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

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


title: Lung damage may induce thrombocytopenia.

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


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

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


title: Single incision laparoscopic cholecystectomy.

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


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

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


docid: wuoypevi

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

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


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

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


Suicide prevention and intervention: a call to all doctors.


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


The avian coronavirus multiplication strategy.


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

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


title: Cardiovascular complications of severe acute respiratory syndrome.

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


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

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


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

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


docid: zjvbibcv

title: Paradoxical inflammation revisited: muraglitazar and cardiovascular risk.


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

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


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

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


title: Capnography in the pediatric emergency department: clinical applications

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


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

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


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

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


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

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


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

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


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

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


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

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


COVID-19: An Unintended Force for Medical Revolution?


Humanized mice develop coronavirus respiratory disease.


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


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

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


title: Laparoscopic repair of perineal hernia.

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


title: Effect of Pinus massoniana Lamb. bark extract on lytic cycle of Epstein-Barr virus.

text: Pinus massoniana bark extract (PMBE) at a concentration of 60 microg/mL or more inhibits the expression of Epstein-Barr virus (EBV) lytic proteins, such as Rta, Zta, and EA-D. EBV lytic cycle was blocked by inhibiting the transcription of immediate-early genes. The results suggest that the PMBE has anti-EBV activity. Thus, the extract is potentially useful in preventing the lytic development of EBV in vitro.


title: Effect of mutations in the mouse hepatitis virus 3'(+)42 protein binding element on RNA replication.

text: The mouse hepatitis virus (MHV) genome's 3' untranslated region contains cis-acting sequences necessary for replication. Studies of MHV and other coronaviruses have indicated a role for RNA secondary and tertiary elements in replication. Previous work in our laboratory has identified four proteins which form ribonucleoprotein complexes with the 3'-terminal 42 nucleotides [3'(+)42] of the MHV genome. Defective interfering (DI) RNA replication assays have demonstrated a role for the 3'(+)42 host protein binding element in the MHV life cycle. Using gel mobility shift RNase T1 protection assays and secondary structure modeling, we have characterized a possible role for RNA secondary structure in host protein binding to the 3'-terminal 42-nucleotide element. Additionally we have identified a role for the 3'-terminal 42-nucleotide host protein binding element in RNA replication and transcription using DI RNA replication assays and targeted recombination and by directly constructing mutants in this protein binding element using a recently described MHV reverse genetic system. DI RNA replication assays demonstrated that mutations in the 3'(+)42 host protein binding element had a deleterious effect on the accumulation of DI RNA. When the identical mutations were directly inserted into the MHV genome, most mutant genomes were viable but formed smaller plaques than the wild-type parent virus. One mutant was not viable. This mutant directed the synthesis of genome-sized negative-sense RNA approximately as efficiently as the wild type did but had a defect in subgenomic mRNA synthesis. These results point to a potential role for sequences at the extreme 3' end of the MHV genome in subgenomic RNA synthesis.


title: [In-hospital treatment of COVID-19 patients].

text: Covid-19 infection is now a pandemic disease. Most patients have mild symptoms without the need for hospitalization. On average, 5% of the COVID-19-positive patients have severe respiratory symptoms. We present 3 patients aged 45, 67 and 75 years, who were admitted to the ICU with severe respiratory symptoms. We describe the presentation of the COVID-19 patients on the emergency department, the development of the disease, and the treatment of these patients on the ICU.


title: Bovine virus diarrhoea virus: an introduction.

text: In view of the recently established genome organisation of pestiviruses, their classification as members of the togavirus family is no longer tenable. They should rather be provisionally considered as a new genus of the Flaviviridae, irrespective of differences in the nonstructural genes. Like other positive-stranded RNA viruses, pestiviruses are highly variable; apart from point mutations, recombinations are expected to contribute to their capricious behaviour. One trait of expected pathogenetic significance in infections with bovine virus diarrhoea virus is a change from the non-cytopathogenic to a cytopathogenic biotype. Cooperation of both variants in an animal to produce the severe disease picture known as mucosal disease is unique in virology; elucidation of this mechanism may shed light on the pathogenesis of other sporadic diseases with suspected viral origin.


docid: xxbtxtcx

title: Bats and human emerging diseases.


title: What have we learnt from SARS?

text: With outbreaks of infectious disease emerging from animal sources, we have learnt to expect the unexpected. We were, and are, expecting a new influenza A pandemic, but no one predicted the emergence of an unknown coronavirus (CoV) as a deadly human pathogen. Thanks to the preparedness of the international network of influenza researchers and laboratories, the cause of severe acute respiratory syndrome (SARS) was rapidly identified, but there is no complacency over the global or local management of the epidemic in terms of public health logistics. The human population was lucky that only a small proportion of infected persons proved to be highly infectious to others, and that they did not become so before they felt ill. These were the features that helped to make the outbreak containable. The next outbreak of another kind of transmissible disease may well be quite different.


title: Coronavirus JHM: intracellular protein synthesis.

text: Coronavirus JHM contained six major proteins, four of which were glycosylated. The proteins were gp170, gp98, gp65, p60, gp25 and p23. Sac(-) cells infected with JHM shut off host cell protein synthesis, and the synthesis of three major (150K, 60K and 23K) and three minor (65K, 30K and 14K) polypeptides was detected by pulse-labelling with 35S-methionine. Antiserum directed against purified virus proteins specifically immunoprecipitated the three major intracellular species and also the 65K minor species. During a chase period, species 150K and 23K were processed and three new immunoprecipitable species, 170K, 98K and 25K appeared. The intracellular species 170K, 98K, 65K, 60K, 25K and 23K co-electrophoresed with virion proteins. Two-dimensional gel electrophoresis of infected cell polypeptides showed that the 60K, 23K, 25K and 14K species were relatively basic polypeptides whilst the 98K and 170K were relatively acidic and heterogeneously charged polypeptides. Additionally, a charge-size modification of the 23K species during processing was detected, which was not apparent using one-dimensional gel analysis.


docid: roevjttz

title: Gold standard for humidification: heat and moisture exchangers, heated humidifiers, or both?


Pathogenesis of SARS coronavirus infection using human lung epithelial cells: an in vitro model.


title: Investigation of an imported case of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) infection in Florence, Italy, May to June 2013.

text: On 31 May 2013, the first case of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) infection in Italy was laboratory confirmed in a previously healthy adult man, who developed pneumonia with moderate respiratory distress after returning from a holiday in Jordan. Two secondary cases were identified through contact tracing, among family members and colleagues who had not previously travelled abroad. Both secondary cases developed mild illness. All three patients recovered fully.


title: [Acute respiratory disease caused by coronaviruses].

text: Serological evidence of coronavirus aetiology was found in 11 patients out of 218 persons examined (5%) in the period of 1986-1987, and in 23 cases out of 311 patients (7.4%) in the period of 1987-1988. Antibody titres with a minimum of four-time elevated values in paired sera using the ELISA method were considered conclusive. Coronaviruses 229E and OC43 were employed. During the first period, more persons were infected by coronavirus 229E, while in the second period, by coronavirus OC43. The disease was equally spread over all the months of the years. For example, in the 1987-1988 period, most infections occurred in February, when out of 60 examined patients, 9 were coronavirus positive (15%).


title: Law and public health: beyond emergency preparedness.

text: This Article examines three questions: What is public health? What is public health law? What roles can lawyers play in public health? It first describes the breadth of public health, highlighting six trends shaping its future: social determinants of health; synergy between medicine and public health; shifts in focus from external (e.g., environmental and social) to internal (behavioral) risks to health; federalization of public health law; globalization of health risks and responses; and bioterrorism. Because the domains of law that apply to public health are equally broad, the Article next offers a conceptual framework for identifying the types of laws most suitable to different public health problems. Finally, the role of lawyers in the applied field of public health law is examined, first to encourage attention to law's effect on health, even laws having little apparent relationship to health; and second, to recognize that laws intended to achieve specific health outcomes may affect broader legal principles. Lawyers have a unique role to play in ensuring that the legal principles used to promote health also preserve justice.


