or alter the course of illness. The benefits of antibiotic treatment are marginal for most patients with acute cough and may be outweighed by the side effects of treatment.


title: Daily Situation Report on Coronavirus disease (COVID-19) in Iran; March 22, 2020.

text: To forecast the COVID-19 epidemic trend in Iran, a set of dynamic models were created. In these models, the effect of climate and community behavior change on the reproductive number is modeled. A baseline scenario is also defined as letting the epidemic grow naturally, with no prevention/control interventions. Different levels of intervention are also defined based on the extent of population isolation. Outcomes of each scenario on the number of infected cases are computed. Modeling results shows that if effective isolation occurs in 10% of the population, 307,000 individuals will be infected with COVID-19 in Iran by May 20, 2020. For an isolation rate of 25%, started since March 10, 2020, cumulative number of deaths will be 13,450 individuals by May 20, 2020. If preventive efforts can reach a 32% effective isolation rate, a considerable decrease in the number of new cases would be expected during April and May, and the total number of deaths would decrease to 8,630. If the isolation rate increases to 40%, the total number of infected individuals is expected to be 6,030 individuals. It should be acknowledged that this model predicts the number of deaths of all COVID-19 cases, including those who will not be confirmed as COVID-19, because of non-testing or death prior to COVID-19 identification. This serious health issue requires both long-term and short-term planning and arrangements. Social distancing, and identification and isolation of suspected and confirmed cases are believed to be the most important and effective control measure, and should be strengthened soon. For further details on the methodology and results of these models, please visit: http://corona.behdasht.gov.ir/files/site1/files/Covid_Modeling_V14_26.12.98.pdf.


Will travel restrictions control the international spread of pandemic influenza?


title: Murine coronavirus replication induces cell cycle arrest in G0/G1 phase.

text: Mouse hepatitis virus (MHV) replication in actively growing DBT and 17Cl-1 cells resulted in the inhibition of host cellular DNA synthesis and the accumulation of infected cells in the G(0)/G(1) phase of the cell cycle. UV-irradiated MHV failed to inhibit host cellular DNA synthesis. MHV infection in quiescent 17Cl-1 cells that had been synchronized in the G(0) phase by serum deprivation prevented infected cells from entering the S phase after serum stimulation. MHV replication inhibited hyperphosphorylation of the retinoblastoma protein (pRb), the event that is necessary for cell cycle progression through late G(1) and into the S phase. While the amounts of the cellular cyclin-dependent kinase (Cdk) inhibitors p21(Cip1), p27(Kip1), and p16(INK4a) did not change in infected cells, MHV infection in asynchronous cultures induced a clear reduction in the amounts of Cdk4 and G(1) cyclins (cyclins D1, D2, D3, and E) in both DBT and 17Cl-1 cells and a reduction in Cdk6 levels in 17Cl-1 cells. Infection also resulted in a decrease in Cdk2 activity in both cell lines. MHV infection in quiescent 17Cl-1 cells prevented normal increases in Cdk4, Cdk6, cyclin D1, and cyclin D3 levels after serum stimulation. The amounts of cyclin D2 and cyclin E were not increased significantly after serum stimulation in mock-infected cells, whereas they were decreased in MHV-infected cells, suggesting the possibility that MHV infection may induce cyclin D2 and cyclin E degradation. Our data suggested that a reduction in the amounts of G(1) cyclin-Cdk complexes in MHV-infected cells led to a reduction in Cdk activities and insufficient hyperphosphorylation of pRb, resulting in inhibition of the cell cycle in the G(0)/G(1) phase.


Letter by Venema et al Regarding Article, "Analysis of Workflow and Time to Treatment on Thrombectomy Outcome in the Endovascular Treatment for Small Core and Proximal Occlusion Ischemic Stroke (ESCAPE) Randomized, Controlled Trial".


title: [Severe acute respiratory syndrome epidemiology in Sichuan province].

text: OBJECTIVE To explore the epidemiological characters of severe acute respiratory syndrome (SARS) in Sichuan province in order to provide evidence for prevention and control. METHODS To generate data on SARS in Sichuan province through descriptive and analytical studies on time, geographic distribution, population, source of infection, the way of case finding, symptom, diagnosis and treatment of the cases. RESULTS The peak of the epidemic last from April 16 to May 7. The number of cases in Luzhou and Guangyuan cities took up 60% of the total. Mobile population occupied 68% of the cases. Most of the patients were above the age of 20 with a sex ratio of 1.5:1 (m/f). 80% of the cases had a history of working in Guangdong province and recently returned to their hometowns. The main symptoms and signs of the SARS patients would include fever, cough and chest X-ray changes. CONCLUSION All cases were imported. Fluctuation of the epidemics was mainly affected by the mobility of working population who recently returned to their hometowns. Measures concerning the prevention and control of the epidemics would mainly target on the isolation of confirmed and suspected patients who might serve as the sources of infection through setting up quarantine station, assigned hospitals and special 'fever-clinics'.


title: Stability of SARS coronavirus in human specimens and environment and its sensitivity to heating and UV irradiation.

text: OBJECTIVE The causal agent for SARS is considered as a novel coronavirus that has never been described both in human and animals previously. The stability of SARS coronavirus in human specimens and in environments was studied. METHODS Using a SARS coronavirus strain CoV-P9, which was isolated from pharyngeal swab of a probable SARS case in Beijing, its stability in mimic human specimens and in mimic environment including surfaces of commonly used materials or in household conditions, as well as its resistance to temperature and UV irradiation were analyzed. A total of 10(6) TCID50 viruses were placed in each tested condition, and changes of the viral infectivity in samples after treatments were measured by evaluating cytopathic effect (CPE) in cell line Vero-E6 at 48 h after infection. RESULTS The results showed that SARS coronavirus in the testing condition could survive in serum, 1:20 diluted sputum and feces for at least 96 h, whereas it could remain alive in urine for at least 72 h with a low level of infectivity. The survival abilities on the surfaces of eight different materials and in water were quite comparable, revealing reduction of infectivity after 72 to 96 h exposure. Viruses stayed stable at 4 degrees C, at room temperature (20 degrees C) and at 37 degrees C for at least 2 h without remarkable change in the infectious ability in cells, but were converted to be non-infectious after 90-, 60- and 30-min exposure at 56 degrees C, at 67 degrees C and at 75 degrees C, respectively. Irradiation of UV for 60 min on the virus in culture medium resulted in the destruction of viral infectivity at an undetectable level. CONCLUSION The survival ability of SARS coronavirus in human specimens and in environments seems to be relatively strong. Heating and UV irradiation can efficiently eliminate the viral infectivity.


