title: Severe acute respiratory syndrome: an update.

text: PURPOSE OF REVIEW An international outbreak of severe acute respiratory syndrome, a recently recognized syndrome caused by the newly identified severe acute respiratory syndrome-associated coronavirus, began in November 2002 and ended in July 2003. Since then, a large body of research on the syndrome has been published; the most updated developments are summarized here. RECENT FINDINGS Recent findings suggest that animal severe acute respiratory syndrome-like coronaviruses may have been transmitted to humans without detection for years before the recent outbreak, and that such transmission may be continuing today. The 2002-2003 outbreak probably originated from similar animal-to-human transmission, but selection and purification of the animal severe acute respiratory syndrome-like virus appears to have occurred, creating the more virulent severe acute respiratory syndrome-associated coronavirus. Recent studies have documented that severe acute respiratory syndrome-associated coronavirus is primarily transmitted via contact and/or respiratory droplets and that the combination of standard, contact, and droplet precautions is generally effective for its control. It has been shown that severe acute respiratory syndrome-associated coronavirus is typically relatively inefficiently transmitted, with the notable exception of transmission during superspreading events. Insights into the pathogenesis of severe acute respiratory syndrome have been made: one study suggests that human leukocyte antigen HLA-B*4601 is a possible risk factor for more severe disease, while another identifies angiotensin-converting enzyme 2 as a cellular receptor for severe acute respiratory syndrome-associated coronavirus. Promising treatments have been identified, including interferons, an anti-spike monoclonal antibody, and fusion inhibitors. In addition, many promising vaccines are currently in development. SUMMARY New findings regarding severe acute respiratory syndrome are continuing to be discovered at an unprecedented pace, permitting a better understanding of the disease and enabling better preparation for its possible return.


title: Dynamic Modeling COVID-19 for Comparing Containment Strategies in a Pandemic Scenario.

text: The emergence of coronavirus disease 2019 (COVID-19) in the United States has forced federal and local governments to implement containment measures. Moreover, the severity of the situation has sparked engagement by both the research and clinical community with the goal of developing effective treatments for the disease. This article proposes a time dynamic prediction model with competing risks for the infected individual and develops a simple tool for policy makers to compare different strategies in terms of when to implement the strictest containment measures and how different treatments can increase or suppress infected cases. Two types of containment strategies are compared: (1) a constant containment strategy that could satisfy the needs of citizens for a long period; and (2) an adaptive containment strategy whose strict level changes across time. We consider how an effective treatment of the disease can affect the dynamics in a pandemic scenario. For illustration we consider a region with population 2.8 million and 200 initial infectious cases assuming a 4% mortality rate compared with a 2% mortality rate if a new drug is available. Our results show compared with a constant containment strategy, adaptive containment strategies shorten the outbreak length and reduce maximum daily number of cases. This, along with an effective treatment plan for the disease can minimize death rate.


title: Computer-assisted functional assessment and documentation.

text: Occupational therapy personnel spend considerable time on functional assessment and documentation. The rapid and error-free information processing ability of computers offers methods for data collection, clinical decision making, and reporting of functional assessment results, which may be more effective and efficient than the current paper-and-pencil methods. Computers can provide assistance throughout the many steps of the functional assessment and reporting process. Software is available to assist in direct data collection, data reduction, and analysis, or to assess complex data systems and databases. Additionally, computers can use assessment techniques, such as dynamic question sets and expert systems, which have not been possible with paper-and-pencil methods. Furthermore, data collected on computers are readily available to be integrated in computer-assisted report writing and can be compiled into large databases for analysis. These databases can be used for program evaluation, continuous quality improvement exercises, and other research objectives. This paper provides a snapshot of these computer uses in the assessment process.


title: CT features of coronavirus disease 2019 (COVID-19) pneumonia: experience of a single center in Southern Italy.

text: PURPOSE The aim of this study was to report the radiological features of chest CT scan of patients with coronavirus disease 2019 (COVID-19) living in a town in Southern Italy where a significant outbreak of the disease occurred. METHODS We revised the CT scan of 62 patients (34 male, 28 female, mean age 71 +/- 14 years) with clinical and laboratory signs of COVID-19, as assessed by positive SARS-CoV-2 RT-PCR testing. All patients underwent chest CT at the time of admission to the hospital. A semi-quantitative scoring system was used to evaluate the extension of the disease. RESULTS Out of the 62 patients the main radiological findings were reticular pattern (29%), ground-glass opacities (24%), crazy paving pattern (11%) and consolidation (35%). Most of the lesions were bilateral (97%), posterior (95%) and located near pleura (50%) or lung fissures (45%), mainly involving the lower right lobe (56%) and lower left lobe (23%). Pleural thickening was observed in 72.6% of patients and pleural effusion in 18%. Median value of the score was 7.0 and was significantly higher in male than female (8.5 vs 6.0, p=0.03) and in patients with pleural thickening compared to those without this finding (8.0 vs 5.0, p=0.03).


title: [A modification of the ABC method (avidin-biotin-peroxidase complex) for the detection of viral antigens in coronavirus infections in cats (FIP) and parvovirus type 2 infections in dogs].

text: A modified ABC-method (avidin-biotin-peroxidase complex) is established to demonstrate viral antigens in paraffin sections of tissues of coronavirus (FIP) infected cats and parvovirus type 2 infected dogs using monoclonal antibodies. The pros and cons and potential sources of error are described and discussed.


title: Synthesis, antiviral and cytotoxic investigation of 2-phenyl-3-substituted quinazolin-4(3H)-ones.

text: OBJECTIVES A series of 3(benzylideneamino)-2-phenyl quinazoline-4(3H)-ones was synthesized by reaction of 3-amino-2-phenyl-3H-quinazoline-4-one with various carbonyl compounds. MATERIALS AND METHODS Chemical structures of the synthesized compounds were confirmed by IR, 1H-NMR and mass spectral analysis. Title compounds were investigated for cytotoxicity and antiviral activity against herpes simplex virus-1 (KOS), herpes simplex virus-2 (G), vaccinia virus, vesicular stomatitis virus, herpes simplex virus-1 TK-KOS ACVr, para influenza-3 virus, reovirus-1, Sindbis virus, Coxsackie virus B4, Punta Toro virus, feline corona virus (FIPV), feline herpes virus, respiratory syncytial virus, influenza A H1 N1 subtype, influenza A H3N2 subtype, influenza B and vesicular stomatitis virus. RESULTS AND CONCLUSION Compound 3d was found inhibit viral replication of para influenza-3virus, reovirus-1, Sindbis virus, Coxsackie virus B4, Punta Toro virus in Vero cell cultures.


PEDV leader sequence and junction sites.


title: The laboratory diagnosis of severe acute respiratory syndrome: emerging laboratory tests for an emerging pathogen.

text: The 2003 pandemic of Severe Acute Respiratory Syndrome (SARS) profiled the ability of modern diagnostic microbiology and molecular biology to identify, isolate and characterize, within weeks, a previously unknown viral infectious pathogen. The culprit, SARS coronavirus (SARS-CoV), was detected in patient specimens by traditional cell culture using an unusual cell line for respiratory viruses, Vero E6, and by reverse transcriptase polymerase chain reaction (RT-PCR) targeting the polymerase 1 B region of the genome. In addition, serologic assays were rapidly developed, and the genome of this large virus was sequenced within one month of its spread to North America. At the present time, diagnostics have progressed to the point that RT-PCR has a sensitivity approaching 80% within the first few days of onset of illness, while serology has a sensitivity close to 100% on convalescent sera taken >21 days after illness onset. Viral culture remains a method confined to biosafety level III laboratories. The specificity of RT-PCR and serology remains to be conclusively defined, but in most studies to date seems to be >90%. Serologic cross-reactivity with human coronaviruses causing the common cold may be a problem with some serologic assays. The early development of SARS-CoV diagnostics is now being replaced by refinement and optimization of these assays. Although at the present time we do not have a test that will definitively rule in or rule out SARS at the time of initial presentation of a patient with a respiratory infection, modifications of existing assays will hopefully result in our ability to make this diagnosis with a high degree of accuracy in the future.


title: Evaluation of right versus left laparoscopic donor nephrectomy.

text: OBJECTIVES With the advent of laparoscopic donor nephrectomy, there has been a general underuse of right laparoscopic donor nephrectomy versus left because of concerns regarding higher complication rates and poorer outcomes. We performed a retrospective analysis of our laparoscopic donor nephrectomy series with an emphasis on the side of the kidney retrieved and the outcomes of donors and recipients. MATERIALS AND METHODS Data on 94 consecutive donor-recipient pairs (188 patients) were reviewed. All donor nephrectomies were performed by pure laparoscopy. There were 74 left laparoscopic donor nephrectomies and 20 right laparoscopic donor nephrectomies. Intraoperative parameters and graft outcome were recorded and the data were analyzed to compare right laparoscopic donor nephrectomy versus left laparoscopic donor nephrectomy using a computer software system. Follow-up ranged from 1 to 6 years (mean, 3.4 y). RESULTS There were no significant differences in any intraoperative or postoperative parameters, except in a slightly higher warm ischemia time in right laparoscopic donor nephrectomy versus left laparoscopic donor nephrectomy, but this did not translate into an adverse effect on renal recovery. Acute graft rejection was observed in 2 of 74 patients who had a left laparoscopic donor nephrectomy (2.7%) and none of the patients who had a right laparoscopic donor nephrectomy. Chronic graft loss was observed in 2 of 74 patients who had a left laparoscopic donor nephrectomy (2.7%); 1 of recurrent pyelonephritis and sepsis and 1 renal oxalosis. No graft losses were observed in any patient who had a right laparoscopic donor nephrectomy. Mean serum creatinine levels in recipients at 1, 3, 6, 9, and 12 months were equivalent for right laparoscopic donor nephrectomy versus left laparoscopic donor nephrectomy; they were 120, 110, 110, 110, 110 µmol/L, versus 110, 110, 110, 110, 110 µmol/L. (1.35, 1.21, 1.24, 1.21, 1.26 mg/dL versus 1.22, 1.17, 1.17, 1.17, 1.23 mg/dL). CONCLUSIONS This single center study demonstrates equivalent results with left laparoscopic donor nephrectomy and right laparoscopic donor nephrectomy with no adverse effects of right laparoscopic donor nephrectomy on donor-recipient outcome or renal function.


title: [Prokaryotic expression and characterization of two recombinant receptor-binding domain(RBD) proteins of human coronavirus NL63(HcoV-NL63)].