title: [Diagnostic imaging findings in COVID-19].

text: This is a review of diagnostic imaging findings in coronavirus disease-2019 (COVID-19). CT findings are frequently bilateral, multilobar and peripheral ground-glass opacities with vascular enlargements. Consolidations often appear during progression as well as crazy paving and reticulation. Lymphadenopathy, pleural effusions and pneumothorax are rare and should raise concern for other disease. Initially the CT can be normal, and the diagnostic performance for COVID-19 is not optimal. However, findings of characteristic ground-glass opacities during the ongoing epidemic should lead to considerations, that the patient may have COVID-19.


title: Enhanced growth of a murine coronavirus in transformed mouse cells.

text: Plaque formation by A59 virus, a murine coronavirus, was facilitated in AL/N and Balb mouse cells transformed by polyoma virus, simian virus 40, murine sarcoma virus, or mammary tumor virus. In these virus-transformed cells, A59 virus plaques were larger, they appeared earlier, and plaquing efficiencies were higher than in normal, untransformed cells. "Spontaneously" transformed AL/N cells behaved similarly to untransformed cells, whereas "spontaneously" transformed Balb cells resembled virus-transformed cell hosts. Both untransformed and transformed AL/N and Balb cells were permissive hosts for A59 virus. However, multiplication of A59 virus was enhanced at least fivefold in the virus-transformed AL/N cell hosts. Larger differences (100-fold or greater) in A59 virus production were obtained during the first cycle of infection in Balb cells at low multiplicities and in AL/N cells after multiple cycles of virus growth. In virus-transformed and "spontaneously" transformed Balb cells, A59 virus induced extensive syncytia formation.


docid: hqkuekzs

title: [Expression of recombinant spike protein of SARS-coronavirus in vaccinia virus and analysis of its immunogenicity].

text: A recombinant vaccinia virus (rWR-SARS-S)expressing spike protein of severe acute respiratory syndrome coronavirus was constructed. The expression of full length recombinant SARS spike protein (rSS) in HeLa cells possessing specific reaction ability to chicken anti-sera was confirmed by SDS-PAGE and Western-blot (190 kD). HeLa cells infected with rWR-SARS-S also showed high sensitivity in detecting specific serum antibody by indirect immunofluoresence assay (IFA). The results above indicated that the availability of such a faithful model system offers particular advantages for the study of SARS in that it reduces the need for direct manipulation of an exotic pathogen. In the absence of infectious SARS, we may safely carry out detailed biochemical and genetic manipulations to investigate features of viral replication and gene function, as well as explore new avenue for vaccine development.


docid: wzvdaygp

title: Increased risk of incident HIV during pregnancy in Rakai, Uganda: a prospective study.

text: BACKGROUND HIV acquisition is significantly higher during pregnancy than in the postpartum period. We did a prospective study to estimate HIV incidence rates during pregnancy and lactation. METHODS We assessed 2188 HIV-negative sexually active women with 2625 exposure intervals during pregnancy and 2887 intervals during breastfeeding, and 8473 non-pregnant and non-lactating women with 24,258 exposure intervals. Outcomes were HIV incidence rates per 100 person years and incidence rate ratios estimated by Poisson multivariate regression, with the non-pregnant or non-lactating women as the reference group. We also assessed the husbands of the married women to study male risk behaviours. FINDINGS HIV incidence rates were 2.3 per 100 person years during pregnancy, 1.3 per 100 person years during breastfeeding, and 1.1 per 100 person years in the non-pregnant and non-lactating women. The adjusted incidence rate ratios were 2.16 (95% CI 1.39-3.37) during pregnancy and 1.16 (0.82-1.63) during breastfeeding. Pregnant women and their male partners reported significantly fewer external sexual partners than did the other groups. In married pregnant women who had a sexual relationship with their male spouses, the HIV incidence rate ratio was 1.36 (0.63-2.93). In married pregnant women in HIV-discordant relationships (ie, with HIV-positive men) the incidence rate ratio was 1.76 (0.62-4.03). INTERPRETATION The risk of HIV acquisition rises during pregnancy. This change is unlikely to be due to sexual risk behaviours, but might be attributable to hormonal changes affecting the genital tract mucosa or immune responses. HIV prevention efforts are needed during pregnancy to protect mothers and their infants.


title: Glial response in the central nervous system of cats with feline infectious peritonitis.

text: OBJECTIVES The aim of the study was to evaluate central nervous system (CNS) lesions in non-effusive and effusive cases of feline infectious peritonitis (FIP) regarding aspects related to astrocytic and microglial reactions. METHODS Five necropsied cats that were naturally infected with FIP virus, confirmed by reverse transcriptase polymerase chain reaction and immunohistochemistry, with different intensities of CNS lesions, were studied. Brain and cerebellum were evaluated by light microscopy and immunohistochemistry for glial fibrillary acidic protein (GFAP) and vimentin to assess astrocytic morphology, and lectin histochemistry for Ricinus communis agglutinin-I (RCA-I) to detect microglia was performed to evaluate the glial response in the CNS of cats with FIP. RESULTS An important astrocytic response in many areas of the CNS of all cats, including the periventricular areas of lateral ventricles and fourth ventricle, the molecular layer of the cerebellum and cerebral cortex, was visualized. This astrocytic reactivity was associated with areas of granulomatous or pyogranulomatous vasculitis/perivasculitis in most cases, and it was characterized by multifocal to coalescing astrocytosis and astrogliosis with an increase in the expression of intermediate filaments, such as GFAP. However, astrocytes exhibited strong vimentin expression in neuroparenchyma with severe inflammatory and necrotic changes, but GFAP expression was mild or absent in these cases. A microglial response was present only in severe lesions, and RCA-I expression was detected primarily in gitter cells and resting microglia. CONCLUSIONS AND RELEVANCE The present study indicates a strong astrocytic response, including the presence of many less differentiated vimentin-positive astrocytes and gitter cells positive for RCA-1 in severe lesions in the CNS of cats with FIP.


title: Depression in patients with euthyroid chronic autoimmune thyroiditis.

text: Depression and chronic autoimmune thyroiditis are common diseases. The aim of this study was to determine whether the frequency of depression is elevated in patients with chronic autoimmune thyroiditis and normal thyroid function. A total of 201 subjects were included, of whom 107 and 94 participants were healthy or had euthyroid chronic autoimmune thyroiditis, respectively. Serum thyroid hormone levels and thyroid auto-antibodies were measured in all subjects. All participants were evaluated with Hamilton Depression Rating Scale (HDRS). HDRS scores of patients with euthyroid chronic autoimmune thyroiditis were higher than in healthy participants. This study suggests a possible association between depression and chronic autoimmune thyroiditis. The depression risk may be increased in patients with chronic autoimmune thyroiditis and normal thyroid function.


title: [A preliminary report of SARS coronavirus specific RNA in SARS convalescents and medical person].

text: OBJECTIVE To observe the method of fluorescent-polymerase chain reaction (F-PCR) and gene-hip technique in detecting coronavirus in severe acute respiratory syndrome (SARS) and its value for clinical application. METHODS Serum of 60 SARS patients, 20 samples of serum and 20 samples of gargling fluid of medical staff of the "Fever Clinic", and one cDNA specimen obtained from one SARS suspect patient were examined with F-PCR diagnosis kit and gene-chip technique for SARS coronavirus. RESULTS Three methods, including DR Chip and two F-PCR kit from Zhongshan College Da'an gene stock company and Shanghai Fortune industrial joint-stock Co, Ltd, were used, and the results were all negative for all specimens, except one cDNA specimen, which was obtained from one SARS suspect patient, virus could be amplified by F-PCR. CONCLUSION Special RNA fragment of SARS virus has not been detected in gargling fluid and serum of SARS convalescents and medical staff.