title: Leukocyte-specific protein 1 interacts with DC-SIGN and mediates transport of HIV to the proteasome in dendritic cells.

text: Dendritic cells (DCs) capture and internalize human immunodeficiency virus (HIV)-1 through C-type lectins, including DC-SIGN. These cells mediate efficient infection of T cells by concentrating the delivery of virus through the infectious synapse, a process dependent on the cytoplasmic domain of DC-SIGN. Here, we identify a cellular protein that binds specifically to the cytoplasmic region of DC-SIGN and directs internalized virus to the proteasome. This cellular protein, leukocyte-specific protein 1 (LSP1), was defined biochemically by immunoprecipitation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. LSP1 is an F-actin binding protein involved in leukocyte motility and found on the cytoplasmic surface of the plasma membrane. LSP1 interacted specifically with DC-SIGN and other C-type lectins, but not the inactive mutant DC-SIGNDelta35, which lacks a cytoplasmic domain and shows altered virus transport in DCs. LSP1 diverts HIV-1 to the proteasome. Down-regulation of LSP1 with specific small interfering RNAs in human DCs enhanced HIV-1 transfer to T cells, and bone marrow DCs from lsp1(-/-) mice also showed an increase in transfer of HIV-1(BaL) to a human T cell line. Proteasome inhibitors increased retention of viral proteins in lsp1(+/+) DCs, and substantial colocalization of virus to the proteasome was observed in wild-type compared with LSP1-deficient cells. Collectively, these data suggest that LSP1 protein facilitates virus transport into the proteasome after its interaction with DC-SIGN through its interaction with cytoskeletal proteins.


title: An initial investigation of the association between the SARS outbreak and weather: with the view of the environmental temperature and its variation.

text: OBJECTIVE To understand the association between the SARS outbreak and the environmental temperature, and to provide a scientific basis for prevention and control measures against it. METHODS The daily numbers of the probable SARS patients and the daily meteorological factors during the SARS outbreak period in Hong Kong, Guangzhou, Beijing, and Taiyuan were used in the data analysis. Ecological analysis was conducted to explore the association between the daily numbers of probable SARS patients and the environmental temperature and its variations. RESULTS There was a significant correlation between the SARS cases and the environmental temperature seven days before the onset and the seven day time lag corresponds well with the known incubation period for SARS. The optimum environmental temperature associated with the SARS cases was between 16 degrees C to 28 degrees C, which may encourage virus growth. A sharp rise or decrease in the environmental temperature related to the cold spell led to an increase of the SARS cases because of the possible influence of the weather on the human immune system. This study provided some evidence that there is a higher possibility for SARS to reoccur in spring than that in autumn and winter. CONCLUSION Current knowledge based on case studies of the SARS outbreak in the four cities suggested that the SARS outbreaks were significantly associated with the temperature and its variations. However, because the fallacy and the uncontrolled confounding effects might have biased the results, the possibility of other meteorological factors having an affect on the SARS outbreaks deserves further investigation.


title: The prevalence of types I and II feline coronavirus infections in cats.

text: The types of feline coronaviruses that are prevalent throughout Japan were determined by competitive enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody (MAb) to feline infectious peritonitis virus (FIPV) Type II and neutralizing test using Type II FIPV as challenge virus. A total of 1,079 cat serum samples were tested by indirect fluorescent antibody (IFA) assay for FIPV Type II antigen, all 42 sample from natural cases of FIP, 138 of 647 (21.3%) from cases with some chronic diseases and 57 of 390 (14.6%) from apparently non-diseased cases were positive. Of the 42 cases with FIP, 29 (69%) and 13 (31%) were found to have infection with FIPV Types I and II, respectively. Of the cases with chronic diseases, 111 (80.4%) were shown to have infection with FIPV or FECV Type I, while 14 (10.1%) with FIPV or FECV Type II. All of the 57 apparently non-diseased cases seemed to have been infected with FIPV or FECV Type I. These results indicated that feline coronavirus Type I is more high prevalent in Japan.


title: Severe acute respiratory syndrome: correlation between clinical outcome and radiologic features.

text: PURPOSE To evaluate whether there is a correlation between the clinical outcomes and radiologic features of severe acute respiratory syndrome (SARS). MATERIALS AND METHODS The clinical, laboratory, and radiologic features of 138 patients with SARS were analyzed. Three radiologists in consensus retrospectively assessed the frontal chest radiographs obtained at presentation and during treatment (n = 2045) for the distribution (each lung was divided into upper, middle, and lower zones) and extent of lung parenchymal abnormality. Clinical end points included intensive care unit (ICU) admission and death. RESULTS Thirty-six (26.1%) patients required ICU care, and eight (5.8%) died. The patients who required ICU care and/or died had more extensive consolidation on chest radiographs obtained initially (median percentage of consolidation, 3.30%, with interquartile range [IR] of 1.70%-8.78% vs 1.70% [IR, 0%-3.30%]; P < .001) and on day 7 after fever onset (median percentage of consolidation, 15.00% [IR, 6.48%-28.73%] vs 5.00% [IR, 2.50%-7.50%]; P < .001) than did surviving patients who did not require ICU care. Patients with involvement of more than one lung zone on initial and day 7 chest radiographs were more likely to require ICU care and/or die than were those with involvement of one or fewer zones (P < .001). Patients with bilateral pneumonic changes at presentation were more likely to have an adverse outcome than were those with unilateral pneumonia (P < .001). Involvement of more than one lung zone at baseline chest radiography was an independent predictor of ICU admission and/or death (odds ratio, 3.16; 95% confidence interval: 1.07, 9.32; P = .037) after adjustments for other significant factors (ie, patient age, and baseline neutrophil count and lactate dehydrogenase level). CONCLUSION More extensive airspace disease at presentation is an independent predictor of adverse outcome in patients with SARS.


title: Detection of rodent coronaviruses by use of fluorogenic reverse transcriptase-polymerase chain reaction analysis.