text: The receptor-binding domain(RBD) protein of HCoV-NL63 is a major target in the development of diagnostic assay and vaccine, it has a pivotal role in receptor attachment, viral entry and membrane fusion. In this study, we prepared 2 purified recombinant HCoV-NL63 RBD proteins using in E. coli system and identified the proteins by Western blotting. We first optimized codon and synthesized the RL (232-684aa)coding gene, then amplified the RL or RS(476-616aa) coding gene via PCR using different primers . The RL or RS coding gene was cloned into the pM48 expression vector fused with TrxA tag. The RBD (RL and RS) of HCoV-NL63 were expressed majorly as inclusion body when expressed in E. coli BL21pLys S under different conditions. The expressed products were purified by affinity chromatography then analyzed by SDS-PAGE and Western blotting. Our results showed that the recombinant RBD proteins were maximally expressed at 37 degrees C with 0. 8mM IPTG induction for 4h. RL or RS protein with 95 % purity was obtained and reacted positively with anti-sera from mice immunized with the recombinant vaccinia virus (Tiantan strain) in which HCoV-NL63 RL or RS protein was expressed. In conclusion, the purified recombinant RBD proteins(RL and RS)derived from E. coli were first prepared in China and they might provide a basis for further exploring biological role and vaccine development of HCoV-NL63.


title: Drug targets for COVID-19 therapeutics: Ongoing global efforts.

text: The current global pandemic COVID-19 caused by the SARS-CoV-2 virus has already inflicted insurmountable damage both to the human lives and global economy. There is an immediate need for identification of effective drugs to contain the disastrous virus outbreak. Global efforts are already underway at a war footing to identify the best drug combination to address the disease. In this review, an attempt has been made to understand the SARS-CoV-2 life cycle, and based on this information potential druggable targets against SARS-CoV-2 are summarized. Also, the strategies for ongoing and future drug discovery against the SARSCoV- 2 virus are outlined. Given the urgency to find a definitive cure, ongoing drug repurposing efforts being carried out by various organizations are also described. The unprecedented crisis requires extraordinary efforts from the scientific community to effectively address the issue and prevent further loss of human lives and health.


title: Absence of porcine interferon alpha secreting cells in severe combined immunodeficiency (SCID) mice inoculated with porcine leukocytes.

text: A low frequency leukocyte subpopulation, referred to as natural interferon producing cells (NIPC) is able to produce high amounts of interferon alpha (IFN-alpha) following contact with noninfectious viral structures. In order to examine the possible leukocytic nature and bone marrow origin of NIPC, severe combined immunodeficiency (SCID) mice were reconstituted with porcine leukocyte populations, including bone marrow cells. At different times after reconstitution, enriched CD4 and CD45 positive porcine cells were isolated from various mouse organs and tested for the presence of porcine NIPC by porcine IFN-alpha specific ELISPOT assay, after in vitro stimulation by UV inactivated transmissible gastroenteritis virus (TGEV). Although engraftment of porcine cells in SCID mice was shown by flow cytometry and by the production of pig immunoglobulins, no IFN-alpha secreting cells could be detected. This result suggests that NIPC do not derive from bone marrow precursor cells, or that growth factors needed for in vivo expansion of porcine NIPC were absent in mice.


title: Factors affecting perceived risk of contracting severe acute respiratory syndrome among academic physicians.

text: SARS disproportionately affects healthcare providers. A physician survey was administered within three hospitals providing care to SARS patients. Knowing a colleague who contracted SARS and previous occupational exposure to infectious agents were significantly predictive of greater perceived risk, whereas perceived effectiveness of precautions and provision of direct care were not.


Challenges and opportunities of public health research in Nepal.


APIC guide highlights role of ambulatory care in disaster response.


title: Guidelines for preventing health-care--associated pneumonia, 2003: recommendations of CDC and the Healthcare Infection Control Practices Advisory Committee.

text: This report updates, expands, and replaces the previously published CDC "Guideline for Prevention of Nosocomial Pneumonia". The new guidelines are designed to reduce the incidence of pneumonia and other severe, acute lower respiratory tract infections in acute-care hospitals and in other health-care settings (e.g., ambulatory and long-term care institutions) and other facilities where health care is provided. Among the changes in the recommendations to prevent bacterial pneumonia, especially ventilator-associated pneumonia, are the preferential use of oro-tracheal rather than naso-tracheal tubes in patients who receive mechanically assisted ventilation, the use of noninvasive ventilation to reduce the need for and duration of endotracheal intubation, changing the breathing circuits of ventilators when they malfunction or are visibly contaminated, and (when feasible) the use of an endotracheal tube with a dorsal lumen to allow drainage of respiratory secretions; no recommendations were made about the use of sucralfate, histamine-2 receptor antagonists, or antacids for stress-bleeding prophylaxis. For prevention of health-care--associated Legionnaires disease, the changes include maintaining potable hot water at temperatures not suitable for amplification of Legionella spp., considering routine culturing of water samples from the potable water system of a facility's organ-transplant unit when it is done as part of the facility's comprehensive program to prevent and control health-care--associated Legionnaires disease, and initiating an investigation for the source of Legionella spp. when one definite or one possible case of laboratory-confirmed health-care--associated Legionnaires disease is identified in an inpatient hemopoietic stem-cell transplant (HSCT) recipient or in two or more HSCT recipients who had visited an outpatient HSCT unit during all or part of the 2-10 day period before illness onset. In the section on aspergillosis, the revised recommendations include the use of a room with high-efficiency particulate air filters rather than laminar airflow as the protective environment for allogeneic HSCT recipients and the use of high-efficiency respiratory-protection devices (e.g., N95 respirators) by severely immunocompromised patients when they leave their rooms when dust-generating activities are ongoing in the facility. In the respiratory syncytial virus (RSV) section, the new recommendation is to determine, on a case-by-case basis, whether to administer monoclonal antibody (palivizumab) to certain infants and children aged <24 months who were born prematurely and are at high risk for RSV infection. In the section on influenza, the new recommendations include the addition of oseltamivir (to amantadine and rimantadine) for prophylaxis of all patients without influenza illness and oseltamivir and zanamivir (to amantadine and rimantadine) as treatment for patients who are acutely ill with influenza in a unit where an influenza outbreak is recognized. In addition to the revised recommendations, the guideline contains new sections on pertussis and lower respiratory tract infections caused by adenovirus and human parainfluenza viruses and refers readers to the source of updated information about prevention and control of severe acute respiratory syndrome.


title: Localization and membrane topology of coronavirus nonstructural protein 4: involvement of the early secretory pathway in replication.

text: The coronavirus nonstructural proteins (nsp's) derived from the replicase polyproteins collectively constitute the viral replication complexes, which are anchored to double-membrane vesicles. Little is known about the biogenesis of these complexes, the membrane anchoring of which is probably mediated by nsp3, nsp4, and nsp6, as they contain several putative transmembrane domains. As a first step to getting more insight into the formation of the coronavirus replication complex, the membrane topology, processing, and subcellular localization of nsp4 of the mouse hepatitis virus (MHV) and severe acute respiratory syndrome-associated coronavirus (SARS-CoV) were elucidated in this study. Both nsp4 proteins became N glycosylated, while their amino and carboxy termini were localized to the cytoplasm. These observations imply nsp4 to assemble in the membrane as a tetraspanning transmembrane protein with a Nendo/Cendo topology. The amino terminus of SARS-CoV nsp4, but not that of MHV nsp4, was shown to be (partially) processed by signal peptidase. nsp4 localized to the endoplasmic reticulum (ER) when expressed alone but was recruited to the replication complexes in infected cells. nsp4 present in these complexes did not colocalize with markers of the ER or Golgi apparatus, while the susceptibility of its sugars to endoglycosidase H indicated that the protein had also not traveled trough the latter compartment. The important role of the early secretory pathway in formation of the replication complexes was also demonstrated by the inhibition of coronaviral replication when the ER export machinery was blocked by use of the kinase inhibitor H89 or by expression of a mutant, Sar1[H79G].


title: The formation and function of extracellular enveloped vaccinia virus.

text: Vaccinia virus produces four different types of virion from each infected cell called intracellular mature virus (IMV), intracellular enveloped virus (IEV), cell-associated enveloped virus (CEV) and extracellular enveloped virus (EEV). These virions have different abundance, structure, location and roles in the virus life-cycle. Here, the formation and function of these virions are considered with emphasis on the EEV form and its precursors, IEV and CEV. IMV is the most abundant form of virus and is retained in cells until lysis; it is a robust, stable virion and is well suited to transmit infection between hosts. IEV is formed by wrapping of IMV with intracellular membranes, and is an intermediate between IMV and CEV/EEV that enables efficient virus dissemination to the cell surface on microtubules. CEV induces the formation of actin tails that drive CEV particles away from the cell and is important for cell-to-cell spread. Lastly, EEV mediates the long-range dissemination of virus in cell culture and, probably, in vivo. Seven virus-encoded proteins have been identified that are components of IEV, and five of them are present in CEV or EEV. The roles of these proteins in virus morphogenesis and dissemination, and as targets for neutralizing antibody are reviewed. The production of several different virus particles in the VV replication cycle represents a coordinated strategy to exploit cell biology to promote virus spread and to aid virus evasion of antibody and complement.


title: Deformation of the Femoropopliteal Segment: Effect of Stent Length, Location, Flexibility, and Curvature.

text: PURPOSE To quantify the deformation behavior of the diseased femoropopliteal segment and assess the change to deformation behavior due to various stent placements. METHODS The length and curvature changes of 6 femoropopliteal segments (the right and left superficial femoral and popliteal arteries) from 3 cadavers were measured in 3-dimensional space based on rotational angiography image data in straight leg and flexed hip/knee (50°/90°) positions before and after placement of nitinol stents of varying type (EverFlex, Misago, and BioMimics 3D) and length (60, 100, and 200 mm) in different locations along the arteries. Three-dimensional centerline data were extracted for the measurements. RESULTS All 6 femoropopliteal cadaver segments displayed signs of peripheral artery disease. Hip/knee flexion resulted in vessel shortening and increases in the mean and maximum vessel curvatures in all cases. Location-specific results of the unstented arteries showed that magnitudes of vessel length and curvature change vary as a function of vessel length. The average shortening of the entire femoropopliteal segment due to flexion was observed at 10.7%±0.7%, which was reduced to 8.1%±0.9% after stent deployment. Average and maximum curvatures of the unstented segment increased due to flexion (average: 0.008±0.002 mm-1 to 0.019±0.006 mm-1, maximum: 0.030±0.009 mm-1 to 0.091±0.045 mm-1). After stent deployment, average and maximum curvatures of the flexed stented segments increased compared with the flexed unstented segments (average: 0.019±0.006 mm-1 to 0.022±0.004 mm-1, maximum: 0.091±0.045 mm-1 to 0.103±0.025 mm-1). The most flexurally stiff stent demonstrated the least ability to axially shorten during flexion of the leg at the knee joint. CONCLUSION The deformation characteristics of the femoropopliteal segment change in the presence of a stent, with the change to the deformation behavior dependent on stent type, stent length, location, flexibility, and intrinsic centerline curvature.