title: Identification and epidemiology of severe respiratory disease due to novel swine-origin influenza A (H1N1) virus infection in Alberta.

text: BACKGROUND In March 2009, global surveillance started detecting cases of influenza-like illness in Mexico. By mid-April 2009, two pediatric patients were identified in the United States who were confirmed to be infected by a novel influenza A (H1N1) strain. The present article describes the first identified severe respiratory infection and the first death associated with pandemic H1N1 (pH1N1) in Canada. METHODS Enhanced public health and laboratory surveillance for pH1N1 was implemented throughout Alberta on April 24, 2009. Respiratory specimens from all patients with a respiratory illness and travel history or those presenting with a severe respiratory infection requiring hospitalization underwent screening for respiratory viruses using molecular methods. For the first severe case identified and the first death due to pH1N1, histocompatibility leukocyte antigens were compared by molecular methods. RESULTS The first death (a 39-year-old woman) occurred on April 28, 2009, and on May 1, 2009, a 10-year-old child presented with severe respiratory distress due to pH1N1. Both patients had no travel or contact with anyone who had travelled to Mexico; the cases were not linked. Histocompatibility antigen comparison of both patients did not identify any notable similarity. pH1N1 strains identified in Alberta did not differ from the Mexican strain. CONCLUSION Rapid transmission of pH1N1 continued to occur in Alberta following the first death and the first severe respiratory infection in Canada, which were identified without any apparent connection to Mexico or the United States. Contact tracing follow-up suggested that oseltamivir may have prevented ongoing transmission of pH1N1.


title: [Effects of underlying cerebrocardiovascular diseases on the incidence of critical conditions and multiple organs dysfunction syndrome in severe acute respiratory syndrome cases].

text: OBJECTIVE To investigate the effects of underlying cerebrocardiovascular diseases on the incidence of critical conditions and multiple organs dysfunction syndrome (MODS) among the severe acute respiratory syndrome (SARS) cases. METHODS A database of all SARS cases in Beijing in 2003 was established and the data of 1291 cases whose data were complete among them was analyzed. The effects of cerebrocardiovascular diseases, other comorbid diseases and absence of underlying diseases on the incidence of critical conditions and MODS in SARS cases were compared with chi-square test and logistic regression analysis. RESULTS The incidence rates of critical conditions and MODS in SARS cases were 34.86% and 13.71% respectively. The incidence rates of critical conditions and MODS among the SARS cases without underlying diseases, with other comorbid diseases, and with cerebrocardiovascular diseases were 28.10%, 43.70%, and 58.18%, and 9.83%, 18.52%, and 27.27% respectively. The differences among the three groups were all statistically significant (all P < 0.0001). Logistic regression analysis showed that presence of cerebrocardiovascular diseases in SARS patients was independently associated with the risks of critical conditions and MODS in comparison with those without underlying diseases after adjusting for age and occupation. [odds ratio (OR) = 1.819, 95% CI = 1.276 approximately 2.592, and OR = 1.974, 95% CI = 1.273 approximately 3.060 respectively]. CONCLUSION The SARS cases with cerebrocardiovascular diseases have the highest incidence rate of critical conditions and MODS. Presence of Cerebrocardiovascular diseases is an important risk factor for critical conditions and MODS in the SARS cases.


Factors affecting isolation and propagation of bovine coronavirus in human rectal tumor-18 cell line.


title: Development of a competitive enzyme-linked immunosorbent assay for detection of turkey coronavirus antibodies.

text: A competitive enzyme-linked immunosorbent assay (cELISA) was developed for detection of turkey coronavirus (TCV) antibodies. The cELISA utilized a recombinant baculovirus (Autographa californica nuclear polyhedrosis virus)-expressed TCV nucleocapsid (N) protein and biotin-labeled TCV N protein-specific monoclonal antibody. Sensitivity and specificity of the cELISA for detection of TCV antibodies were determined by comparison with the indirect fluorescent antibody test (IFAT) with 1269 reference, experimentally derived, and field-origin sera. Sera with discordant cELISA and IFAT results were further evaluated by western immunoblot analyses. The cELISA detected antibodies specific for TCV and infectious bronchitis virus, a closely related coronavirus, but did not detect antibodies specific for other avian viruses. A high degree of concordance was observed between the cELISA and IFAT; sensitivity and specificity of the cELISA relative to IFAT were 92.9% and 96.2%, respectively. Western immunoblot analyses provided additional evidence of cELISA specificity. The findings indicate that the cELISA is a rapid, sensitive, and specific serologic test for detection of TCV antibodies in turkeys.


title: Pneumocystis carinii in a patient with pulmonary sarcoidosis and idiopathic CD4+ T lymphocytopenia.

text: A case of pulmonary sarcoidosis and idiopathic CD4+ T lymphocytopenia is reported. Pneumocystis carinii was detected in the bronchoalveolar lavage fluid of a young homosexual man who was asymptomatic without any evidence of congenital or acquired immunodeficiency but with a low CD4+ cell count. A clinical and histological diagnosis of pulmonary sarcoidosis was made. During follow up the patient had oral candidiasis and a CD4+ cell count persistently below 300/microliters. This case is highly suggestive of concurrent pulmonary sarcoidosis and idiopathic CD4+ T lymphocytopenia.


title: A structural model of picornavirus leader proteinases based on papain and bleomycin hydrolase.

text: The leader (L) proteinases of aphthoviruses (foot-and-mouth disease viruses) and equine rhinovirus serotypes 1 and 2 cleave themselves from the growing polyprotein. This cleavage occurs intramolecularly between the C terminus of the L proteinases and the N terminus of the subsequent protein VP4. The foot-and-mouth disease virus enzyme has been shown, in addition, to cleave at least one cellular protein, the eukaryotic initiation factor 4G. Mechanistically, inhibitor studies and sequence analysis have been used to classify the L proteinases as papain-like cysteine proteinases. However, sequence identity within the L proteinases themselves is low (between 18% and 32%) and only 14% between the L proteinases and papain. Secondary structure predictions, sequence alignments that take into account the positions of the essential catalytic residues, and structural considerations have been used in this study to investigate more closely the relationships between the L proteinases and papain. In spite of the low sequence identities, the analyses strongly suggest that the L proteinases of foot-and-mouth disease virus and of equine rhinovirus 1 have a similar overall fold to that of papain. Regions in the L proteinases corresponding to all five alpha-helices and seven beta-sheets of papain could be identified. Further comparisons with the proteinase bleomycin hydrolase, which also displays a papain topology in spite of important differences in size and amino acid sequence, support these conclusions and suggest how a C-terminal extension, present in all three L proteinases, and predicted to be an alpha-helix, might enable C-terminal self-processing to occur.


title: Epidemiological study of autism spectrum disorder in China.