text: Reverse transcriptase-polymerase chain reaction (RT-PCR) assays have proved useful for the detection of mouse hepatitis virus (MHV) and rat coronavirus (RCV) in acutely infected animals and contaminated biomaterials. Fluorogenic nuclease RT-PCR assays combine RT-PCR with an internal fluorogenic hybridization probe, thereby eliminating post-PCR processing and potentially enhancing specificity. Consequently, a fluorogenic nuclease RT-PCR assay specific for rodent coronaviruses was developed. Primer and probe sequences were selected from the viral genome segment that encodes the membrane (M) protein that is highly conserved among rodent coronaviruses. Use of the fluorogenic nuclease RT-PCR detected all strains of MHV and RCV that were evaluated, but did not detect other RNA viruses that naturally infect rodents. Use of the assay detected as little as two femtograms of in vitro transcribed RNA generated from cloned amplicon, and when compared directly with mouse antibody production tests, had similar sensitivity at detecting MHV-A59 in infected cell culture lysates. Finally, use of the assay detected coronavirus RNA in tissues, cage swipes, and feces obtained from mice experimentally infected with MHV, and in tissues and cage swipes obtained from rats naturally infected with RCV. These results indicate that the fluorogenic nuclease RT-PCR assay should provide a potentially high-throughput, PCR-based method to detect rodent coronaviruses in infected rodents and contaminated biological materials.


title: Current and predicted trends in the production, consumption and trade of live animals and their products.

text: Changes in livestock production, driven by both demand- and supply-side factors, have been significant worldwide. Though historically the developed world was a large supplier of meat and livestock for the developing world, the developing world has rapidly increased production and is meeting more of its growing domestic demand. Many regions of the developing world, however, do not produce enough currently to meet their domestic demand and continue to import more than they produce. There are exceptions, such as Brazil, Thailand, the People's Republic of China and India, where growth in livestock production has been rapid. It is anticipated that in the future many of the developing countries will increase domestic production to meet growing domestic demand. By 2030, beef will probably still be the most significant meat import of developing countries and milk will have more than doubled as a net export of the developed world.


title: β-coronavirus infectious diseases: recommended strategies for the prevention and control of transmission.

text: In recent years, the incidence and mortality of infectious diseases in China are increasing. Infectious diseases, especially new infectious diseases, seriously threaten people's lives. Recent works found that most of the emerging infectious diseases come from wild life. At the same time, the impact of human activities on the environment has further deteriorated the living environment of wildlife. However, with the conducted in-depth research on virus, human beings increase the risk of getting infected. Taking Beta Coronavirus as the example, we analyzed the transmission risks of coronavirus in the prevention and control of the outbreaks of emerging infectious diseases, and recommend the prevention and control strategies before, during and after the viral outbreak. Additional works are urgently needed to better define the biological and epidemiological characteristics of these viruses.


title: [Immunological characteristics of 1291 cases of severe acute respiratory syndrome in Beijing].

text: OBJECTIVE To investigate the immunological characteristics of the cases of severe acute respiratory syndrome (SARS) in Beijing City. METHODS Clinical data of 1291 patients with SARS from March to July 2003 in Beijing City were retrospectively analyzed. RESULTS In patients with SARS, the absolute numbers of white blood cells, lymphocytes and CD(3), CD(4) and CD(8) lymphocyte subsets decreased during the early period of the disease, being manifested in 56.91%, 88.26%, 47.96%, 45.56% and 41.10% of the patients, respectively. During the first 3 days the median numbers of CD(3), CD(4) and CD(8) were 425 x 10(6)/L, 223 x 10(6)/L, 170 x 10(6)/L, respectively, being the lowest values in the course of the disease. During the second week the corresponding numbers were 536 x 10(6)/L, 267 x 10(6)/L, 224 x 10(6)/L, respectively; they returned to normal by the fourth week (P < 0.05), showing a trend of gradual increase during the disease progression. Comparison of different time points of the same cases also showed that CD(3), CD(4) and CD(8) were lowest in the first 1 - 3 days. The median number of CD(3) was higher (954 x 10(6)/L) during week 3, and there was no significant difference among other 3 weeks (P > 0.05). In the early period of the disease the CRP increased but ESR, C(3) and C(4) were still in normal ranges. CONCLUSIONS In the early period of SARS, the WBC, lymphocytes, CD(3), CD(4) and CD(8) lymphocyte subsets decreased remarkably, and they tended to increase as the disease progressed. Simultaneous decreases in CD(3), CD(4) and CD(8) during the first week is a characteristic immunological change, which may facilitate the early diagnosis of SARS.


title: Demystifying bioterrorism: misinformation and misperceptions.

text: The true threat of bioterrorism remains mysterious and elusive to the common citizen. It principally has become the dominion of a few "experts", many of whom have limited apparent expertise, who have failed to effectively communicate the risks and realities to society, and have instead created an air of uncertainty surrounding the topic. Unlike the great classic deceptions of modern life (e.g., "the check is in the mail"), the misinformation and misperceptions associated with bioterrorism can be dangerous and are not merely humorous. Indeed, it is possible to grasp the facts as well as fallacies associated with bioterrorism, and, as a result, demystify this nightmare scenario and prepare for the "unthinkable".


title: Ecchymoses as an adverse effect of fluoxetine treatment.

text: We report a case of a 28-year-old major depressive female patient who manifested ecchymoses following fluoxetine use. After substitution of sertraline, her depression resolved after 4 weeks and ecchymoses 1.5 months latter. This is an unexplored side-effect with unknown long-term consequences that warrants further study.


title: Foamy virus envelope glycoprotein is sufficient for particle budding and release.

text: Foamy viruses (FVs) are classified in the family Retroviridae, but recent data have shown that they are not conventional retroviruses. Notably, several characteristics of their particle replication strategies are more similar to those of hepatitis B virus (HBV) than those of typical retroviruses. Compared to conventional retroviruses, which require only Gag proteins for budding and release of virus-like particles (VLPs), both FV and HBV require Env proteins. In the case of HBV, Env (S protein) alone is sufficient to form subviral particles (SVPs). Because FVs also depend on Env for budding, we tested whether FV Env alone could produce SVPs. The Env proteins of FV and murine leukemia virus (MuLV) were both released into cell culture supernatants and migrated into isopycnic gradients; however, unlike MuLV Env, FV Env displayed characteristics of SVPs. FV Env particles were of greater density than those of MuLV (1.11 versus 1.07 g/ml, respectively), which strongly suggested that the released proteins of FV Env were particulate. When we examined FV SVPs by immunoelectron microscopy, we found particles that were consistent in morphology, size, and staining with gold beads, similar to FV VLPs and unlike the particle-like structures of MuLV Env, which were more consistent with vesicles produced from nonspecific membrane "blebbing." Taken together, our results demonstrated that FV Env alone is sufficient for particle budding. This finding is unique among retroviruses and further demonstrated the similarities between FV and HBV.