The coronaviruslike superfamily.


title: Evaluation of antibiotic-loaded calcium phosphate bone cement in an cranium-infected experimental model.

text: Treatment of calvarial defects has remained a challenge in reconstruction surgery, especially because of infection at these sites. We produced a bactericidal biomaterial for treating infected bone defects by using calcium phosphate bone cement mixed with antibiotics. We evaluated the usefulness of this material mixed with the antibiotic vancomycin in a cranium-infected rat model. The concentration of vancomycin used was 5.0 wt%, as reported in our previous study. In order to establish the rat model, a cranium defect (diameter, 5 mm) was made that was infected with methicillin-resistant Staphylococcus aureus (MRSA). Thirty-six rats were divided into 6 groups depending on whether an autologous graft or bone cement with or without antibiotic was used for the defect. After 1 and 4 weeks, abscess formation was checked, tissue bacterial counts were determined, and pathological examination was performed. At both 1 and 4 weeks, no MRSA was detected on tissue bacterial culture or pathological examination in groups that received bone cement with antibiotics. In groups that received bone cement without antibiotic, MRSA was detected, and the bone cement had compromised and disintegrated into several slices. In conclusion, bone cement that contains antibiotics appears to be effective not only for reconstruction in cases of cranial defect, but also in terms of preventing infection.


PETITION FOR A WRIT OF CERTIORARI.


title: Exacerbated viral hepatitis in IFN-gamma receptor-deficient mice is not suppressed by IL-12.

text: Both IL-12 and IFN-gamma have been implicated as principal inducers of type 1 immune responses required for the elimination of intracellular pathogens, such as viruses. We examined the in vivo antiviral role of both cytokines during coronavirus-induced hepatitis in a mouse hepatitis virus (MHV) model. The absence of IFN-gamma function in mice with a targeted disruption of the IFN-gamma R alpha-chain gene (IFN-gamma R -/-) resulted in increased susceptibility to coronaviral hepatitis associated with augmented viral replication and increased hepatocellular injury. The mutant mice showed a type 1 lymphokine response characterized by the normal high IFN-gamma and low IL-4 production. Unlike MHV-infected wild-type mice, however, the mutant IFN-gamma R -/- mice showed no increase in IL-12 p4O gene expression, similar to that in naive animals. IL-12 treatment failed to restore host resistance in IFN-gamma R -/- mice, but significantly protected MHV-susceptible C57BL/6 mice against lethal infection, although less than IFN-gamma treatment. Mice protected by IL-12 or IFN-gamma showed resistance against an otherwise lethal second MHV infection. Our data demonstrate that despite reduced IL-12 gene expression and defective IFN-gamma R function, virus-induced IFN-gamma production can occur. Furthermore, they emphasize the pivotal antiviral role of IFN-gamma in protection against acute coronavirus-induced hepatitis.


title: Human rhinovirus 3C protease: generation of pharmacophore models for peptidic and nonpeptidic inhibitors and their application in virtual screening.

text: Three-dimensional pharmacophore models for peptidic and small organic nonpeptidic inhibitors of the human rhinovirus 3C protease were generated in a structure-based as well as in a ligand-based approach, using the software package Catalyst. The inhibitors possess an electrophilic moiety, often a Michael acceptor function, which covalently binds to a cysteine in the active site of the enzyme. Since this process presents the key step for virus inactivation, the creation of a new function in Catalyst was required in order to include this decisive functionality into the pharmacophore models. In the present study we focus on this feature definition process because it presents an innovative strategy to expand the pharmacophore description ability of the Catalyst software to also include covalent bonds between ligand and binding site. The resulting hypotheses were then used for virtual screening of 3D databases in order to verify their quality and to search for structurally diverse, possible new lead substances.


title: Identification of contaminant sources in enclosed spaces by a single sensor.

text: UNLABELLED To protect occupants from infectious diseases or possible chemical/biological agents released by a terrorist in an enclosed space, such as an airliner cabin, it is critical to identify gaseous contaminant source locations and strengths. This paper identified the source locations and strengths by solving inverse contaminant transport with the quasi-reversibility (QR) and pseudo-reversibility (PR) methods. The QR method replaces the second-order diffusion term in the contaminant transport equation with a fourth-order stabilization term. By using the airflow pattern calculated by computational fluid dynamics (CFD) and the time when the peak contaminant concentration was measured by a sensor in downstream, the QR method solves the backward probability density function (PDF) of contaminant source location. The PR method reverses the airflow calculated by CFD and solves the PDF in the same manner as the QR method. The position with the highest PDF is the location of the contaminant source. The source strength can be further determined by scaling the nominal contaminant concentration computed by CFD with the concentration measured by the sensor. By using a two-dimensional and a three-dimensional aircraft cabin as examples of enclosed spaces, the two methods can identify contaminant source locations and strengths in the cabins if the sensors are placed in the downstream location of the sources. The QR method performed slightly better than the PR method but with a longer computing time. PRACTICAL IMPLICATIONS The paper presents a method that can be used to find a gaseous contaminant source location and determine its strength in enclosed spaces with the data of contaminant concentration measured by one sensor. The method can be a very useful tool to find where, what, and how the contamination has happened. The method is also useful for optimally placing sensors in enclosed spaces. The results can be applied to develop appropriate measures to protect occupants in enclosed environments from infectious diseases or chemical/biological warfare agents released by a terrorist.


title: Subgenomic RNA synthesis directed by a synthetic defective interfering RNA of mouse hepatitis virus: a study of coronavirus transcription initiation.

text: We have used a full-length cDNA clone of a mouse hepatitis virus strain A59 defective interfering (DI) RNA, pMIDI-C, and cassette mutagenesis to study the mechanism of coronavirus subgenomic mRNA synthesis. Promoter sequences closely resembling those of subgenomic mRNAs 3 and 7 were inserted into MIDI-C. Both subgenomic RNA promoters gave rise to the synthesis of a subgenomic DI RNA in virus-infected and DI RNA-transfected cells. From a mutagenic analysis of the promoters we concluded the following. (i) The extent of base pairing between the leader RNA and the intergenic promoter sequence does not control subgenomic RNA abundance. (ii) Promoter recognition does not rely on base pairing only. Presumably, transcription initiation requires recognition of the promoter sequence by the transcriptase. (iii) Fusion of leader and body sequences takes place at multiple--possibly random--sites within the intergenic promoter sequence. A model is presented in which, prior to elongation, the leader RNA is trimmed by a processive 3'-->5' nuclease.


title: Highlights from the 13th European Congress of Clinical Microbiology and Infectious Diseases in Glasgow, Scotland, May 10-13, 2003. The complex world of infectious diseases.

text: At the 13th European Congress of Clinical Microbiology and Infectious Diseases, held in Glasgow, Scotland, May 10-13, 2003, the latest developments in clinical microbiology and the treatment of infectious diseases were presented alongside recent progress on molecular aspects of diagnosis and emerging patterns of infection. Around 5,000 delegates from more than 80 countries attended the congress, which saw the presentation of more than 400 oral communications and 1,700 posters. In addition to a historical session looking at Scotland's own contribution to the control of infectious diseases, the meeting involved up to six parallel sessions a day, looking at all the major aspects of infectious diseases, treatment, surveillance, epidemiology and drug pharmacodynamics and pharmacokinetics. The organizers also organized a Late Breaker symposium on severe acute respiratory syndrome. The topics likely to be of most interest to Drug News and Perspectives readers are described here.


title: [Initial division of the left renal vein before dissection of left renal vein occluded by intracaval tumor thrombus].

text: Between November 2008 and March 2010, we performed initial division of the left renal vein occluded by the tumor thrombus in six cases of left renal cancer at Toranomon Hospital. The left renal vein was completely occluded by the tumor thrombus in all cases. In order to ligate the left renal artery first behind the dilated left renal vein, we must dissect the left kidney with arterial blood flow. Massive bleeding from the numerous engorged collateral veins around the left kidney is inevitable. Furthermore, access to the left renal artery is difficult because of the large tumor. We therefore initially divided the left renal vein without arterial blood flow followed by division of the left renal artery. After nephrectomy by dissecting the tumor without blood flow we extirpated the intracaval tumor thrombus. The median time of the operation was 7 hours 35 minutes and the median amount of blood loss was 2,869 ml. The tumor stage was pT3b in four cases and pT3c in two cases. No complications were observed during and after surgery except for one case of lymphocele and another case of chylous ascites. The initial division of the left renal vein is considered to be a useful surgical approach in left renal cancer with occluded left renal vein, especially when the tumor is large.


title: Mathematical modeling of subgenomic hepatitis C virus replication in Huh-7 cells.

text: Cell-based hepatitis C virus (HCV) replicon systems have provided a means for understanding HCV replication mechanisms and for testing new antiviral agents. We describe here a mathematical model of HCV replication that assumes that the translation of the HCV polyprotein occurs in the cytoplasm, that HCV RNA synthesis occurs in vesicular-membrane structures, and that the strategy of replication involves a double-stranded RNA intermediate. Our results shed light on the intracellular dynamics of subgenomic HCV RNA replication from transfection to steady state within Huh-7 cells. We predict the following: (i) about 6 x 10(3) ribosomes are involved in generating millions of HCV NS5B-polymerase molecules in a Huh-7 cell, (ii) the observed 10:1 asymmetry of plus- to minus-strand RNA levels can be explained by a higher-affinity (200-fold) interaction of HCV NS5B polymerase-containing replication complexes with HCV minus-strand RNA over HCV plus-strand RNA in order to initiate synthesis, (iii) the latter higher affinity can also account for the observed approximately 6:1 plus-strand/minus-strand ratio in vesicular-membrane structures, and (iv) the introduction of higher numbers of HCV plus-strand RNA by transfection leads to faster attainment of steady-state but does not change the steady-state HCV RNA level. Fully permissive HCV replication systems have been developed, and the model presented here is a first step toward building a comprehensive model for complete HCV replication. Moreover, the model can serve as an important tool in understanding HCV replication mechanisms and should prove useful in designing and evaluating new antivirals against HCV.