text: The object of this study was to investigate the epidemiologic pattern of autism spectrum disorder in Chinese children. An autism spectrum disorder registry has been established in Hong Kong since 1986 by collecting data in a single center (the only university-affiliated child assessment center in Hong Kong). Since 1997, inpatient data from all public hospitals under the Hospital Authority have been stored in a central computerized program and retrieved from the Clinical Data Analysis and Reporting System. Clinical data have also been retrieved through the Clinical Data Analysis and Reporting System to ensure the completion of the registry, and these suspected cases have undergone the same diagnostic evaluation for autism spectrum disorder, as some of the new autism cases might be hospitalized in the public hospital. The incidence and prevalence of autism spectrum disorder have been calculated for the period of 1986 to 2005 using the population statistics available in the government for children less than 15 years old in Hong Kong. This study has included 4 247 206 person-years from 1986 to 2005 for children less than 15 years old and 1 174 322 person-years for those less than 5 years old in Hong Kong. Altogether, 645 children 0 to 4 years old with diagnoses of autism spectrum disorder were identified from 1986 to 2005. The estimated incidence of autism spectrum disorder was 5.49 per 10 000. The prevalence was 16.1 per 10 000 for children less than 15 years old for the same period. The male to female ratio was 6.58:1. This is the first reported epidemiologic study of autism spectrum disorder in Chinese children. The incidence rate is similar to those reported in Australia and North America and is lower than Europeans.


title: [Preliminary note on the circulation of some human respiratory viruses by wild birds].

text: With a view to detecting infections and carriage of human respiratory viruses in wild birds, 349 serum samples collected from 21 bird species -- sedentary ones and birds with large or limited migration areas -- were investigated. The following antigens were used: influenza virus A/Hong Kong 1/68 (H3N2)), A2 England 42/73 (H3N2), A2 Victoria 3/75 (H3N2), A/New Jersey 8/76 (HswN1), B/Hong Kong 5/72; parainfluenza virus type I Sendai and type II, coronavirus OC/43. An elevated incidence of antibodies to A2 Victoria 3/75 (H3N2) and parainfluenza type I virus was detected in herons (Ardea cinerea, Nyctycorax myctycorax). The high incidence of antibodies to B/Hong Kong 5/72 (30.7% of the samples exhibited significant titers) found in the crow (Corvus corone sardonius) is ascribed to the fact that this bird is carnivorus, feeding on corpses of mammals.


title: Detection of a bovine group C rotavirus from adult cows with diarrhea and reduced milk production.

text: Only two strains (Shintoku and porcine-like WD534tc) of group C rotavirus (GCR) from cattle have been reported to date. A GCR designated the Yamagata strain was the only pathogen detected in an outbreak of adult cow diarrhea accompanied by a decrease in milk production. The nucleotide sequences of the VP6 and VP7 genes from strain Yamagata were determined. Comparative sequence analysis showed that the sequence identities between strains Yamagata and Shintoku were markedly high in both VP6 gene (98.1%) and VP7 gene (93.5%), and that these strains belonged to the same clusters which were distinguished from GCRs from different host species in phylogenetic trees of these genes. These results suggested strongly that cattle species is one of the natural hosts of GCR infection, and that GCRs are a cause of adult cow diarrhea.


title: Expression and secretion of the bovine coronavirus hemagglutinin-esterase glycoprotein by insect cells infected with recombinant baculoviruses.

text: A cDNA fragment representing the hemagglutinin-esterase (HE) gene of bovine coronavirus (BCV) was inserted into the genome of Autographa californica nuclear polyhedrosis virus. Infection of insect cells with the recombinant virus resulted in the production of a 120-kilodalton disulfide-linked dimeric form of the BCV HE polypeptide. Deletion of the carboxy-terminal hydrophobic domain from the HE polypeptide resulted in secretion of a dimeric form of the truncated HE polypeptide. The acetylesterase activity of the BCV HE was detectable in insect cells expressing the BCV hemagglutinin and was inhibited by two monoclonal antibodies which also inhibit hemagglutination.


title: Comparison of Cross-linked and Non-Cross-linked Acellular Porcine Dermal Scaffolds for Long-term Full-Thickness Hernia Repair in a Small Animal Model.

text: BACKGROUND This study compared the strength of incorporation and biocompatibility of 2 porcine-derived grafts (cross-linked and non-cross-linked) in a rat hernia model. METHODS A standardized 2 × 4 cm(2) fascial defect was created in 30 Wistar rats and repaired with either a cross-linked or a non-cross-linked graft. The rats were killed 3, 6, and 12 months later. The strength of incorporation, vascularization, cellular invasion, foreign body reaction, and capsule formation were evaluated. RESULTS Both graft materials showed cellular ingrowth and neovascularization by 3 months postimplantation. The average level of cellularization was significantly higher in the non-cross-linked grafts than in the cross-linked grafts at 6 months (2 vs 1; P = .029). Vascularization was significantly higher in the non-cross-linked grafts than in the cross-linked grafts at 6 months postimplantation (2 vs 1; P = .029) and insignificant at 3 months (2 vs 1.75; P = .311) and 12 months (1 vs 0.67; P = 1). The maximum load and breaking strength of both biomaterials increased during the study period. Overall, the strength of incorporation of the non-cross-linked grafts increased from 3 months (0.75 MPa) to 12 months (3.06 MPa) postimplantation. The strength of incorporation of the cross-linked grafts also increased from 3 months (0.59 MPa) to 12 months (1.58 MPa) postimplantation. CONCLUSIONS The results of our study suggest that non-cross-linked grafts may be slightly more biocompatible and allow a more rapid and higher degree of cellular penetration and vascularization, resulting in stronger attachment to the tissues.


docid: wbuszumk

title: Effect of temperature on the detection of porcine epidemic diarrhea virus and transmissible gastroenteritis virus in fecal samples by reverse transcription-polymerase chain reaction.

text: The effect of storage temperature was determined for the detection of porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) in fecal samples from experimentally and naturally infected pigs by multiplex reverse transcription-polymerase chain reaction (RT-PCR). To examine the effect of storage temperature on the ability to detect PEDV and TGEV RNA by multiplex RT-PCR, fecal samples were stored for different temperatures (4, 21, 36, and 45 C) before extracting viral RNA. The PEDV and TGEV nucleic acids in fecal samples were stable up to 3 days at 4 C. The PEDV and TGEV nucleic acids were consistently detected in fecal samples up to 60 hours at 21 C and 24 hours only at 36 and 45 C. Thereafter, the number of positive signals declined. Positive signals were detected in fecal samples stored at 4 C by 240 hours. The PEDV and TGEV nucleic acids were consistently detected in fecal samples up to 60 hours at 21 C and 24 hours only at 36 and 45 C. The results of this study suggested that storage temperature has significant effect on the detection of PEDV and TGEV nucleic acids from fecal samples by multiplex RT-PCR.


title: Infection of primary cultures of human neural cells by human coronaviruses 229E and OC43.

text: We evaluated the ability of human coronaviruses to infect primary cultures of human neural cells. Double immunofluorescence with antibodies to virus and cell markers showed infection of fetal astrocytes and of adult microglia and astrocytes by strain OC43. RNA amplification revealed infection of fetal astrocytes, adult microglia, and a mixed culture of adult oligodendrocytes and astrocytes by strain 229E. Infectious virus was released only from fetal astrocytes, with higher titers for OC43. Human coronaviruses have the capacity to infect some cells of the central nervous system, although infection of adult cells appears abortive.


title: Cellular and humoral immunity following Snow Mountain virus challenge.