title: Outcomes of the adjustable gastric band in a publicly funded obesity program.

text: BACKGROUND Laparoscopic adjustable gastric banding (LAGB) is considered a safe and effective treatment for severe obesity and obesity-related comorbidities. We sought to examine the outcome of LAGB delivered through a Canadian publicly funded obesity program. METHODS We retrospectively analysed the cases of patients who underwent LAGB within a comprehensive, multidisciplinary, publically funded obesity program. RESULTS A total of 178 patients underwent LAGB. Mean percentage total body weight loss at 1, 2 and 3 years was 15.8%, 20.7% and 20.3%, respectively. The most common short-term complication was postoperative nausea (19%). The medium-term complications included band migration (5.6%) and port site complications, band leakage and incisional hernia at 1% each. The reoperation rate was 4.5%. The mean surgery duration was 56 minutes and the mean length of stay was 1.4 days. The average numbers of clinic visits and band adjustments were highest in the first year. The most common investigation for postoperative symptoms was fluoroscopy (86%). An outcome comparison between the 2 generations of the REALIZE gastric band was inconclusive, requiring further data collection. CONCLUSION Publicly funded LAGB results in effective weight loss and acceptable safety over the short term. Our patients may represent a distinct population that differs from that in the private system. Long-term data are necessary to determine the cost-effectiveness of this important surgical option for severe obesity.


title: Exogenous ACE2 expression allows refractory cell lines to support severe acute respiratory syndrome coronavirus replication.

text: Of 30 cell lines and primary cells examined, productive severe acute respiratory syndrome coronavirus (Urbani strain) (SARS-CoV) infection after low-multiplicity inoculation was detected in only six: three African green monkey kidney epithelial cell lines (Vero, Vero E6, and MA104), a human colon epithelial line (CaCo-2), a porcine kidney epithelial line [PK(15)], and mink lung epithelial cells (Mv 1 Lu). SARS-CoV produced a lytic infection in Vero, Vero E6, and MA104 cells, but there was no visible cytopathic effect in Caco-2, Mv 1 Lu, or PK(15) cells. Multistep growth kinetics were identical in Vero E6 and MA104 cells, with maximum titer reached 24 h postinoculation (hpi). Virus titer was maximal 96 hpi in CaCo-2 cells, and virus was continually produced from infected CaCo-2 cells for at least 6 weeks after infection. CaCo-2 was the only human cell type of 13 tested that supported efficient SARS-CoV replication. Expression of the SARS-CoV receptor, angiotensin-converting enzyme 2 (ACE2), resulted in SARS-CoV replication in all refractory cell lines examined. Titers achieved were variable and dependent upon the method of ACE2 expression.


title: [Preparation and characterization of the epitopes of SARS coronavirus spike protein].

text: AIM To prepare the recombinant epitopes of SARS-CoV spike protein and study their antigenic property to spike protein. METHODS The epitopes of SARS-CoV spike protein were analyzed by epitope prediction software. A novel gene, named Z, encoding 16 predicted epitopes of spike protein was designed and synthesized using chemical method. Z gene was cloned into vector pET-28a(+), expressed in E.coli BL21(DE3) and purified by Ni(2+) affinity method. Z protein was used as antigen to immunize the rabbit and anti-Z sera was collected. Then the antigenic property of Z protein to SARS-CoV spike protein was analyzed by dot-blot and ELISA assay. RESULTS Z gene was successfully designed and expressed in E.coli BL21(DE3). Dot blot analysis showed that SARS-CoV spike protein, which was expressed and purified from mammal cells, can be detected by anti-Z sera from rabbit. ELISA analysis indicated the SARS-CoV antigen prepared from SARS-CoV lysates can be detected by anti-Z sera. CONCLUSION The predicted epitopes of Z protein can induce the development of SARS-CoV spike protein antibody in rabbits, which provides a new protein for developing vaccine against SARS-CoV.


title: Bovine coronavirus spike glycoprotein: localization of an immunodominant region at the amino-terminal end of S2.

text: We have identified the binding site of monoclonal antibodies (MAbs) against the S2 subunit of the bovine coronavirus spike (S) glycoprotein. The location of this site was first investigated by using prokaryotic expression of DNA restriction fragments covering the entire S gene. The amino acid sequence containing the antibody binding site was shortened from 70 to 20 amino acids by digestion of plasmid DNA with exonuclease III, followed by sequencing of the smallest digestion product encoding an immunoreactive fusion protein. Finally we synthesized a set of nonapeptides covering the 20 amino acid sequence extending from the N-terminal residue of the S2 subunit (Ala 769 to Tyr 798). MAbs reacted mainly with six consecutive overlapping peptides with the sequence TTGYRFTNFEPFTV. Polyclonal antibodies from hyperimmunized or convalescent animals reacted only with the recombinant proteins identified by MAbs, and the hyperimmune serum bound to the same set of peptides. This suggests that this highly conserved linear antigenic determinant corresponds to an immunodominant region. This region resembles both in location and immunodominance the linear determinant defined on the infectious bronchitis virus S2 subunit. The presence of similar regions in the N-terminal region of the S2 subunit of other coronaviruses is discussed.


title: [Systematic review and meta-analysis on the integrative traditional Chinese and Western medicine in treating SARS].