title: [Respiratory complications of sickle cell anemia in children: the acute chest syndrome].

text: The acute chest syndrome (ACS) is one of the most frequent complications of sickle cell disease. It affects mostly young children and counts for one quarter of mortality in the young sickle cell disease (SCD) population. This retrospective study evaluates the impact of ACS among hospitalizations for other complications of SCD in patients at the University Childrens' Hospital Reine Fabiola (Brussels, Belgium) in order to isolate clinical conditions associated with a high risk of ACS development. The medical records of all SCD patients aged up to 18 years admitted for all SCD related acute complications over a period of 13 month have been reviewed. Two patient groups have been formed based on the presence of an ACS within the study period. Epidemiologic data, medical history, the clinical presentation at admission but also blood counts in steady state, at admission and along the hospital stay were compared for a total of 96 hospital stays. There is no difference for age or hemoglobin phenotype between the two major patient groups. Male sex and having had a previous ACS episode in the past were significantly more important in the group of patients hospitalized for ACS. Thoracic pain in an SCD patient who doesn't show typical ACS symptoms should be interpreted as a risk factor for ACS. In conclusion, male sex, medical history of at least one ACS and thoracic pain at hospital admission are associated with high risk of developing ACS.


title: Cancer-specific viruses and the development of ONYX-015.

text: Viruses can be engineered to replicate in cancer cells selectively. This can be achieved using regulatory elements with tumor-selectivity, or using deletions of viral genes that are essential in normal cells but dispensable in tumor cells. The latter approach has been used to make viruses that depend on loss of RB or loss of p53. However, redundancy of function of viral proteins and cellular pathways has complicated this approach and required further viral engineering or a better understanding of cellular pathways to increase effectiveness. In spite of these complicating factors, ONYX-015, a prototype of this new class of therapeutic agents, has undergone extensive testing in the clinic, and has proven safe with promising signs of efficacy. Here, I summarize issues relating to selectivity of this agent and its clinical potential. Based on clinical data and new basic discoveries, several approaches are suggested to improve the efficacy of this agent in the clinic.


Prepare for Post-COVID-Pandemic Polio Problems.


Use of chaperones by general practitioners.


Evidence against rapid emergence of praziquantel resistance in Schistosoma haematobium, Kenya.


title: Sensitive and specific monoclonal antibody-based capture enzyme immunoassay for detection of nucleocapsid antigen in sera from patients with severe acute respiratory syndrome.

text: A rapid antigen test for the diagnosis of severe acute respiratory syndrome (SARS) is essential for control of this disease at the point of management. The nucleocapsid (N) protein of SARS-associated coronavirus (SARS-CoV) is abundantly expressed in infected-cell culture filtrate as demonstrable by Western blotting using convalescent-phase sera from patients with SARS. We used monoclonal antibodies specifically directed against N protein to establish a sensitive antigen capture sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of SARS-CoV. The assay employed a mixture of three monoclonal antibodies for capture and rabbit polyclonal antibodies for detection of serum antigen in 32 cases of clinically probable SARS as defined by the World Health Organization during the epidemic in Guangzhou, China. Recombinant N protein was used as a standard to establish a detection sensitivity of approximated 50 pg/ml. The linear range of detection in clinical specimens was from 100 pg/ml to 3.2 ng/ml. Using a panel of sera collected at different points in time, the amount of circulating N antigen was found to peak 6 to 10 days after the onset of symptoms. The sensitivity of the assay was 84.6% in 13 serologically confirmed SARS patients with blood taken during the first 10 days after the onset of symptoms (11 of 13). The specificity of the assay was 98.5% in 1,272 healthy individuals (1,253 of 1,272). There was no cross-reaction with other human and animal coronaviruses in this assay. In conclusion, a sensitive and quantitative antigen capture ELISA was established for the early diagnosis and disease monitoring of SARS-CoV infection.


title: First experimental evidence of recombination in infectious bronchitis virus. Recombination in IBV.

text: A high frequency of recombination has been shown to occur during replication of the coronavirus mouse hepatitis virus (MHV) in vitro as well as in vivo. Although sequencing of field strains of coronavirus infectious bronchitis virus (IBV) has indicated that IBV strains also undergo recombination, there has been no experimental evidence to support this deduction. To investigate whether recombination occurs in IBV, embryonated eggs were coinfected with IBV-Beaudette and IBV-M41. Potential recombinants were detected by strain-specific polymerase chain reaction (PCR) amplifications, using oligonucleotides corresponding to regions in the 3' end of the genome. Sequencing of the PCR products confirmed that a number of recombinations had occurred between the two strains.


title: Early evolution and the origin of eukaryotes.

text: Our understanding of evolutionary relationships in the eukaryotic world has been revolutionized by molecular systematics. Phylogenies based upon comparisons of rRNAs define five major eukaryotic assemblages plus a series of paraphyletic protist lineages. Comparison of conserved genes that were duplicated prior to the divergence of eubacteria, archaebacteria, and eukaryotes, positions the root of the universal tree within the eubacterial line of descent. In this review a novel model is presented which uses the rRNA and protein based phylogenies to describe the evolutionary origins of eukaryotes.


Neurovirulence for mice of soluble receptor-resistant mutants of murine coronavirus JHMV.


title: Crystal structure of the severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein dimerization domain reveals evolutionary linkage between corona- and arteriviridae.

text: The causative agent of severe acute respiratory syndrome (SARS) is the SARS-associated coronavirus, SARS-CoV. The nucleocapsid (N) protein plays an essential role in SARS-CoV genome packaging and virion assembly. We have previously shown that SARS-CoV N protein forms a dimer in solution through its C-terminal domain. In this study, the crystal structure of the dimerization domain, consisting of residues 270-370, is determined to 1.75A resolution. The structure shows a dimer with extensive interactions between the two subunits, suggesting that the dimeric form of the N protein is the functional unit in vivo. Although lacking significant sequence similarity, the dimerization domain of SARS-CoV N protein has a fold similar to that of the nucleocapsid protein of the porcine reproductive and respiratory syndrome virus. This finding provides structural evidence of the evolutionary link between Coronaviridae and Arteriviridae, suggesting that the N proteins of both viruses have a common origin.


title: Swollen-head syndrome in broiler chickens.

text: Swollen-head syndrome is a disease seen in broiler chickens between 4 and 6 weeks of age in Southern Africa. It appears to be caused by a mixed coronavirus and Escherichia coli infection. The coronavirus appears to be of a hitherto unrecorded serotype. The disease is controlled by an attenuated live-virus vaccine and antibacterial medication.


title: A Qualitative Evaluation of Language Models on Automatic Question-Answering for COVID-19.

text: COVID-19 has resulted in an ongoing pandemic and as of 12 June 2020, has caused more than 7.4 million cases and over 418,000 deaths. The highly dynamic and rapidly evolving situation with COVID-19 has made it difficult to access accurate, on-demand information regarding the disease. Online communities, forums, and social media provide potential venues to search for relevant questions and answers, or post questions and seek answers from other members. However, due to the nature of such sites, there are always a limited number of relevant questions and responses to search from, and posted questions are rarely answered immediately. With the advancements in the field of natural language processing, particularly in the domain of language models, it has become possible to design chatbots that can automatically answer consumer questions. However, such models are rarely applied and evaluated in the healthcare domain, to meet the information needs with accurate and up-to-date healthcare data. In this paper, we propose to apply a language model for automatically answering questions related to COVID-19 and qualitatively evaluate the generated responses. We utilized the GPT-2 language model and applied transfer learning to retrain it on the COVID-19 Open Research Dataset (CORD-19) corpus. In order to improve the quality of the generated responses, we applied 4 different approaches, namely tf-idf, BERT, BioBERT, and USE to filter and retain relevant sentences in the responses. In the performance evaluation step, we asked two medical experts to rate the responses. We found that BERT and BioBERT, on average, outperform both tf-idf and USE in relevance-based sentence filtering tasks. Additionally, based on the chatbot, we created a user-friendly interactive web application to be hosted online.


title: Why examining the desirability of health technology matters.

text: Although technology is ubiquitous in healthcare, its impact on people's perceptions and lives is poorly understood. Fresh insights are required to meet current and future technology-related policy challenges. Keeping a population healthy requires considering not only technologies that are used in clinical settings (diagnostic, therapeutic, palliative), but also those used in the community (home care, self-care, technical aids) and those that affect health more broadly (health promotion technologies, occupational health technologies). At the policy making level, understanding the desirability of health technology may prove to be more important than simply appraising its affordability.