text: Little is known about the immune response to noroviruses. To elucidate the immunobiology of norovirus infection in humans, 15 volunteers were challenged with Snow Mountain virus (SMV), a genogroup 2 norovirus. We assessed the cellular and humoral immune response and infection by analyzing stool, serum, saliva, and peripheral blood mononuclear cell (PBMC) responses pre- and postchallenge. In contrast to Norwalk virus (NV), SMV infection was not dependent upon blood group secretor status. Nine of 15 volunteers were infected and showed a >/=4-fold increase over the prechallenge anti-SMV serum immunoglobulin G (IgG) titer, mostly subclass IgG1. Although serum IgG elicited by SMV infection was cross-reactive with Hawaii virus (HV), another genogroup 2 norovirus, salivary IgA was less cross-reactive. Neither SMV-elicited serum IgG nor salivary IgA cross-reacted with NV, a genogroup 1 norovirus. Significant increases in serum gamma interferon (IFN-gamma) and IL-2, but not IL-6 or IL-10, were noted on day 2 postchallenge. For the majority of volunteers, both infected and uninfected, PBMCs stimulated with norovirus virus-like particles secreted IFN-gamma and other Th1 cytokines, suggesting previous norovirus exposure in most volunteers. Like the IgG antibodies, the SMV-activated T cells were cross-reactive with HV but not NV. IFN-gamma production was dependent upon CD4(+) cells, consistent with a predominant, but not exclusive, Th1 response. To our knowledge, this is the first report characterizing T-cell and cytokine responses following live norovirus challenge.


title: Biosynthesis, purification, and characterization of the human coronavirus 229E 3C-like proteinase.

text: Coronavirus gene expression involves proteolytic processing of the gene 1-encoded polyprotein(s), and a key enzyme in this process is the viral 3C-like proteinase. In this report, we describe the biosynthesis of the human coronavirus 229E 3C-like proteinase in Escherichia coli and the enzymatic properties, inhibitor profile, and substrate specificity of the purified protein. Furthermore, we have introduced single amino acid substitutions and carboxyl-terminal deletions into the recombinant protein and determined the ability of these mutant 3C-like proteinases to catalyze the cleavage of a peptide substrate. Using this approach, we have identified the residues Cys-3109 and His-3006 as being indispensable for catalytic activity. Our results also support the involvement of His-3127 in substrate recognition, and they confirm the requirement of the carboxyl-terminal extension found in coronavirus 3C-like proteinases for enzymatic activity. These data provide experimental evidence for the relationship of coronavirus 3C-like proteinases to other viral chymotrypsin-like enzymes, but they also show that the coronavirus proteinase has additional, unique properties.


title: Quaternary structure of the severe acute respiratory syndrome (SARS) coronavirus main protease.

text: SARS (severe acute respiratory syndrome) has been one of the most severe viral infectious diseases last year and still remains as a highly risky public health problem around the world. Exploring the types of interactions responsible for structural stabilities of its component protein molecules constitutes one of the approaches to find a destabilization method for the virion particle. In this study, we performed a series of experiments to characterize the quaternary structure of the dimeric coronavirus main protease (M(pro), 3CL(pro)). By using the analytical ultracentrifuge, we demonstrated that the dimeric SARS coronavirus main protease exists as the major form in solution at protein concentration as low as 0.10 mg/mL at neutral pH. The enzyme started to dissociate at acidic and alkali pH values. Ionic strength has profound effect on the dimer stability indicating that the major force involved in the subunit association is ionic interactions. The effect of ionic strength on the protease molecule was reflected by the drastic change of electrostatic potential contour of the enzyme in the presence of NaCl. Analysis of the crystal structures indicated that the interfacial ionic interaction was attributed to the Arg-4...Glu-290 ion pair between the subunits. Detailed examination of the dimer-monomer equilibrium at different pH values reveals apparent pK(a) values of 8.0 +/- 0.2 and 5.0 +/- 0.1 for the Arg-4 and Glu-290, respectively. Mutation at these two positions reduces the association affinity between subunits, and the Glu-290 mutants had diminished enzyme activity. This information is useful in searching for substances that can intervene in the subunit association, which is attractive as a target to neutralize the virulence of SARS coronavirus.


title: Putting proteomics on target: activity-based profiling of ubiquitin and ubiquitin-like processing enzymes.

text: Modification of proteins with ubiquitin (Ub) and Ub-like modifiers (Ubls) plays a fundamental role in cell biology. As a consequence, proteomics-based efforts were developed to characterize proteins that are modified by Ub or Ubls. A more focused functional proteomics strategy relies on active-site probes based on the Ub/Ubl scaffold, which specifically targets Ub/Ubl-processing enzymes. Activity-based profiling with such tools led to the identification of novel gene products with Ub/Ubl-processing activity and uncovered novel control mechanisms regulating their activity. This review discusses recent advances in chemistry-based functional proteomics applications, and how this information can provide a framework for drug development against Ub/Ubl-processing enzymes.


title: Binding properties and anti-bacterial activities of V-region identical, human IgG and IgM antibodies, against group B Neisseria meningitidis.

text: We have constructed chimaeric (ch) mouse/human antibodies with identical binding regions isolated from the V-genes of two mouse parent hybridoma cell lines, with specificity against the P1.7 and P1.16 epitopes on the outer-membrane protein PorA on meningococci. The chimaeric antibodies can be used to analyse relationships between specificity, binding activity (avidity and kinetics), isotype (antibody class and antibody subclass) and in vitro anti-bacterial activity of meningococcal antibodies. The antibody sets represented the human isotypes IgG1, IgG3 and IgM, which dominate during immune response against protein antigens. The binding activities were quite similar for all these isotypes, surprisingly also for the pentameric IgM. Interestingly, monomeric IgM, prepared from pentameric IgM by partially reduction and alkylation, had similar binding activities as the original pentameric IgM. Regarding in vitro anti-bacterial activity, chIgG1 was superior in SBA (serum bactericidal activity) compared with chIgG3, while chIgG3 was more efficient in OP (opsonophagocytosis; measured by flow cytometry) than chIgG1. ChIgM showed slightly higher SBA than chIgG1 on molar basis, and much higher OP than chIgG3 and chIgG1. A lower concentration of antibodies was needed against the P1.16 than against the P1.7 epitope to induce SBA, but this was not the case for OP.


title: The triple power of D³: protein intrinsic disorder in degenerative diseases.

text: This review is an update of an article published four years ago (Uversky V.N. (2009) Intrinsically disordered proteins in neurodegenerative diseases: another illustration of the D² concept. Frontiers in Bioscience 14, 5188-5238). The major goal of this review is to show the interconnections between intrinsically disordered proteins (IDPs) and human neurodegeneration. This brings to existence a new D³ concept: protein intrinsic Disorder in neuroDegenerative Diseases. An important aspect of the D³ concept is that it deals with three D³'s, emphasizing that intrinsically Disordered proteins are abundantly found in various neuroDegenerative Diseases (the first D³), that these IDPs provoke neuroDegeneration due to their Dysfunctionality (the second D³), and that neuroDegeneration-related IDPs are often controlled by other Disordered proteins (the third D³).


title: Etiologic study of upper respiratory infections of household dogs.

text: Infectious tracheobronchitis (ITB), also known as the kennel cough, is a respiratory syndrome of dogs and usually appears to be contagious among dogs housed in groups. Etiologic agent of ITB is multiple and sometimes complex. In the present study, 68 household dogs showing clinical signs of respiratory infection were examined, and 20 dogs (29.4%) were found to be positive for either of following agents. Bordetella bronchiseptica (B.b.) was most frequently detected from nasal and oropharynx sites of 7 dogs (10.3%). Among the viruses examined, canine parainfluenza virus (CPIV) was detected with the highest frequency (7.4%). Other pathogens included in the order of frequency group 1 canine coronavirus (4.4%), canine adenovirus type 2 (2.9%), group 2 canine respiratory coronavirus (1.5%), and canine distemper virus (1.5%). Only 2 cases showed mixed infections. Neither influenza A virus nor canine bocavirus (minute virus of canines) was found in any dogs examined. These results indicate that both B.b. and CPIV are likely to be the principal etiologic agents of canine ITB in Japan, and they may be considered as the target for prophylaxis by vaccination.