text: OBJECTIVE To systematically evaluate the effects of Chinese herbal medicine in treating severe acute respiratory syndrome (SARS). METHODS Electronic and manual searches was conducted to identify randomised controlled trials (RCTs) for comparing the effects of integrative traditional Chinese and Western medicine versus the effects of Western medicine alone in treating SARS. Methodological quality of trials was assessed in terms of generation of allocation sequence, allocation concealment, blinding, and intention-to-treat. Exploring analysis was carried out on the non-randomized clinical controlled studies. Two authors collected data independently and evaluated quality and results. The data were quantified by Meta-analysis. RESULTS Eight RCTs (488 SARS patients) were included. The methodological quality was generally low. Results of Meta-analysis showed that the integrative therapy could significantly reduce the mortality (relative risk 0.32 [95% CI 0.12 - 0.91]) and the occurrence of fungal infection in patients who received hormone (glucocorticoid) therapy (0.35: 0.14 - 0.90), shorten the duration of fever (weighted mean difference -0.83 day; -1.30 - -0.35) and symptom relieving time (-1.23; -2.09 - -0.37), reduce the persistent time of chest radiograph abnormalities (- 2.61; -3.74 - -1.49). But it showed no significant difference on quality of life or glucocorticoid dosage. Conclusion The integrative therapy may have beneficial effects in lowering mortality, reducing complications, shortening the duration of symptoms and fever existence and lessening pulmonary inflammation in SARS patients. The evidence is insufficient to come to the affirmative conclusion due to the low methodological quality of the included trials.


title: Nursing students' knowledge and practice of infection control precautions: an educational intervention.

text: AIM This paper is a report of the effectiveness of a purpose-designed education program in improving undergraduate nursing students' understanding and practice of infection control precautions. BACKGROUND The severe acute respiratory syndrome outbreak in 2003 highlighted that healthcare workers were under-prepared for such an epidemic. While many in-service education sessions were arranged by institutions in response to the outbreak, preservice nursing education has overlooked preparation for handling such infectious disease epidemics. METHOD A quasi-experimental design was used and a 16-hour, purpose-designed infection control education programme was implemented for preservice nursing students in southern Taiwan. Self-administered questionnaires were distributed at three time points during the period September 2005 to April 2006 to examine the sustainability and effectiveness of the intervention. RESULTS A total of 175 preservice nursing students participated in the study. Following the education programme, students in the intervention group showed a statistically significant improvement across time in their knowledge of these precautions [F(2, 180) = 13.53, P < 0.001] and confidence in resolving infection-related issues [F(1.79, 168.95) = 3.24] when compared with those in the control group. CONCLUSION To improve nursing students' capacity in responding to infectious epidemics, an educational programme that integrates the theme of infection precautions, learning theory and teaching strategies is recommended for all nursing institutes.


title: Challenges and opportunities for new protein crystallization strategies in structure-based drug design.

text: Structure-based drug design (SBDD) has emerged as a valuable pharmaceutical lead discovery tool, showing potential for accelerating the discovery process,while reducing developmental costs and boosting potencies of the drug that is ultimately selected. SBDD is an iterative, rational, lead compound sculpting process that involves both the synthesis of new derivatives and the evaluation of their binding to the target structure either through computational docking or elucidation of the target structure as a complex with the lead compound. This method heavily relies on the production of high resolution(< 2 Å) 3D structures of the drug target, obtained through X-ray crystallographic analysis, in the presence or absence of the drug candidate.The lack of generalized methods for high quality crystal production is still a major bottleneck in the process of macromolecular crystallization. This review provides a brief introduction to SBDD and describes several macromolecular crystallization strategies, with an emphasis on advances and challenges facing researchers in the field today. Recent trends in the development of more universal macromolecular crystallization techniques, particularly nucleation-based techniques that are applicable to both soluble and integral membrane proteins, are also discussed.


title: Laparoscopic liver surgery: new frontiers.

text: Laparoscopic liver resection (LHR) has shown classical advantages of minimally invasive surgery over open counterpart. In spite of introduction in early 1990's only few centres worldwide adapted LHR to routine practice. It was due to considerable technical challenges and uncertainty about oncologic outcomes. Surgical instrumentation and accumulation of surgical experience has largely enabled to solve many technical considerations. Intraoperative navigation options have also been improved. Consequently indications have been drastically expanded nearly reaching criteria equal to open liver resection in expert centres. Recent studies have verified oncologic integrity of LHR. However, mastering of LHR is still a quite demanding task limiting expansion of this patient friendly technique. This emphasizes the necessity of systematic training for laparoscopic liver surgery. This article reviews the state of the art of laparoscopic liver surgery lightening burning issues of research and clinical practice.


title: Seroconversion of pigs in contact with dogs exposed to canine coronavirus.

text: In order to determine if canine coronavirus (CCV) could be transmitted to pigs, two dogs were inoculated orally with virulent CCV. After 24 h, the dogs were moved to an isolation room that contained three three-day-old pigs. A wire mesh fence, allowing close contact between the animals, separated the dogs from the pigs. The dogs and pigs were observed for 14 days for clinical signs of disease. Samples of blood were obtained from dogs and pigs immediately before the dogs were inoculated with virus and 14 and 28 days later. The dogs developed mild clinical signs of an infection, but the pigs remained normal throughout the observation period. The dogs shed CCV for eight days after exposure. All three pigs developed neutralizing antibodies against CCV and transmissible gastroenteritis virus by 14 days after they were exposed to the dogs.


title: Macaque model for severe acute respiratory syndrome.

text: Rhesus and cynomolgus macaques were challenged with 10(7) PFU of a clinical isolate of the severe acute respiratory syndrome (SARS) coronavirus. Some of the animals developed a mild self-limited respiratory infection very different from that observed in humans with SARS. The macaque model as it currently exists will have limited utility in the study of SARS and the evaluation of therapies.


title: [SARS epidemic trends in Beijing since late April 2003].

text: Since May 8th, Beijing's severe acute respiratory syndrome (SARS) epidemic is on a continuous decline. Indicating and verifying measures taken since the end of April to be effective. Three stages of SARS have been distinguished in Beijing since late April, they are described in this article as rapid growth stage beginning late April until early May, decline and stable stage throughout the first week of May, and rapid decline stage since May 9th until present. These three stages depict the effectiveness of interventive measures against SARS in Beijing since late April. Information transparency towards the public is an import part among all measures taken, enhancing public awareness and understanding of the current situation and government action as well as introduce public action. Public awareness is an important factor, forming active support for government interventions.


title: [Study on the damage of liver in patients with SARS].