The acute respiratory distress syndrome.


title: Progress in computational approach to drug development against SARS.

text: Since the outbreak of SARS (severe acute respiratory syndrome) in November 2002 in Southern China's Guangdong Province, considerable progress has been made in the development of drugs for SARS therapy. The present mini review is focused on the area of computer-aided drug discovery, i.e., the advances achieved mainly from the approaches of structural bioinformatics, pharmacophore modeling, molecular docking, peptide-cleavage site prediction, and other computational means. It is highlighted that the compounds C(28)H(34)O(4)N(7)Cl, C(21)H(36(O)5)N(6) and C(21)H(36)O(5)N(6) (Wei et al., Amino Acids, 2006, 31: 73-80), as well as KZ7088 (Chou et al. Biochem. Biophys. Res. Commun., 2003, 308: 148-151), a derivative of AG7088, might be the promising candidates for further investigation, and that the octapeptides ATLQAIAS and ATLQAENV, as well as AVLQSGFR, might be converted to effective inhibitors against the SARS enzyme. Meanwhile, how to modify these octapeptides based on the "distorted key" theory to make them become potent inhibitors is explicitly elucidated. Finally, a brief introduction is given for how to use computer-generated graphs to rapidly diagnose SARS coronavirus.


title: Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot.

text: From Babesia caballi in vitro cultures a preparation of 100% infected erythrocytes was obtained. From this, B. caballi antigens were extracted with the detergent 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate (CHAPS) and used as ELISA antigens. A control antigen of normal erythrocytes from the same donor horse was prepared in an identical manner. The ELISA and Western blot were validated by testing of sera from horses experimentally infected with B. caballi or B. equi or not infected with Babesia spp. ELISA and Western blot results were compared with those obtained by the immunofluorescence antibody test (IFAT) and complement fixation test (CFT). The sensitivity of the ELISA of 98.3% obtained for sera from day 14 after infection was superior to the Western blot (94.9%), the IFAT (96.6%) and the CFT (28.8%). No positive results were obtained in the ELISA and Western blot with 106 sera from horses not infected with Babesia spp. resulting in a calculated specificity of 100% for both tests. Cross reactions of B. equi-positive sera did occur to a larger extent in the ELISA (20%) than in the IFAT (4%). No cross reactions were observed with the Western blot and the CFT. The higher sensitivity of the ELISA was also demonstrated by testing of 132 field sera: more positive results were obtained by ELISA (112) as compared to IFAT (92) or CFT (41). The validity of these results was confirmed by testing of sera by Western blot. The ELISA as the most sensitive test provides the best method for the identification of carrier horses to prevent the introduction into non-endemic areas (export testing).(ABSTRACT TRUNCATED AT 250 WORDS)


title: The effect of antioxidant supplementation on bacterial translocation after intestinal ischemia and reperfusion.

text: The intestine is highly sensitive to ischemia/reperfusion (I/R) injury. Intestinal I/R may cause local tissue injury and disruption of the intestinal mucosal barrier, allowing the passage of viable bacteria and endotoxins from the gastrointestinal lumen to distant organs. This phenomenon, known as bacterial translocation (BT), may lead to systemic disorders with high morbidity and mortality. Oxidative stress mediators such as reactive oxygen species, polymorphonuclear neutrophils and nitric oxide are believed to contribute to the intestinal I/R injury. Many antioxidants have shown protective effects against I/R injury of various organs. The present article provides an overview of studies investigating the effect of antioxidant supplementation on BT after intestinal I/R.


Vet nurse consultations could help financial recovery from Covid-19.


title: Enterotropic mouse hepatitis virus.

text: Mouse hepatitis virus [MHV], the coronavirus of the mouse, is the most common viral pathogen in contemporary laboratory mouse colonies throughout the world. It is highly contagious with variable clinical manifestations. The majority of infections are subclinical, but can still significantly influence biological responses, thus interfering with research, mainly in the field of immunology. MHV has been intensively studied from a number of research perspectives and has become the prototype for studying the molecular biology of coronaviruses. MHV contains a single-stranded, positive-sense RNA genome ranging from 27 to 31 kb, which is divided into seven genes. Virions consist of four to five structural proteins. There are many MHV strains that vary in virulence, organotropism and cell tropism, and are constantly evolving by naturally occurring mutation and recombination. Based on pathogenesis studies MHV strains are usually grouped according to their primary tissue tropism into two biotypes: polytropic and enterotropic. Enterotropic strains of MHV replicate in the intestinal mucosa and only rarely spread to other tissues. No morphological structure of the virion has as yet been identified that is responsible for enterotropism. The course of an MHV infection is dependent on the virus strain and host factors. Generally, MHV causes an acute, self-limiting infection which is inapparent in adult mice. Neonates are highly susceptible to disease and show high mortality. In an enzootically infected colony, however, they are protected by maternally derived passive immunity. Detection of MHV infections depends on serological screening of colonies. MHV is controlled by culling and rederivation of the affected colony using hysterectomy or embryo transfer or by elimination through cessation of breeding.


title: Laparoscopic right colectomy with transvaginal extraction in a patient with prior pancreaticoduodenectomy.

text: INTRODUCTION Previous complicated abdominal surgeries such as pancreaticoduodenectomy with large abdominal incisions may keep the surgeons away from major laparoscopic procedures. To the best of our knowledge, there is no published study that shows the feasibility of major laparoscopic surgery in a patient with previous pancreaticoduodenectomy. CASE REPORT A 68-year-old female (BMI 27 kg/m2, ASA II), was admitted for anemia. Her medical history included an open pancreaticoduodenectomy four years ago for chronic pancreatitis. She had an abdominal Mercedes incision. Computed tomography and colonoscopy showed a 5-cm cecal mass with a histological diagnosis of adenocarcinoma. We performed a totally laparoscopic right hemicolectomy and intracorporeal ileotransverse anastomosis. The specimen was extracted through the vagina. The operating time was 500 minutes and the blood loss was 400 ml. The patient was uneventfully discharged on postoperative day four. CONCLUSIONS Laparoscopic colon surgery can be feasible and safe despite previous extensive abdominal surgeries such as pancreaticoduodenectomy. Moreover, laparoscopic surgery in these cases can also be completed with intracorporeal anastomosis and specimen extraction through a natural orifice.


title: Gene 5 of the avian coronavirus infectious bronchitis virus is not essential for replication.

text: The avian coronavirus Infectious bronchitis virus (IBV), like other coronaviruses, expresses several small nonstructural (ns) proteins in addition to those from gene 1 (replicase) and the structural proteins. These coronavirus ns genes differ both in number and in amino acid similarity between the coronavirus groups but show some concordance within a group or subgroup. The functions and requirements of the small ns gene products remain to be elucidated. With the advent of reverse genetics for coronaviruses, the first steps in elucidating their role can be investigated. We have used our reverse genetics system for IBV (R. Casais, V. Thiel, S. G. Siddell, D. Cavanagh, and P. Britton, J. Virol. 75:12359-12369, 2001) to investigate the requirement of IBV gene 5 for replication in vivo, in ovo, and ex vivo. We produced a series of recombinant viruses, with an isogenic background, in which complete expression of gene 5 products was prevented by the inactivation of gene 5 following scrambling of the transcription-associated sequence, thereby preventing the expression of IBV subgenomic mRNA 5, or scrambling either separately or together of the translation initiation codons for the two gene 5 products. As all of the recombinant viruses replicated very similarly to the wild-type virus, Beau-R, we conclude that the IBV gene 5 products are not essential for IBV replication per se and that they are accessory proteins.


title: Aspirin combined with mechanical measures to prevent venous thromboembolism after total knee arthroplasty: a randomized controlled trial.

text: BACKGROUND Venous thromboembolism (VTE) is an important complication after major orthopedic surgery. Pharmaceutical methods represent the main strategy of VTE prevention. The use of aspirin in VTE prevention is still controversial worldwide, especially in China. The purpose of this study was to evaluate the role of aspirin combined with mechanical measures in the prevention of VTE after total knee arthroplasty (TKA). METHODS Between January 2012 and May 2013 and in accordance with the inclusion criteria, 120 patients undergoing TKA were randomly allocated to two groups. To prevent VTE, patients in group A received aspirin combined with mechanical measures postoperatively, while patients in group B received low-molecular-weight heparin (LMWH) sodium and rivaroxaban sequentially in combination with mechanical measures postoperatively. All surgeries were performed by one surgeon using a posterior-stabilized cemented prosthesis. The two groups were followed up and compared for the incidence of deep vein thrombosis (DVT) by duplex ultrasound scan and clinical VTE events. The adverse events, the blood loss index, and the cost of VTE prevention were also compared. RESULTS DVT was detected in 10 of 60 patients in group A (16.7%, 95% CI: 7.3%-26.1%) compared with 11 of 60 in group B (18.3%, 95% CI: 8.5%-27.8%) (P = 0.500). There is no statistical evidence supporting the inferior effect of aspirin in preventing DVT as compared with the other medications. There were no cases of symptomatic VTE or death during the follow-up period. Area of ecchymosis was lower in group A than in group B, and the differences were statistically significant. Patients in group A had the lower blood loss index as compared with patients in group B. No transfusion cases were found in both groups. The differences were statistically significant. The cost of VTE prevention analysis indicated a cost reduction using aspirin in group A compared with using LMWH and rivaroxaban in group B. CONCLUSION Aspirin combined with mechanical measures had a good effect on prevention of VTE after TKA and resulted in lower cost, less blood loss, and less subcutaneous ecchymosis.


Lessons from the SARS crisis--more relevant than ever.


title: Two genotypes of canine coronavirus simultaneously detected in the fecal samples of dogs with diarrhea.

text: Sixty-nine fecal samples from diarrheic puppies were examined by reverse transcription-PCR assays for the M and the S genes of canine coronaviruses (CCoVs). The isolates in 10 samples were recognized as CCoV type I, and the isolates in 6 samples were recognized as CCoV type II, while isolates of both genotypes were simultaneously detected in 53 samples.


title: Detection of influenza antigen with rapid antibody-based tests after intranasal influenza vaccination (FluMist).

text: Rapid tests for influenza antigen detection are frequently used, but it is not known how receipt of intranasal influenza vaccine affects results of these tests. We tested healthy adults who received either intranasal or intramuscular influenza vaccine. Of the 14 intranasal vaccine recipients, 7 (50%) had a direct fluorescent antibody test (DFA) result and 2 (14%) had an enzyme immunoassay (EIA) result that was positive for influenza antigen within 7 days after vaccination. No subjects had positive EIA results on day 12 or 13 after vaccination. For some intranasal vaccine recipients, rapid influenza-antigen detection tests yield positive results within 1 week after vaccination.


HIV vaccine research a shot in the arm for immunologists.


title: Ultrastructural telepathology--remote EM-diagnostic via Internet.

text: New collaboratory opportunities in ultrastructural research and diagnostics are now available on the Internet through the combination of digital image acquisition, remote operation of modern digitally controlled and automated electron microscopes, and the development of software specifically tailored for collaboratory needs. Remote experts can examine samples directly, and unique instruments can be utilized from anywhere. In the case of diagnostic dilemmas, the second-opinion expert is no longer constrained by problems inherent in the interpretation of preselected images. The remote examiner can independently choose the area of interest on the sample as well as select the appropriate magnification for an accurate diagnosis. With these capabilities and together with teleconferencing tools and securely accessible databases on-line, telepathology can provide increased effectiveness and support for diagnostics, research, and teaching in many areas. The authors report their experience with remote electron microscope diagnoses of pathological samples using two different dynamic imaging systems and discuss the main technical issues encountered. It appears that only minor technical issues need to be resolved before ultrastructural telepathology can be promoted for routine use in areas with high-speed Internet access.