title: [Study on the epidemiology and measures for control on severe acute respiratory syndrome in Guangzhou city].

text: OBJECTIVE To analyze the epidemiological characteristics, related risk factors, measures for its control of severe acute respiratory syndrome (SARS). METHODS Data on epidemiological features, pathogens and measures for control were collected and analyzed. RESULTS Since Jan 2003, infectious atypical pneumonia (AP) has become epidemic in Guangzhou city. The first autochthonous case was identified on Jan 2nd. Number of cases started to increase since February and reached peak in the early 10 days of February. Hereafter the epidemic tended to decline in March and since early April, the average number of new cases began to decrease, less than 10 per day. Epidemiological studies revealed that the number of cases aged between 20 and 50 was higher than that below the age of 20. Of the total 966 cases, 429 were males versus 537 females. Geographically, the epidemics covered all 13 districts of Guangzhou, but 95% of the cases concentrated in 7 urban districts. As for professional distribution, health care workers accounted for 28.67% of the total cases. There were 36 deaths, aged from 5 to 89, with half of them older than 60. Out of the victims, 38.9% of them had complications as hypertension, diabetes, heart diseases and COPD etc. Data regarding the clustering features of cases showed that there were 42 families having 2 or more cases in one family, while 277 health workers suffered from SARS were concentrated in 28 hospitals. Only one outbreak took place in a public setting but no outbreak was reported in schools. Relevant research also indicated that SARS could be classified as an air-borne infectious disease, transmitted through aerosol and droplets, but close contact also played an important role in the mode of transmission. The disease was highly infectious, suggesting that people who had close contact with patients in the place with poor ventilation was in greater risk of getting infection. The incubation period ranged from 1 to 11 days (mainly from 3 to 8 days), with an average of 5 days. According to our observation, the following measures might be effective such as: early diagnosis, isolation and treatment provided to the patients, and suspected cases under medical observation should also be put in separate places. Improving ventilation and regular disinfection over air and stuff in hospital wards were also recommended. In order to prevent iatrogenic infection, sense on self-protection among health care workers must be strengthened. Patients were not allowed to be visited by any one other than hospital staff. CONCLUSION SARS is a preventable disease and can be under control. It is of great importance to prevent clustered SARS cases and the prevention of iatrogenic infection is essential.


title: Characterization of monoclonal antibodies to the hemagglutinin-esterase glycoprotein of a bovine coronavirus associated with winter dysentery and cross-reactivity to field isolates.

text: Seven hybridoma cell lines producing monoclonal antibodies (MAbs) to the hemagglutinin-esterase (HE) glycoprotein of bovine coronavirus (BCV) were obtained from BALB/c mice that were immunized with an enriched peplomeric fraction of the winter dysentery (WD)-associated strain BCQ.2590. The specificities of these MAbs to either the dimeric (140-kDa) or the monomeric (65-kDa) form of the HE glycoprotein were determined by Western immunoblotting experiments with purified virus and immunoprecipitation tests with [35S]methionine-labelled infected cell extracts. Four of these anti-HE MAbs inhibited the hemagglutinating activity of the virus and three weakly neutralized its infectivity in vitro. In addition, competition binding assays allowed for the definition of two independent antigenic domains (domains A and D) and two overlapping antigenic domains (domains B and C) for the HE glycoprotein of the WD-associated strain; epitopes located within antigenic domain A were not associated with hemagglutination inhibition (HAI) and virus neutralization activities. In HAI tests, the four anti-HA MAbs defined two distinct antigenic subgroups among 24 BCV field isolates that have been associated with either typical outbreaks of WD or neonatal calf diarrhea (NCD) in Quebec dairy herds from 1986 to 1996. The Quebec WD-associated strains of BCV, as well as some of the NCD-associated strains isolated since 1991, fell within a subgroup distinct from that of the prototype Mebus strain.


title: A systematic review of the association between circulating concentrations of C reactive protein and cancer.

text: The objective of this study was to review and summarise the published evidence for an association between circulating concentrations of C reactive protein (CRP) and cancer through a systematic review. 90 discrete studies were identified. 81 (90%) were prevalent case-control or cross-sectional studies, and only 9 studies had a prospective design. In most prevalent studies, CRP concentrations were found to be higher in patients with cancer than in healthy controls or controls with benign conditions. Of the nine large prospective studies identified in this review, four reported no relationship between circulating CRP levels and breast, prostate or colorectal cancers, and five studies found that CRP was associated with colorectal or lung cancers. Most of the studies evaluating CRP as a diagnostic marker of cancer did not present relevant statistical analyses. Furthermore, any association reported in the prevalent studies might reflect reverse causation, survival bias or confounding. The prospective studies provided no strong evidence for a causal role of CRP in cancer. Instead of further prevalent studies, more large prospective studies and CRP gene-cancer association studies would be valuable in investigating the role of CRP in cancer.


title: Information management during mass casualty events.

text: Preparing for a mass casualty event starts long before the event. Being able to provide care during an event requires collecting information about processes, equipment, supplies, and personnel to anticipate different situations. Planning should include an impact analysis to identify and prioritize critical services that need to be maintained. Impact analysis is carried out in concert with the entire health care delivery organization, not by individual departments. Planning may include addressing inadequate staffing, disrupted supply chains, and loss of information systems. Care may need to be provided in alternate locations, without access to the usual information resources. Mass casualty events also depend on the availability of communications to inform others on the emergency response teams, as well as to inform patients and the public.


title: Protection of chickens after live and inactivated virus vaccination against challenge with nephropathogenic infectious bronchitis virus PA/Wolgemuth/98.

text: Protection provided by live and inactivated virus vaccination against challenge with the virulent nephropathogenic infectious bronchitis virus (NIBV) strain PA/Wolgemuth/98 was assessed. Vaccinations with combinations of live attenuated strains Massachusetts (Mass) + Connecticut (Conn) or Mass + Arkansas (Ark) were given by eyedrop to 2-wk-old specific-pathogen-free leghorn chickens. After live infectious bronchitis virus (IBV) vaccination, some chickens at 6 wk of age received an injection of either an oil emulsion vaccine containing inactivated IBV strains Mass + Ark or an autogenous vaccine prepared from NIBV PA/Wolgemuth/98. Challenge with PA/Wolgemuth/98 was given via eyedrop at 10 wk of age. Serum IBV enzyme-linked immunosorbent assay antibody geometric mean titers (GMTs) after vaccination with the combinations of live attenuated strains were low, ranging from 184 to 1,354, prior to NIBV challenge at 10 wk of age. Both inactivated vaccines induced an anamnestic response of similar magnitudes with serum GMTs of 6,232-12,241. Assessment of protection following NIBV challenge was based on several criteria virus reisolation from trachea and kidney and renal microscopic pathology and IBV-specific antigen immunohistochemistry (IHC). Live attenuated virus vaccination alone with combinations of strains Mass + Conn or Mass + Ark did not protect the respiratory tract and kidney of chickens after PA/Wolgemuth/98 challenge. Chickens given a live combination vaccination of Mass + Conn and boosted with an inactivated Mass + Ark vaccine were also susceptible to NIBV challenge on the basis of virus isolation from trachea and kidney butshowed protection on the basis of renal microscopic pathology and IHC. Live IBV-primed chickens vaccinated with an autogenous inactivated PA/Wolgemuth/98 vaccine had the highest protection against homologous virulent NIBV challenge on the basis of virus isolation.