text: OBJECTIVE To study the changes in liver function and histopathology, and investigate the underlying mechanism and clinical significance of damage of liver in patients with severe acute respiratory syndrome (SARS). METHODS According the clinical diagnostic standard of atypical pneumonia of Ministry of Health P. R. China, liver function was assessed in 110 SARS patients admitted from February 2003 to June 2003. Of them 8 SARS patients died, and the livers were pathologically examined, and their liver function parameters were compared with that of the 35 healthy controls. RESULTS Alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), total bilirubin (TBil) of patients with SARS were higher than those of controls, they were (91.61+/-50.53) U/L vs. (32.91+/-10.56) U/L, (78.68+/-33.32) U/L vs. (29.43+/-8.89) U/L, (429.95+/-188.94) U/L vs. (200.83+/-44.86) U/L, (11.67+/-4.26) micromol/L vs. (8.44+/-3.86) micromol/L, all P<0.001. Albumin (ALB) and pro-albumin (PAB) of patients with SARS were lower than those of controls, they were (34.40+/-5.13) g/L vs. (42.09+/-6.79) g/L, (0.20+/-0.06) g/L vs. (0.34+/-0.05) g/L, both P<0.001. Direct reaction bilirubin (DBil), total bile acid (TBA), gamma-glutamyltransferase (GGT) and alkaline phosphatase (ALP) showed no marked difference between SARS patients and controls, all P>0.05. Non-specific inflammation in the liver was observed in pathological examination in 4 cases. ALT, AST, GGT and LDH were always 4 to 9 times of normal. The ratio of abnormality of ALT, AST and PAB were more than 80.0 percent, the ratio of abnormality of ALB was 42.7 percent, and less than 30.0 percent for other indexes. The average of LDH, ALT, and AST of dead patients were higher than those of the survivors. Histopathology of liver was non-specific hepatitis. CONCLUSION The patients with SARS are prone to have mild non-specific hepatitis. It seldom causes the typical symptoms of hepatitis and it is easy to be ignored in clinic.


title: Identification of pulmonary Oct-4+ stem/progenitor cells and demonstration of their susceptibility to SARS coronavirus (SARS-CoV) infection in vitro.

text: In this study, we report a serum-free culture system for primary neonatal pulmonary cells that can support the growth of octamer-binding transcription factor 4+ (Oct-4+) epithelial colonies with a surrounding mesenchymal stroma. In addition to Oct-4, these cells also express other stem cell markers such as stage-specific embryonic antigen 1 (SSEA-1), stem cell antigen 1 (Sca-1), and Clara cell secretion protein (CCSP) but not c-Kit, CD34, and p63, indicating that they represent a subpopulation of Clara cells that have been implicated as lung stem/progenitor cells in lung injury models. These colony cells can be kept for weeks in primary cultures and undergo terminal differentiation to alveolar type-2- and type-1-like pneumocytes sequentially when removed from the stroma. In addition, we have demonstrated the presence of Oct-4+ long-term BrdU label-retaining cells at the bronchoalveolar junction of neonatal lung, providing a link between the Oct-4+ cells in vivo and in vitro and strengthening their identity as putative neonatal lung stem/progenitor cells. Lastly, these Oct-4+ epithelial colony cells, which also express angiotensin-converting enzyme 2, are the target cells for severe acute respiratory syndrome coronavirus infection in primary cultures and support active virus replication leading to their own destruction. These observations imply the possible involvement of lung stem/progenitor cells, in addition to pneumocytes, in severe acute respiratory syndrome coronavirus infection, accounting for the continued deterioration of lung tissues and apparent loss of capacity for lung repair.


title: Identification of new human coronaviruses.

text: To date, there are still a variety of human infections with unknown etiology. Identification of previously unrecognized viral agents in patient samples is of great medical interest but remains a major technical challenge. Acute respiratory tract infections are responsible for considerable morbidity and mortality in humans and animals. A variety of viruses, bacteria and fungi are associated with respiratory tract illness. Most of the respiratory viruses belong to the Paramyxoviridae, Orthomyxoviridae, Picornaviridae, Adenoviridae and Coronaviridae families. No pathogens can be detected in a relatively large proportion of patients with respiratory disease, partially owing to limitations of current diagnostic assays but also since some infections are caused by as yet unknown pathogens. This review will focus on human coronaviruses. In the mid 1960s, two human coronaviruses were identified that cause the common cold: human coronaviruses (HCoV)-229E and HCoV-OC43. The recent outbreak of severe acute respiratory syndrome-CoV and subsequent identification of two additional human coronaviruses (HCoV-NL63 and HCoV-HKU1) has drawn attention to this virus family. This review summarizes the knowledge of current methodologies for identifying novel human coronavirus species. Furthermore, information on the discovery of known human coronaviruses will be presented.


title: Prediction of postoperative alignment in patients undergoing anterior cervical fusion using autologous vertebral bone grafting.

text: Cervical anterior fusion is useful for the treatment of patients with cervical disease, but carries the risk of postoperative worsening of the cervical spine alignment and kyphosis. Preoperative prediction of the risk for kyphosis could help to avoid such postoperative complications. We attempted to predict the postoperative development of kyphosis in 59 patients scheduled for cervical anterior fusion using autologous vertebral bone grafting. Of these, 36 underwent single and 23 two level fusion with median follow up of 58.2 months. Whole spine alignment, fused segment alignment, and mobility of the intervertebral disc were compared using pre- and postoperative radiographs. Alignment of the whole spine changed from 13.7 degrees to 11.1 degrees , and the angle of the fused segment changed from 3.6 degrees to -3.2 degrees . Postoperative alignment was not worse than the preoperative flexion posture in any of the 59 patients. Forty-nine patients with preoperative lordotic alignment did not develop postoperative kyphosis. Nine patients with preoperative straight alignment had several types of postoperative alignment, including kyphosis. Two patients with postoperative kyphosis showed marked loss in the angle of the fused segment, which affected mobility to flexion of the fused segment. One patient with preoperative kyphotic alignment manifested postoperative kyphosis. The postoperative alignment was influenced by the preoperative alignment. Kyphosis may develop in patients with preoperative straight alignment and large mobility to flexion of the fused segment. This information is useful for surgical planning of anterior cervical fusion using autologous vertebral bone grafting.