NIH response to open letter.


title: Infection of AtT20 murine pituitary tumour cells by mouse hepatitis virus strain A59: virus budding is restricted to the Golgi region.

text: AtT20 cells, a line of murine pituitary tumour cells that secrete adrenocorticotropic hormone (ACTH), have been infected with the coronavirus mouse hepatitis virus strain A59 (MHV-A59). Between 5% and 10% of AtT20 cells are susceptible to the infection. Unlike infections of fibroblastic sac- and 17Cl 1 cells, the infection of AtT20 cells does not lead to cell fusion, despite the production of the fusogenic E2 viral spike glycoprotein. Within infected AtT20 cells the second viral envelope glycoprotein, E1, is located in a perinuclear region; at least until very late in the infection it fails to accumulate to detectable levels in the rough endoplasmic reticulum (RER). By contrast to infection of sac- and 17Cl 1 cells, where the RER is a major site of assembly of progeny virions, in AtT20 cells budding of progeny virions is restricted to the Golgi cisternae, which eventually vesiculate, and peri-Golgi smooth membraned vesicles. Apparently, therefore, the intracellular compartments into which wild-type MHV-A59 buds are determined not by the virus but by the host cells. MHV-A59 infected cultures of AtT20 cells can be serially passaged without loss of the infection or increase in the proportion of infected cells; they become persistently infected carrier cultures. The progeny virus from serially passaged, infected AtT20 cells is apparently wild-type. It infects sac- cells and induces them to form syncitia. Within the sac- syncitia the viral E1 glycoprotein accumulates in the RER and many virions assemble there.


title: Biomarker variation in patients successfully treated with tocilizumab for severe coronavirus disease 2019 (COVID-19): results of a multidisciplinary collaboration.

text: OBJECTIVES Acute respiratory distress syndrome (ARDS) related to SARS-CoV-2 is likely due to a cytokine storm characterised by a major release of pro-inflammatory cytokines, including interleukin-6 (IL-6). Blocking excessive IL-6 production might be the key to the COVID-19-ARDS treatment. Beneficial effects of IL-6 blockade using a humanised anti-IL-6 receptor antibody, tocilizumab (TCZ) were previously reported in patients with COVID-19 related ARDS. The aim of the study was to study the variation over time of several biomarkers, demonstrated to be predictors of poor prognostic, in subjects successfully treated with TCZ for severe COVID-19. METHODS Retrospective analysis of a case series of patients with COVID-19-ARDS, evidenced by RT-PCR and lung CT-scan. Patients with increasing O2 requirements, within the window of opportunity for TCZ treatment (Day 7 to Day 17 after onset of symptoms) were treated with TCZ (2 infusions of 8 mg/kg). Demographic, biological and clinical data were collected from the patients' files. Serum levels of CRP, ferritin, fibrinogen, lymphocytes, platelets, creatinine, D-dimer and liver enzymes were assayed at the time of the first TCZ administration, then every two days for 8 days. RESULTS 40 patients were treated with TCZ. Most of them had several comorbidities, and all had multiple biological abnormalities (lymphopenia, increased CRP, ferritin, fibrinogen, D-dimer, liver enzymes). 30 patients (75%) benefited from TCZ and 10 patients died. In the survivors, following TCZ administration CRP decreased dramatically as early as day 4 (-86.7%, p<0.0001) and returned to normal at day 6. Fibrinogen and lymphocyte count returned to normal values at day 6. Ferritin also decreased significantly. No significant change was observed for D-dimer (p=0.68) and other studied biomarkers (haemoglobin, leucocyte count, AST). CONCLUSIONS In patients with COVID-19 acute respiratory distress syndrome, treatment with TCZ resulted in favourable evolution in 75% of the cases. Biomarkers of inflammation (CRP, ferritin, fibrinogen) decreased dramatically as early as the 4th day after TCZ injection. Lymphopenia, a predictor of poor prognostic, was reversed 6 days after TCZ injection.


title: Antiidiotypic vaccination against murine coronavirus infection.

text: Rabbit polyclonal antiidiotypic antibodies were generated against a neutralizing mAb specific for a conformational epitope on the S glycoprotein of murine hepatitis virus, strain A59 (MHV-A59). These anti-Id were directed predominantly against an Id that was undetectable in rabbit and rat anti-MHV-A59 sera and weakly represented in syngeneic and allogeneic antiviral sera. However, some partial idiotypic sharing was observed between the Id-bearing antibody and a mAb with a similar antigenic site specificity. The anti-Id inhibited the virus-binding and neutralizing activities of the immunizing antibody, demonstrating that they recognize paratope-associated idiotopes. Mice immunized with affinity-purified anti-Id developed MHV-A59-specific antibodies that neutralized viral infectivity to high titers. Moreover, these animals survived an otherwise lethal challenge with viral murine hepatitis virus, unlike control mice immunized with normal rabbit Ig. These results indicate that at least a subpopulation of the polyclonal anti-Id could induce a protective immune response directed toward a biologically important MHV-A59 epitope, and demonstrate the feasibility of antiidiotypic vaccination against a coronavirus infection.


ABC of chronic obstructive pulmonary disease. Acute exacerbations.


title: FELASA recommendations for the health monitoring of mouse, rat, hamster, guinea pig and rabbit colonies in breeding and experimental units.

text: The microbiological quality of experimental animals can critically influence animal welfare and the validity and reproducibility of research data. It is therefore important for breeding and experimental facilities to establish a laboratory animal health monitoring (HM) programme as an integrated part of any quality assurance system. FELASA has published recommendations for the HM of rodent and rabbit colonies in breeding and experimental units (Nicklas et al. Laboratory Animals, 2002), with the intention of harmonizing HM programmes. As stated in the preamble, these recommendations need to be adapted periodically to meet current developments in laboratory animal medicine. Accordingly, previous recommendations have been revised and shall be replaced by the present recommendations. These recommendations are aimed at all breeders and users of laboratory mice, rats, Syrian hamsters, guinea pigs and rabbits as well as diagnostic laboratories. They describe essential aspects of HM, such as the choice of agents, selection of animals and tissues for testing, frequency of sampling, commonly used test methods, interpretation of results and HM reporting. Compared with previous recommendations, more emphasis is put on the role of a person with sufficient understanding of the principles of HM, opportunistic agents, the use of sentinel animals (particularly under conditions of cage-level containment) and the interpretation and reporting of HM results. Relevant agents, testing frequencies and literature references are updated. Supplementary information on specific agents and the number of animals to be monitored and an example of a HM programme description is provided in the appendices.


title: Non-small cell lung cancer, version 2.2013.

text: These NCCN Guidelines Insights focus on the diagnostic evaluation of suspected lung cancer. This topic was the subject of a major update in the 2013 NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines) for Non-Small Cell Lung Cancer. The NCCN Guidelines Insights focus on the major updates in the NCCN Guidelines and discuss the new updates in greater detail.


Companion animals as reservoirs of eaeA+ Escherichia coli.


title: Preparedness for SARS in the UK in 2003.

text: Severe acute respiratory syndrome (SARS) has been described as the first major emerging infectious disease of the twenty-first century. Having initially emerged, almost unnoticed, in southern China, it rapidly spread across the globe. It severely tested national public health and health systems. However, it also resulted in rapid, intensive international collaboration, led by the World Health Organization, to elucidate its characteristics and cause and to contain its spread. The UK mounted a vigorous public health response. Some particular issues concerned: the practicalities of implementing exit screening had this been required; the likely efficacy of this and other control measures; the legal base for public health action; and the surge capacity in all systems should the disease have taken hold in the UK. We have used this experience of 2003 to inform our preparation of a framework for an integrated, escalating response to a future re-emergence of SARS according to the levels of disease activity worldwide. Recent cases confirm that SARS has not "gone away". We cannot be complacent about our contingency planning.


title: Laparoscopic esophagogastric circular stapled anastomosis: a modified technique to protect the esophagus.

text: Laparoscopic surgery is increasingly being applied to gastric cancer surgery, including proximal gastrectomy for the resection of cancer located in the upper gastric body. Despite the ease of use of stapling devices for end-to-end anastomosis, esophagogastric anastomosis is complicated by the narrow laparoscopic space, making the placement of an esophageal purse-string suture and anvil insertion into the fragile and contracted esophagus difficult. The aim of this study was to employ a novel esophagogastric anastomosis technique for laparoscopic surgery which may avoid esophageal breakdown. Eleven patients with early gastric cancer within the upper gastric body underwent laparoscopic proximal gastrectomy. The anvil of the stapler was introduced into the esophagus through a small gastrostomy, before transection of the esophagus. The esophageal-to-anterior gastric wall anastomosis was performed using a double-stapling technique, without the need to apply a purse-string suture. The mean operation time was 237 +/- 15 min and estimated blood loss was 39 +/- 21 ml. The postoperative course was uneventful in all 11 patients, with no anastomotic leakage observed. Two patients needed endoscopic balloon dilation of an anastomotic stricture 24 to 28 days postoperatively. This modified procedure of laparoscopic esophagogastric anastomosis after proximal gastrectomy for the resection of cancer is a simple, rapid, and atraumatic technique which reduces the risk of anastomotic insufficiency.


title: Identification of a novel coronavirus possibly associated with acute respiratory syndrome in alpacas (Vicugna pacos) in California, 2007.

text: Alpaca respiratory syndrome (ARS) was first recognized in California in October 2007. This syndrome is characterized by acute respiratory signs, high fever, and occasional sudden death, and has mostly been observed in pregnant alpacas (Vicugna pacos), although all signalments have been affected. A similarity in clinical signs to cases located on the East Coast of the United States was observed; however, a causative agent had not been identified. Preliminary diagnostic submissions to the California Animal Health and Food Safety Laboratory System (CAHFS) were negative for known bacterial, parasitic, fungal, and viral pathogens, as well as for toxins, making the etiology of this disease unknown. However, based on pathologic findings, a viral or toxic etiology was strongly considered. A novel coronavirus was recovered from lung tissue of a clinical case submitted to CAHFS. The coronavirus identity was confirmed in tissue culture by transmission electron microscopy and by sequence analysis of a conserved region within the viral genome. Statistical analysis calculating a serologic association between the serum virus neutralization antibody titer and coronavirus, the presence of exposure history on 40 animals with a history of ARS, and 167 controls provided an odds ratio of 121 (95% confidence interval: 36.54 and 402.84; P < 0.0001). The findings indicate that the ARS-associated coronavirus described is distinct from the previously reported gastrointestinal-associated coronavirus identified in alpaca herds.


title: Primary bacterial septic peritonitis in cats: 13 cases.

text: The purpose of this paper is to describe the signalment, clinical signs, laboratory results, culture results, and response to treatment for primary septic peritonitis in cats. This is a retrospective study of 12 client-owned animals. Medical records were reviewed for clinical findings, laboratory results, microbial culture results, radiographic findings, diagnosis, treatment, and outcome. The overall mortality rate for this group of cats was 31%, consistent with previous reports of septic peritonitis in cats. All cats that were both bradycardic and hypothermic on presentation did not survive. Other clinicopathological findings were consistent with previously reported cases of septic peritonitis in cats. Results suggest that clinicopathological findings and outcomes in cats with primary septic peritonitis are similar to those in cats with septic peritonitis from a determined cause. A specific mechanism of inoculation has yet to be determined, but an oral source of bacteria is suggested for cats with primary bacterial septic peritonitis.