The consequences of fear.


title: Etiology and management of pharyngitis and pharyngotonsillitis in children: a current review.

text: Although viruses are the most common causes of childhood throat infections, interest in the etiology of these infections has primarily focused on whether an individual episode is caused by group A beta-hemolytic streptococcus (GABHS), particularly since the recent outbreaks of rheumatic fever in certain areas of the country. Penicillin remains the cornerstone of treatment in GABHS pharyngitis. Early treatment effects prompt clinical improvement and reduces the risk of transmission. Whether early treatment suppresses immunologic response and results in a higher recurrence rate than does delayed treatment is still unknown, but recent evidence suggests that it does not. The causes of persistent GABHS carriage, its clinical importance, and optimal methods of treatment are all still in question. When penicillin treatment does not eradicate carriage, other drugs may be efficacious. In children severely affected with recurrent throat infection, tonsillectomy is generally effective and is sometimes a desirable option.


title: Molecular analysis of the S glycoprotein gene of bovine coronaviruses isolated in Japan from 1999 to 2006.

text: In total, 55 isolates of Bovine coronavirus (BCoV) were collected from cases of enteric and respiratory disease occurring between 1999 and 2006 in Japan. Phylogenetic analysis of the polymorphic region of the S glycoprotein gene of these isolates, together with those of other known strains, classified the BCoV strains and isolates into four clusters. Recent field isolates display distinctive genetic divergence from the prototype enteric BCoV strains--Mebus, Quebec, Kakegawa, F15 and LY138--and have diverged in three different aspects over 8 years. These data suggested that the genetic divergence in the polymorphic region of the S glycoprotein has progressed considerably; thus, molecular analysis of this region should be useful in investigating the molecular epidemiology of BCoV. In addition, based on the differences in amino acids among the isolates, our study did not reveal the presence of certain genetic markers of pathogenicity and clinical symptoms in this polymorphic region.


Acute laryngitis superimposed on chronic laryngitis.


title: Microbicides and the environmental control of nosocomial viral infections

text: Abstract Viruses are important causes of acute and chronic diseases in humans. Newer viruses are still being discovered and those that are already known are being incriminated in the aetiology of clinical conditions with hitherto unknown causes. Apart from frequently causing infections in the general community, many types of viruses are also significant nosocomial pathogens. While it is generally agreed that we underestimate the proportion of nosocomial infections that are viral, due to a lack of routine monitoring, viruses easily account for more than 30% of the cases of hospital-acquired infections in many paediatric settings. Indeed, the relative importance of viruses in this respect is increasing due to a number of societal and demographic changes as well as alterations in healthcare practices. Safe vaccines against many common nosocomial viral agents are currently unavailable while there is also a virtual lack of effective and affordable chemotherapy against them. There is, therefore, renewed emphasis on preventive strategies by better understanding of the relative importance of various vehicles in the nosocomial spread of viruses and by infection control using microbicides. This, in turn, has stimulated considerable interest in the development of formulations that are not only safer but which also have demonstrated activity against major types of nosocomial viral pathogens. Further, much work is now underway to design better methods to assess the virucidal activity of microbicides used to decontaminate hands, reusable medical devices and environmental surfaces in critical areas of healthcare settings. It is anticipated that these approaches will result in reducing the health and economic impact of nosocomial infections due to viruses.


title: The region between the M and S genes of porcine haemagglutinating encephalomyelitis virus is highly similar to human coronavirus OC43.

text: The nucleotide sequences of the regions between the membrane and spike protein genes of three strains of porcine haemagglutinating encephalomyelitis virus (HEV) were determined. A total of 739 (HEV strain 67N) and 751 (strains NT9 and VW572) nucleotides were sequenced. Two ORFs, potentially encoding proteins of 12.8 and 9.6 kDa, were identified. Pairwise comparisons with the corresponding ORFs in bovine coronavirus (BCV) and human coronavirus (HCV) OC43 revealed sequence similarities of greater than 88.5 percent at the nucleotide and 85.3 percent at the amino acid level for the 12.8 kDa ORF product. For the 9.6 kDa ORF product similarities were greater than 96.9 percent and 95.2 percent, respectively. An additional 12 nucleotide deletion upstream of the 12.8 kDa ORF start codon was found in HEV 67N compared to NT9 and VW572. These results reveal a genomic organization of HEV in the region analysed that is homologous to HCV OC43 but different from BCV.


title: Remission of diarrhoea due to cryptosporidiosis in an immunodeficient child treated with hyperimmune bovine colostrum.

text: A boy aged 6 months who presented with poor weight gain, diarrhoea, and infection with Pneumocystis carinii was found to have congenital hypogammaglobulinaemia, which did not improve despite monthly treatment with intravenous gammaglobulin. At the age of 3 years and 2 months he developed severe vomiting and diarrhoea due to cryptosporidiosis, which failed to respond to conventional treatment. Infusion of hyperimmune bovine colostrum produced against parasite antigen, given by nasogastric tube, was started after symptoms had persisted for three weeks. His vomiting and diarrhoea resolved within five days of treatment, and oocysts were no longer seen in the stools after eight days. Later, however, he developed a rare complication, and oocysts were found in the common bile duct. Hyperimmune bovine colostrum may be useful in the treatment of many patients with immunodeficiency disorders.


Biologists lobby China's government for funding reform.


Coronavirus antibody titres in cats in New Zealand.


title: Borna disease virus.

text: Borna disease virus, a negative-strand RNA virus, infects a wide variety of warm-blooded animals. Depending on the age of the host and the integrity of its immune response, infection may be asymptomatic or cause a broad spectrum of behavioral disorders. Unusual features of Borna disease virus biology include nuclear localization of replication and transcription; diverse strategies for regulation of gene expression; and interaction with signaling pathways resulting in subtle neuropathology. Although the question of human infection remains unresolved, burgeoning interest in this unique pathogen has provided tools for exploring the pharmacology and neurochemistry of neuropsychiatric disorders potentially linked to infection. Analysis of rodent models of infection has yielded insights into mechanisms by which neurotropic agents and/or immune factors may impact developing or mature central nervous system circuitry to effect complex disturbances in movement and behavior.


title: Structural studies of algal lectins with anti-HIV activity.

text: A number of antiviral lectins, small proteins that bind carbohydrates found on viral envelopes, are currently in pre-clinical trials as potential drugs for prevention of transmission of human immunodeficiency virus (HIV) and other enveloped viruses, such as the Ebola virus and the coronavirus responsible for severe acute respiratory syndrome (SARS). Lectins of algal origin whose antiviral properties make them candidate agents for prevention of viral transmission through topical applications include cyanovirin-N, Microcystis viridis lectin, scytovirin, and griffithsin. Although all these proteins exhibit significant antiviral activity, their structures are unrelated and their mode of binding of carbohydrates differs significantly. This review summarizes the current state of knowledge of the structures of algal lectins, their mode of binding of carbohydrates, and their potential medical applications.


title: Severe febrile respiratory illnesses as a cause of mass critical care.

text: Febrile respiratory illnesses with respiratory failure are one of the most common reasons for admission to the intensive care unit. Most causes of febrile respiratory illness are bacterial and viral agents of community-acquired pneumonia. However, a small number of rare and highly contagious agents can initially present as febrile respiratory illnesses, which can lead to an epidemic that can greatly impact the health care system. This impact includes sustained mass critical care, with potential scarcity of critical resources (eg, positive-pressure ventilators), spread of disease to health care workers, sustained spread within the community, and extensive morbidity and mortality. The main agents of febrile respiratory illness that would lead to an epidemic include influenza, the coronavirus that causes severe acute respiratory syndrome, smallpox, viral hemorrhagic fever, plague, tularemia, and anthrax. Recognition of these agents occurs largely based on epidemiological clues, and management consists of antibiotics, antivirals, supportive care, and positive-pressure ventilation. Acute respiratory failure and acute respiratory distress syndrome occur with these agents, so a lung-protective (low tidal volume) ventilation strategy is indicated. Additional respiratory care measures, such as nebulized medications, bronchoscopy, humidified oxygen, and airway suctioning, potentiate aerosolization of the virus or bacteria and increase the risk of transmission to health care workers and patients. Thus, appropriate personal protective equipment, including an N95 mask or powered air-purifying respirator, is indicated. A basic understanding of the epidemiology, clinical findings, diagnosis, and treatment of these agents will provide a foundation for early isolation, evaluation, infection control, and public health involvement and response in cases of a febrile respiratory illness that causes respiratory failure.