RABBITS.


title: Internal Carotid Artery S-Shaped Curve as a Marker of Fibromuscular Dysplasia in Dissection-Related Acute Ischemic Stroke.

text: BACKGROUND AND PURPOSE Craniocervical fibromuscular dysplasia (FMD) is associated with dissections and with S-shaped curves in the internal carotid artery (ICA). We evaluated the occurrence of S-curves in patients presenting with acute strokes due to ICA steno-occlusive dissections. METHODS This was a retrospective review of the interventional databases of two academic tertiary-care institutions. The presence of ICA S-shaped curves, C-shaped curves, 360-degree loops, as well as classic FMD and atherosclerotic changes at the ICA bulb and curve/loop was determined. Cases of carotid dissections were compared with a control group (consecutive non-tandem anterior circulation strokes). RESULTS Twenty-four patients with carotid dissections were compared to 92 controls. Baseline characteristics and procedural variables were similar, with the exception of younger age, less frequent history of hypertension, diabetes, atrial fibrillation and stent retriever use in patients with dissections. The rates of mTICI2b-3 reperfusion, parenchymal hematoma, good outcome and mortality were similar amongst groups. The frequency of S-curves (any side without superimposed atherosclerosis) was 29% in the dissection group versus 7% in controls (p < 0.01). S-curves were typically mirror images within the dissection group (85% had bilateral occurrence). The frequency of C-shaped and 360-degree curves was similar between groups. FMD changes within the craniocervical arteries were statistically more common in dissection patients. Ten patients (41%) of the dissection group had S-curves or classic FMD changes. Multivariate analysis indicated that S-curves were independently associated with the presence of dissections. CONCLUSION S-shaped ICA curves are predictably bilateral, highly associated with carotid dissections in patients with moderate to severe strokes, and may suggest an underlying presence of FMD.


title: [Spontaneous coronavirus infection in monkeys].

text: A high level of the spread of coronavirus (CV) infection among hamadryas baboons and macaques of different species (about 50%), both resident in the animal house and imported, has been established. The tropism of CV to the gastrointestinal and respiratory tracts has been demonstrated. The course of spontaneous CV infection is accompanied by enterocolitis and/or pneumonia with periodic exacerbations, or takes the inapparent form. Cases of virus persistence have also been noted. Infected macaques exhibited an increase in the titers of antibodies to their own CV strain isolated from these animals, as well as to antigenically related human CV strain 0043. Spontaneous CV infection in monkeys may be used for solving some obscure problems of the pathogenesis and epidemiology of CV infection in humans.


title: Complement activation is required for induction of a protective antibody response against West Nile virus infection.

text: Infection with West Nile virus (WNV) causes a severe infection of the central nervous system (CNS) with higher levels of morbidity and mortality in the elderly and the immunocompromised. Experiments with mice have begun to define how the innate and adaptive immune responses function to limit infection. Here, we demonstrate that the complement system, a major component of innate immunity, controls WNV infection in vitro primarily in an antibody-dependent manner by neutralizing virus particles in solution and lysing WNV-infected cells. More decisively, mice that genetically lack the third component of complement or complement receptor 1 (CR1) and CR2 developed increased CNS virus burdens and were vulnerable to lethal infection at a low dose of WNV. Both C3-deficient and CR1- and CR2-deficient mice also had significant deficits in their humoral responses after infection with markedly reduced levels of specific anti-WNV immunoglobulin M (IgM) and IgG. Overall, these results suggest that complement controls WNV infection, in part through its ability to induce a protective antibody response.


title: [Progress on development and research of coronavirus based vector].

text: Due to the progress of targeted recombination and reverse genetics technique, it is possible to express foreign genes using coronaviruses as a vector via its unique transcription mechanism. Two types of coronavirus-based expression vectors have been developed; one is helper-dependent expression system and the other a single genome-based expression system. By modification of the infectious cDNA of the genome of coronaviruse, it has led to an efficient (> 50 microg/10(6)cells) and stable (> 30 passages) expression of the foreign gene. Moreover,several other features of coronavirus make it attractive as a vector. Firstly, the virulent coronaviruse can be converted into nonvirulent viruse by deleting the nonessential, group-specific genes. Secondly, the species and tissue tropism of coronaviruses can be manipulated by modification of their attachment and fusion protein S. Thus, coronavirus-based vector is becoming a promising and useful tool for vaccine development and gene delivery.


title: Blastomycosis in nondomestic felids.

text: Blastomycosis was diagnosed in six nondomestic felids from eastern Tennessee, including two Asian lions (Panthera leo persicus), one African lion (Panthera leo), one Siberian tiger (Panthera tigris), one cheetah (Acinonyx jubatus), and one snow leopard (Panthera uncia). Clinical signs included lethargy, anorexia, weight loss, dyspnea, sneezing. ataxia, and paresis. Variable nonspecific changes included leukocytosis, monocytosis, moderate left shift of neutrophils, moderate hypercalcemia, hyperproteinemia, and hyperglobulinemia. Thoracic radiographs revealed interstitial and alveolar changes, consolidation or collapse of a lung lobe, bullae formation, and a pulmonary mass. Agar gel immunodiffusion (AGID) serology for Blastomyces dermatitidis was performed in five felids and was positive in three. The tiger had cerebral blastomycosis and was positive for AGID serologic tests of both cerebrospinal fluid and serum. One percutaneous lung aspirate in the snow leopard and one bronchial aspirate in an Asian lion demonstrated B. dermatitidis organisms. whereas tracheal wash samples and a nasal discharge were nondiagnostic in others. Treatment with itraconazole was attempted in four cats. The tiger improved before euthanasia, whereas the others did not survive beyond initial treatments. In four felids, B. dermatitidis was found in the lungs and tracheobronchial lymph nodes associated with a florid pyogranulomatous reaction; the tiger had a pyogranulomatous encephalomyelitis, and the cheetah had a single pulmonary granuloma. Thoracic radiography, cytologic examination of lung lesion aspirates, and B. dermatitidis AGID serology should be performed on clinically ill zoo felids in endemic areas to rule out blastomycosis.


title: Evaluation of affymetrix severe acute respiratory syndrome resequencing GeneChips in characterization of the genomes of two strains of coronavirus infecting humans.