Fine specificity and genetic restriction of T cell clones specific for mouse hepatitis virus, strain JHM.


title: Host protein interactions with the 3' end of bovine coronavirus RNA and the requirement of the poly(A) tail for coronavirus defective genome replication.

text: RNA viruses have 5' and 3' untranslated regions (UTRs) that contain specific signals for RNA synthesis. The coronavirus genome is capped at the 5' end and has a 3' UTR that consists of 300 to 500 nucleotides (nt) plus a poly(A) tail. To further our understanding of coronavirus replication, we have begun to examine the involvement of host factors in this process for two group II viruses, bovine coronavirus (BCV) and mouse hepatitis coronavirus (MHV). Specific host protein interactions with the BCV 3' UTR [287 nt plus poly(A) tail] were identified using gel mobility shift assays. Competition with the MHV 3' UTR [301 nt plus poly(A) tail] suggests that the interactions are conserved for the two viruses. Proteins with molecular masses of 99, 95, and 73 kDa were detected in UV cross-linking experiments. Less heavily labeled proteins were also detected in the ranges of 40 to 50 and 30 kDa. The poly(A) tail was required for binding of the 73-kDa protein. Immunoprecipitation of UV-cross-linked proteins identified the 73-kDa protein as the cytoplasmic poly(A)-binding protein (PABP). Replication of the defective genomes BCV Drep and MHV MIDI-C, along with several mutants, was used to determine the importance of the poly(A) tail. Defective genomes with shortened poly(A) tails consisting of 5 or 10 A residues were replicated after transfection into helper virus-infected cells. BCV Drep RNA that lacked a poly(A) tail did not replicate, whereas replication of MHV MIDI-C RNA with a deleted tail was detected after several virus passages. All mutants exhibited delayed kinetics of replication. Detectable extension or addition of the poly(A) tail to the mutants correlated with the appearance of these RNAs in the replication assay. RNAs with shortened poly(A) tails exhibited less in vitro PABP binding, suggesting that decreased interactions with the protein may affect RNA replication. The data strongly indicate that the poly(A) tail is an important cis-acting signal for coronavirus replication.


Rigors.


Sudden infant death syndrome.


title: Evaluation of a quantitative real-time PCR for the detection of respiratory syncytial virus in pulmonary diseases.

text: Respiratory syncytial virus (RSV) is known to cause acute lower respiratory tract infections (ARI) in young children and is involved in exacerbation of chronic obstructive pulmonary disease (COPD) in adults. The role of RSV in stable COPD and the viral load in different respiratory diseases has not been investigated to date. The present authors established and evaluated a quantitative TaqMan real-time polymerase chain reaction assay specific for RSV subgroup A. Absolute quantification for the determination of viral load input was achieved using a control plasmid. The assay allowed for a quantification over a >6-log range and a detection limit of <10 RSV copies per reaction mixture. The assay was 30 times more sensitive than conventional nested polymerase chain reaction assays. Interassay SD was 1.3 and coefficient of variation 4.7% on average. Clinical specimens from infants with ARI (n=62) and elderly adults with COPD (n=125) were compared for viral loads. A total of 47% RSV-positive samples were found in the ARI study group and 28% in the COPD study group. The viral load of both study groups was found to differ significantly. In the ARI study group the viral load was increased almost 2000-fold, suggesting acute infection in this group and former or latent infection in the COPD group. Respiratory syncytial virus-A specific TaqMan real-time polymerase chain reaction assay is a sensitive and rapid method for the determination of viral load in clinical samples. It enables differential statements concerning the involvement of respiratory syncytial virus in acute lower respiratory tract infections and chronic obstructive pulmonary disease to be achieved.


title: Severe acute respiratory syndrome coronavirus infection of mice transgenic for the human Angiotensin-converting enzyme 2 virus receptor.

text: Animal models for severe acute respiratory syndrome (SARS) coronavirus infection of humans are needed to elucidate SARS pathogenesis and develop vaccines and antivirals. We developed transgenic mice expressing human angiotensin-converting enzyme 2, a functional receptor for the virus, under the regulation of a global promoter. A transgenic lineage, designated AC70, was among the best characterized against SARS coronavirus infection, showing weight loss and other clinical manifestations before reaching 100% mortality within 8 days after intranasal infection. High virus titers were detected in the lungs and brains of transgene-positive (Tg+) mice on days 1 and 3 after infection. Inflammatory mediators were also detected in these tissues, coinciding with high levels of virus replication. Lower virus titers were also detected in other tissues, including blood. In contrast, infected transgene-negative (Tg-) mice survived without showing any clinical illness. Pathologic examination suggests that the extensive involvement of the central nervous system likely contributed to the death of Tg+ mice, even though viral pneumonia was present. Preliminary studies with mice of a second lineage, AC63, in which the transgene expression was considerably less abundant than that in the AC70 line, revealed that virus replication was largely restricted to the lungs but not the brain. Importantly, despite significant weight loss, infected Tg+ AC63 mice eventually recovered from the illness without any mortality. The severity of the disease that developed in these transgenic mice--AC70 in particular--makes these mouse models valuable not only for evaluating the efficacy of antivirals and vaccines, but also for studying SARS coronavirus pathogenesis.


title: Elevated serum concentrations of inflammatory cytokines and chemokines in patients with haemorrhagic fever with renal syndrome.

text: OBJECTIVE Prospective case-control study, undertaken to investigate serum cytokine and chemokine concentrations during all clinical phases and in different clinical types of haemorrhagic fever with renal syndrome (HFRS). METHODS Serum was collected at various disease phases from patients with HFRS (n = 35) and healthy control subjects (n = 10). Tumour necrosis factor (TNF)-α, interleukin (IL)-6, IL-4, interferon (IFN)-γ, IL-8, interferon inducible protein-10 (IP-10) and chemokine (C-C motif) ligand 5 (also known as 'regulated upon activation, normal T-cell expressed and secreted' [RANTES]) were quantified using commercial enzyme-linked immunosorbent assay kits. RESULTS Serum concentrations of TNF-α, IL-6, IFN-γ, IL-8, IP-10 and RANTES (but not IL-4) were significantly higher in patients compared with controls. Highest concentrations were generally found during the febrile, hypotensive and oliguric disease phases, as well as in clinically severe and critical cases. CONCLUSION Serum concentrations of proinflammatory cytokines and chemokines increased in line with disease severity in HFRS patients.


title: The spread of pathogens through trade in poultry hatching eggs: overview and recent developments.

text: The international trade in poultry hatching eggs could potentially facilitate the global dissemination of poultry disease. Provided the guidelines of the World Organisation for Animal Health (OIE) on breeding flock hygiene are followed, of those avian diseases currently listed by the OlE, only highly pathogenic avian influenza (HPAI), Newcastle disease (ND), and avian mycoplasmosis (caused by Mycoplasma gallisepticum or M. synoviae) should be considered likely to be spread though trade in this commodity. Furthermore, the impact of HPAI and ND on egg production and hatchability will constrain the potential for these agents to be spread by poultry hatching eggs.


Is it just a marker for increased care?


title: [Epidemiological characteristics of severe acute respiratory syndrome in Tianjin and the assessment of effectiveness on measures of control].

text: OBJECTIVE To analyze the epidemiologic characteristics of severe acute respiratory syndrome (SARS) and to evaluate the effectiveness on its major control measures in Tianjn. METHODS Adopting two case reports 1 and 2, designed by the Tianjin Centers for Disease Control and Prevention to develop a unified case-tracing table including the map of the distribution of close contacts to SARS patients. With those methods, investigation on patients and their close contacts at hospital wards, families, communities and institutions of the patients were carried out. RESULTS From April 13 through May 8, 2003, there were 175 SARS cases including imported ones, were identified with an incidence rate of 1.9 cases per 100,000. Among them, 14 died with a fatality of 8.0%. The whole process of epidemic in Tianjin was less than one month with the following features: (1) 93.7% of the total SARS cases in Tianjin were directly or indirectly transmitted by a super-spreader. (2) 68.6% of the total SARS patients were concentrated in 3 hospitals A, B and C which was menifastated in 'clustering'. Through study on the rest of the SARS patients, results showed that 16.8% of them were transmitted through family close contact and 2.3% due to contact to colleagues. However, 12.6% of the patients were not able to show evidence that they had any contact to a diagnosed SARS patient. At the early stage of the epidemic, a number of medical practitioners were infected, taking up 38.2% of the total SARS cases. Among the total number of 1 975 medical workers who participated in the SARS medical cares and treatments, 3.4% of them got infected. During the outbreak, all index cases and chains of transmission seemed to be clear, with only 3 patients not able to be traced for the source of infection, taking up 2% of the total SARS patients in Tianjin. Among the 10 index cases, only the super spreader and another one index case transmitted the virus to their contacts but the rest of index cases did not cause any secondary infection. CONCLUSION Though SARS is clinically severe and can be spreaded quickly, the epidemic can be under control within a short period of time if chains of SARS transmission are broken down and effective measures as isolation and quarantine against patients as well as underscoring awareness among the publics in a scientific way, being carried out.


title: [Recent developments in SARS vaccine studies].