Expression library immunization: a road map for discovery of vaccines against infectious diseases.


title: Cloning of a human parvovirus by molecular screening of respiratory tract samples.

text: The identification of new virus species is a key issue for the study of infectious disease but is technically very difficult. We developed a system for large-scale molecular virus screening of clinical samples based on host DNA depletion, random PCR amplification, large-scale sequencing, and bioinformatics. The technology was applied to pooled human respiratory tract samples. The first experiments detected seven human virus species without the use of any specific reagent. Among the detected viruses were one coronavirus and one parvovirus, both of which were at that time uncharacterized. The parvovirus, provisionally named human bocavirus, was in a retrospective clinical study detected in 17 additional patients and associated with lower respiratory tract infections in children. The molecular virus screening procedure provides a general culture-independent solution to the problem of detecting unknown virus species in single or pooled samples. We suggest that a systematic exploration of the viruses that infect humans, "the human virome," can be initiated.


title: [Comparison of clinical features of severe acute respiratory syndrome among different transmission generations].

text: OBJECTIVE To investigate the differences among various transmission generation of severe acute respiratory syndrome (SARS) by comparing the corresponding clinical data. METHODS The clinical data of 84 patients with SARS were retrospectively studied, 66 women and 18 men, mean age (29.2 +/- 10.3) years old, 96.4% of whom were health care workers. All subjects had exposure to a source patient and the epidemic progressively propagated in a short time. For the infectious chain, we defined these patients who had exposure to a source patient as the primary cases, which included 35 patients (41.7%). Patients who got the disease by exposure to the primary cases were defined as secondary cases, which included 34 patients (40.5%). Similarly, the tertiary cases included 15 patients (17.9%). RESULTS (1) No statistical differences in age, sex ratio, incubation period and hospitalization duration among various infectious generations were found (P > 0.05). (2) With descending in infectious generations, the initial temperature lowered, and cases with cough decreased (P < 0.05) with no statistical differences in the peak temperature, other accompanying symptoms and leukopenia (P > 0.05). (3) With descending in infectious generations, the course from the appearance of pulmonary lesions to their resolution shortened (P < 0.05). No differences were found in the maximal involved pulmonary fields, duration from initial fever to appearance of pulmonary lesions and course from the initial pulmonary lesions to their peak among the above three generations (P > 0.05). (4) No statistical differences were found in ways of oxygen therapy and classes of antibiotics prescribed (P > 0.05). With descending in infectious generation, cases treated with methylprednisolone, human gamma-immunoglobulin, interferon-alpha, and antiviral drugs (oral ribavirin or oseltamivir) increased (P < 0.05) and the duration of their use also increased (P < 0.05). CONCLUSIONS With descending in infectious generations, the clinical features of SARS may become ameliorated.


title: Characterization and treatment of cidofovir-resistant vaccinia (WR strain) virus infections in cell culture and in mice.

text: The wild-type (WT) vaccinia (WR strain) virus is highly virulent to mice by intranasal inoculation, yet death can be prevented by cidofovir treatment. A cidofovir-resistant (CDV-R) mutant of the virus was developed by 15 Vero cell culture passages in order to determine cross-resistance to other inhibitors, growth characteristics, virulence in infected mice, and suitability of the animal model for studying antiviral therapies. Comparisons were made to the original WT virus and to a WT virus passaged 15 times in culture (WTp15 virus). Cidofovir inhibited WT, WTp15, and CDV-R viruses by 50% at 61, 56 and 790 microM, respectively, in plaque reduction assays, with similar inhibition seen in virus yield studies. Cross-resistance occurred with compounds related to cidofovir, but not with unrelated nucleosides. The resistant virus produced 300-fold fewer infectious particles (PFU) than WT and WTp15 viruses in mouse C1271 cells, yet replicated similarly in Vero (monkey) cells. The CDV-R virus was completely attenuated for virulence at 10(7) PFU per mouse in normal BALB/c mice and in severe combined immunodeficient (SCID) mice. The WTp15 virus was 100-fold less virulent than WT virus in BALB/c mice. Thus, the lack of virulence of the resistant virus in the animal model is explained partly by its reduced ability to replicate in mouse cells and by attenuation occurring as a result of extensive cell culturing (inferred from what occurred with the WTp15 virus). Lung and snout virus titre reduction parameters were used to assess antiviral activity of compounds in BALB/c mice infected intranasally with the CDV-R virus. Cidofovir, HDP-cidofovir and arabinofuranosyladenine treatments reduced lung virus titres or = eight-fold. The animal model appears to have limited utility in drug efficacy testing.


title: In-vivo assessment of barbed suturing thread with regard to tissue reaction and material absorption in a rat model.

text: AIM The laparoscopic approach in urological surgery demands a high degree of skill in intracorporeal suturing and knot tying. In an effort to reduce the amount of time required to perform a suture, new materials have been developed that through selfanchorage distribute tension more evenly across the suture and also eliminate the need of knot tying. The goal of this study was to assess the in vivo tissue response to a novel material (V-Loc tm; Covidien) in comparison to established materials (Vicryl, PDS II), in the case of bladder suturing, in a rat model. METHODS The study included 48 male Wistar rats. All underwent a median abdominal incision, with a 1cm cystotomy, followed by a running suture. The suture material used was either V-Loc absorbable self anchoring thread, Vicryl threaded absorbable suture or monofilament absorbable suture. The abdominal cavity and the bladder suture were macroscopically evaluated at the rats' scheduled death at 3 and 6 weeks. The bladder wall was microscopically assessed by a pathologist, with regard to tissue reaction and suture material degradation. RESULTS All rats survived the procedure, with the abdominal scar fully healed at week 2. There were no signs of infection or lithiasis during the observation. Macroscopically, at 3 weeks, the suture material was recognizable and visible in all cases, with special mention that the V-Loc thread was considerably more rigid, retaining its shape almost entirely, and provoked more adhesion of the surrounding tissue. At 6 weeks, the suture was indistinguishable in the bladder wall in the case of monofilament absorbable material, barely visible in the case of Vicryl, while the aspect of the V-Loc suture resembled the one at 3 weeks, with the material still clearly visible in the bladder wall, shape almost entirely maintained, and surrounding tissue adherence. Microscopically, at 3 weeks and 6 weeks, all bladder walls examined had regained their structure. At 3 weeks, the monofilament absorbable suture showed intense tissue reaction, with the material already in phagocytosis; at 6 weeks, no clear evidence of leftover material was observed. At 3 weeks, the Vicryl material showed moderate tissue reaction, with phagocytosis initiated between the strands of the material; at 6 weeks, the material was almost entirely absorbed, but with a clear leftover tissue reaction. In the case of the V-Loc suture, due to the hardness of the thread, the material itself could not be cut for analysis with the bladder wall, and the examination could only involve the bladder wall and marks of the thread. Thus, the tissue reaction was minimal, as was the presence of phagocytes at the suture site. The material showed little, if any, signs of absorption after 6 weeks. CONCLUSION The materials tested all proved equally effective in suturing the bladder wall in a rat model. However, the novel barbed thread proved a consistently low in-vivo absorption rate, while maintaining its rigidity over time. More research is needed to assess the possible clinical implications of these findings.