text: Severe acute respiratory syndrome (SARS) was discovered during a recent global outbreak of atypical pneumonia. A number of immunologic and molecular studies of the clinical samples led to the conclusion that a novel coronavirus (SARS-CoV) was associated with the outbreak. Later, a SARS resequencing GeneChip was developed by Affymetrix to characterize the complete genome of SARS-CoV on a single GeneChip. The present study was carried out to evaluate the performance of SARS resequencing GeneChips. Two human SARS-CoV strains (CDC#200301157 and Urbani) were resequenced by the SARS GeneChips. Five overlapping PCR amplicons were generated for each strain and hybridized with these GeneChips. The successfully hybridized GeneChips generated nucleotide sequences of nearly complete genomes for the two SARS-CoV strains with an average call rate of 94.6%. Multiple alignments of nucleotide sequences obtained from SARS GeneChips and conventional sequencing revealed full concordance. Furthermore, the GeneChip-based analysis revealed no additional polymorphic sites. The results of this study suggest that GeneChip-based genome characterization is fast and reproducible. Thus, SARS resequencing GeneChips may be employed as an alternate tool to obtain genome sequences of SARS-CoV strains pathogenic for humans in order to further understand the transmission dynamics of these viruses.


title: Infectious diseases notification practices, Victoria 2013.

text: INTRODUCTION Infectious disease notification practices in Victoria were reviewed to identify areas for potential improvement. METHODS Confirmed or probable cases of certain infectious diseases required to be notified to the Department of Health and Human Services (DHHS) Victoria in 2013, excluding elevated blood lead, foodborne or water-borne illness with 2 or more related cases and chlamydial infection, were analysed according to: notification source of doctor ± laboratory vs. laboratory-only; routine follow-up by public health staff for selected conditions vs. not routine; priority for Indigenous status reporting for 18 priority conditions with a target of ≥ 95% completeness vs. other conditions with a target of ≥ 80% completeness; and urgency of notification (conditions requiring immediate [same day] notification vs. conditions requiring notification within 5 days). RESULTS Almost half (49%) the 34,893 confirmed and probable cases were notified by laboratory report alone. Indigenous status was complete for 48% of cases. Indigenous status was more likely to be completed for conditions with active vs. no active follow-up (RR 1.88 (95% CI 1.84-1.92)) and priority conditions for Indigenous status reporting vs. other conditions (RR 1.62 (95% CI 1.59-1.66)). Among conditions without active follow-up, doctor-notified cases had more complete Indigenous status reporting than laboratory-only notified cases (86% vs. 6%, RR 15.06 (95% CI 14.15-16.03)). Fewer notifications requiring same day notification were received within the legislated time frame (59%) than notifications required to be notified within 5 days (90%). DISCUSSION DHHS Victoria handles a large volume of infectious disease notifications. Incomplete Indigenous status reporting, particularly for conditions without active follow-up, and delayed notification of conditions requiring immediate attention warrant attention. These findings will be used to improve notification practices in Victoria. Commun Dis Intell 2016;40(3):E317-E325.


title: [Epidemiological characteristics of 403 patients with severe acute respiratory syndrome in Haidian district, Beijing].

text: OBJECTIVE To analyze epidemiological features of severe acute respiratory syndrome (SARS) in Haidian district, Beijing. METHODS Each SARS case was interviewed by trained investigator using standardized questionnaire followed a descriptive analysis. RESULTS Four hundred and three SARS cases were identified and 27 of them died from March 18 and May 31, 2003. The incidence rate of SARS was 18.0/100,000 with case fatality rate as 6.7% in Haidian district, Beijing. Seventy-four percent of patients were adults with higher risk in age group of 20 - 29 year. SARS patients were scattered around in 32 out of 33 streets and villages in this district. The disease appeared to be sporadic but the case of outbreaks in family or university only seen in three streets. The course of SARS epidemic in this district could be divided into three phases: initial-which last for days, peak-21 days and then rapid decline-for 26 days. Number of patients having had a history of close contact to other SARS were gradually decreasing along with the process of the epidemics (trend chi(2) = 8.800, P = 0.003). Seventy-two point seven percent of the SARS cases had been exposed to the injection in the hospital settings. When the epidemics came to a rapid decline, 85.7% of the patients diagnosed during that period could be traced down to have had the history of contacting SARS cases within their own families. The distribution of occupation was also showed significantly different in the three respective stages (chi(2) = 36.41, P < 0.01). Among the patients who could not be identified as having confirmed contact history, 26.6% having had outward activities and 47.6% of them visited hospitals, especially during the peak stage. CONCLUSION The intensity of SARS epidemic among the residents of Haidian district was recognized as similar to the other parts of Beijing. Nosocomial infection in hospital settings was most important cause responsible for the transmission of SARS in this district.


title: Proteomic analysis of SARS associated coronavirus using two-dimensional liquid chromatography mass spectrometry and one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by mass spectroemtric analysis.

text: The proteomes of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) and its infected Vero E6 cells were detected in the present study. The cytosol and nucleus fractions of virus-infected cells as well as the crude virions were analyzed either by one-dimensional electrophoresis followed by ESI-MS/MS identification or by shotgun strategy with two-dimensional liquid chromatography-ESI-MS/MS. For the first time, all of the four predicted structural proteins of SARS-CoV were identified, including S (Spike), M (Membrane), N (Nucleocapsid), and E (Envolope) proteins. In addition, a novel phosphorylated site of M protein was observed. The combination of these gel-base and non-gel methods provides fast and complimentary approaches to SARS-CoV proteome and can be widely used in the analysis of other viruses.


Progressive encephalomyelopathy and cerebellar degeneration in a captive-bred snow leopard (Uncia uncia).


title: Unilateral laminotomy with bilateral decompression for lumbar spinal stenosis: short-term risks in elderly individuals.

text: The aim of this retrospective study was to compare the short-term (4 weeks) surgical complications after unilateral laminotomy for bilateral decompression (ULBD) in patients with lumbar spinal stenosis, below and above age 65. Twenty-nine consecutive patients operated upon in 2011 were included: 12 below age 65, and 17 above age 65. Five complications (17.1%) were observed, all in the older age group: 3 dural tears, one root injury, and one subcutaneous collection requiring re-operation. The difference between groups was not statistically significant (p = 0.0521). Moreover, comparable or higher complication rates were noted after alternative techniques. The CRP level, obtained one day postoperatively, was lower than after alternative procedures, pleading for the limited invasiveness of ULBD. Old age seems to be associated with slightly elevated rates of short-term surgical complications, but without an increment in morbidity. Unilateral laminotomy for bilateral decompression does not carry an excessive risk in the elderly population.