text: Severe acute respiratory syndrome (SARS) caused thousands of human infections worldwide and hundreds of deaths in just a few months. Evidence indicates that SARS coronavirus (SARS-CoV) has been circulating from animals to humans since before the 2002-2003 outbreak, suggesting that another pandemic may occur. This possibility has focused continuous action on SARS vaccine research. Inactivated vaccines, viral and bacterial vector vaccines, recombinant protein vaccines, subunit vaccines, DNA vaccines, and live-attenuated virus vaccines have been studied in different animal models. Although different animal models are used in vaccine studies, the most appropriate model for studying SARS is ferret since it develops the typical clinical signs, viral replication patterns and lung pathology compatible with that of SARS pathogenesis in humans. While there is much evidence that various vaccine strategies against SARS are safe and immunogenic, vaccinated animals still display significant disease upon challenge. Moreover, potential vaccine enhancement of SARS have also been shown in some studies. Data from the studies give an important information of the demand for further vaccine development research, especially focusing on mucosal immunization, T-cell immunity and combinations of heterologous vaccines in prime-boost regimens. In this review article developments on SARS vaccines have been discussed under the light of recent literature.


title: Canine parvovirus epidemiology in Bulgaria.

text: Canine parvovirus 2 (CPV-2) emerged in 1978 as one of the most pathogenic etiologic agents in dogs. Under the influence of evolution, the original CPV-2 was replaced, a few years later, by 2 variants, CPV-2a and CPV-2b. In 2000, a new variant, CPV-2c, was detected first in Italy and later in other countries. The current study was conducted to provide data about the CPV types circulating in Bulgaria. Forty-two fecal samples from dogs with clinical signs of parvovirosis, collected between June 2009 and February 2010, were tested for CPV using a rapid test based on detection of CPV antigens and a real-time polymerase chain reaction (PCR) for detection of viral DNA. Positive samples were characterized by means of minor groove binder probe PCR assays. Forty samples were positive, of which 30 were identified as CPV-2a, 9 as CPV-2b, and 1 as CPV-2c. The results from this molecular investigation of CPV show the prevalence of type 2a and occurrence of type 2c for the first time in Bulgaria.


title: Severe acute respiratory syndrome surveillance in Australia.

text: In March 2003, the World Health Organization (WHO) issued a global alert recommending active worldwide surveillance for severe acute respiratory syndrome (SARS). This paper describes the epidemiological features of cases reported by Australian states and territories to the Australian Government Department of Health and Ageing between 17 March and 31 July 2003. There were 138 people investigated for SARS: 111 as suspect and 27 as probable. Five probable cases were reported to WHO after review of other possible diagnoses and Australia-specific exclusion criteria had been applied. An additional probable case identified by laboratory testing overseas, but who was not under investigation when in Australia, was also reported to WHO. The method by which surveillance for SARS was rapidly established provided an opportunity to examine Australia's planning and preparedness for future respiratory disease epidemics such as influenza.


title: Bats: important reservoir hosts of emerging viruses.

text: Bats (order Chiroptera, suborders Megachiroptera ["flying foxes"] and Microchiroptera) are abundant, diverse, and geographically widespread. These mammals provide us with resources, but their importance is minimized and many of their populations and species are at risk, even threatened or endangered. Some of their characteristics (food choices, colonial or solitary nature, population structure, ability to fly, seasonal migration and daily movement patterns, torpor and hibernation, life span, roosting behaviors, ability to echolocate, virus susceptibility) make them exquisitely suitable hosts of viruses and other disease agents. Bats of certain species are well recognized as being capable of transmitting rabies virus, but recent observations of outbreaks and epidemics of newly recognized human and livestock diseases caused by viruses transmitted by various megachiropteran and microchiropteran bats have drawn attention anew to these remarkable mammals. This paper summarizes information regarding chiropteran characteristics and information regarding 66 viruses that have been isolated from bats. From these summaries, it is clear that we do not know enough about bat biology; we are doing too little in terms of bat conservation; and there remain a multitude of questions regarding the role of bats in disease emergence.


title: [Influence of physical activity during outbreak on psychological states in adults in the Covid-19 pandemic: a study protocol.]

text: This coronavirus pandemic has placed unprecedented restrictions on people's physical activity and routines. Prolonged home stays may lead to fear, panic, anxiety, and depression states, which in turn, can drive to a reduction of active lifestyles. Hence, determining the psychological response in the general population, and the influence level of home-based physical activity development could be relevant during this exceptional Covid-19 disease quarantine period. A multicenter, cross-sectional, and observational study design will be conducted in 12 Iberoamerican countries expecting to enroll 3,096 participants, through a snowball sampling technique. The study started on March 15th, 2020, and it is expected to be completed in August 2020 through online survey that will include demographic data, health status, psychological impact of the Covid-19 outbreak, mental health status, and level of physical activity. This study will be conducted following the principles established by the protocol, the Declaration of Helsinki, and the Ethical Guidelines for Clinical Research. Data from the study will be disseminated in manuscripts for submission to peer-reviewed journals as well as in abstracts for submission to relevant conferences. Trial registration number: NCT04352517, pre-results.


title: Influenza virus-like particles comprised of the HA, NA, and M1 proteins of H9N2 influenza virus induce protective immune responses in BALB/c mice.

text: Avian influenza viruses represent a growing threat for an influenza pandemic. To develop recombinant vaccine for avian influenza of the H9N2 subtype, we expressed in insect cells virus-like particles (VLPs) consisting of three structural proteins of influenza A/Hong Kong/1073/99 (H9N2) virus. Upon infection of Sf9 cells with recombinant baculoviruses, the hemagglutinin (HA), neuraminidase (NA), and matrix (M1) proteins were co-expressed in the infected cells, self-assembled, and released into the culture medium as VLPs of 80-120nm in diameter. VLPs exhibited functional characteristics of influenza virus including hemagglutination and neuraminidase activities. In BALB/c mice, VLPs elicited serum antibodies specific for influenza A/Hong Kong/1073/99 (H9N2) virus and inhibited replication of the influenza virus after challenge. Thus, VLPs represent a potential strategy for the development of human vaccines against avian influenza H9N2 viruses.


In vivo and in vitro models of demyelinating diseases.


title: Severe acute respiratory syndrome vaccine development: experiences of vaccination against avian infectious bronchitis coronavirus.

text: Vaccines against infectious bronchitis of chickens (Gallus gallus domesticus) have arguably been the most successful, and certainly the most widely used, of vaccines for diseases caused by coronaviruses, the others being against bovine, canine, feline and porcine coronaviruses. Infectious bronchitis virus (IBV), together with the genetically related coronaviruses of turkey (Meleagris gallopovo) and ring-necked pheasant (Phasianus colchicus), is a group 3 coronavirus, severe acute respiratory syndrome (SARS) coronavirus being tentatively in group 4, the other known mammalian coronaviruses being in groups 1 and 2. IBV replicates not only in respiratory tissues (including the nose, trachea, lungs and airsacs, causing respiratory disease), but also in the kidney (associated with minor or major nephritis), oviduct, and in many parts of the alimentary tract--the oesophagus, proventriculus, duodenum, jejunum, bursa of Fabricius, caecal tonsils (near the distal end of the tract), rectum and cloaca (the common opening for release of eggs and faeces), usually without clinical effects. The virus can persist, being re-excreted at the onset of egg laying (4 to 5 months of age), believed to be a consequence of the stress of coming into lay. Genetic lines of chickens differ in the extent to which IBV causes mortality in chicks, and in respect of clearance of the virus after the acute phase. Live attenuated (by passage in chicken embryonated eggs) IBV strains were introduced as vaccines in the 1950s, followed a couple of decades later by inactivated vaccines for boosting protection in egg-laying birds. Live vaccines are usually applied to meat-type chickens at 1 day of age. In experimental situations this can result in sterile immunity when challenged by virulent homologous virus. Although 100% of chickens may be protected (against clinical signs and loss of ciliary activity in trachea), sometimes 10% of vaccinated chicks do not respond with a protective immune response. Protection is short lived, the start of the decline being apparent 9 weeks after vaccination with vaccines based on highly attenuated strains. IBV exists as scores of serotypes (defined by the neutralization test), cross-protection often being poor. Consequently, chickens may be re-vaccinated, with the same or another serotype, two or three weeks later. Single applications of inactivated virus has generally led to protection of <50% of chickens. Two applications have led to 90 to 100% protection in some reports, but remaining below 50% in others. In practice in the field, inactivated vaccines are used in laying birds that have previously been primed with two or three live attenuated virus vaccinations. This increases protection of the laying birds against egg production losses and induces a sustained level of serum antibody, which is passed to progeny. The large spike glycoprotein (S) comprises a carboxy-terminal S2 subunit (approximately 625 amino acid residues), which anchors S in the virus envelope, and an amino-terminal S1 subunit (approximately 520 residues), believed to largely form the distal bulbous part of S. The S1 subunit (purified from IBV virus, expressed using baculovirus or expressed in birds from a fowlpoxvirus vector) induced virus neutralizing antibody. Although protective immune responses were induced, multiple inoculations were required and the percentage of protected chickens was too low (<50%) for commercial application. Remarkably, expression of S1 in birds using a non-pathogenic fowl adenovirus vector induced protection in 90% and 100% of chickens in two experiments. Differences of as little as 5% between the S1 sequences can result in poor cross-protection. Differences in S1 of 2 to 3% (10 to 15 amino acids) can change serotype, suggesting that a small number of epitopes are immunodominant with respect to neutralizing antibody. Initial studies of the role of the IBV nucleocapsid protein (N) in immunity suggested that immunization with bacterially expressed N, while not inducing protection directly, improved the induction of protection by a subsequent inoculation with inactivated IBV. In another study, two intramuscular immunizations of a plasmid expressing N induced protective immunity. The basis of immunity to IBV is not well understood. Serum antibody levels do not correlate with protection, although local antibody is believed to play a role. Adoptive transfer of IBV-infection-induced alphabeta T cells bearing CD8 antigen protected chicks from challenge infection. In conclusion, live attenuated IBV vaccines induce good, although short-lived, protection against homologous challenge, although a minority of individuals may respond poorly. Inactivated IBV vaccines are insufficiently efficacious when applied only once and in the absence of priming by live vaccine. Two applications of inactivated IBV are much more efficacious, although this is not a commercially viable proposition in the poultry industry. However, the cost and logistics of multiple application of a SARS inactivated vaccine would be more acceptable for the protection of human populations, especially if limited to targeted groups (e.g. health care workers and high-risk contacts). Application of a SARS vaccine is perhaps best limited to a minimal number of targeted individuals who can be monitored, as some vaccinated persons might, if infected by SARS coronavirus, become asymptomatic excretors of virus, thereby posing a risk to non-vaccinated people. Looking further into the future, the high efficacy of the fowl adenovirus vector expressing the IBV S1 subunit provides optimism for a live SARS vaccine, if that were deemed to be necessary, with the possibility of including the N protein gene.