title: Thoracoscopic Patch Repair of Congenital Diaphragmatic Hernia in a Neonate using Spiral Tacks: A Case Report.

text: We present a case of congenital diaphragmatic hernia that was successfully treated with spi-ral tacks using thoracoscopy. A newborn female was diagnosed with a diaphragmatic hernia at 20 weeks of gestation. The defect was surgically repaired by thoracoscopy and primary closure. On postoperative day 25, she developed respiratory distress. Chest x-ray showed a recurrence and was taken to the OR for surgical repair with spiral tacks.


title: Clinical characteristics of a Dutch DFNA9 family with a novel COCH mutation, G87W.

text: The present study aims to report audiological and vestibular characteristics of a Dutch DFNA9 family with a novel mutation, G87W, in the LCCL domain of COCH. From the family with the novel G87W COCH mutation audiometric data were collected and analyzed longitudinally. Results were compared with those obtained in previously identified P51S COCH mutation carriers (n = 74) and with those obtained in G88E mutation carriers. Special attention was also given to a comparison of age-related features, such as progressive hearing loss and vestibular impairment. A novel mutation (G87W) in COCH is indicative of hearing impairment and vestibular dysfunction in the present family. Pure-tone thresholds, phoneme recognition scores, and vestibular responses of the G87W mutation carriers were essentially similar to those previously established in the P51S and G88E mutation carriers. Deterioration of hearing and vestibular function in the G87W mutation carriers started at the age of 43 years. Remarkably, similar to G88E mutation carriers, the proportion of patients over 40 years of age who developed complete vestibular areflexia was significantly lower for the G87W mutation carriers than for the P51S mutation carriers. In conclusion, the phenotype associated with the novel COCH (G87W) mutation is largely similar to that associated with the P51S and G88E mutation carriers. However, subtle differences in terms of onset age and rate of progression seem to exist.


title: Localization of neutralizing epitopes and receptor-binding site in murine coronavirus spike protein.

text: To identify the localization of the epitopes recognized by monoclonal antibodies (MAbs) against the S1 subunit of the murine coronavirus JHMV spike protein, we have expressed the S1 proteins with different deletions from the C terminus of the S1. All of MAbs in groups A and B recognized the S1N(330) composed of 330 amino acids (aa) from the N terminus of the S1 and the larger S1 deletion mutants, but failed to react with the S1N(220) composed of 220 aa. MAbs in group C reacted only with the S1utt protein without any deletion. These results indicated that the S1N330 comprised the cluster of epitopes recognized by MAbs in groups A and B. These results together with the fact that all the MAbs in group B retained the high neutralizing activity suggested that the N terminus 330 aa are responsible for binding to the MHV-specific receptors. In pursuit of this possibility, we have expressed the receptor protein and examined the binding of each S1 deletion mutants to the receptor. It was demonstrated that the S1N(330) protein as well as other S1 deletion mutants larger than S1N(330) bound to the receptor. These results indicated that a domain composed of 330 aa at the N terminus of the S1 protein is responsible for binding to the MHV-specific receptor.


title: Predictability and preparedness in influenza control.

text: The threat of pandemic human influenza looms as we survey the ongoing avian influenza pandemic and wonder if and when it will jump species. What are the risks and how can we plan? The nub of the problem lies in the inherent variability of the virus, which makes prediction difficult. However, it is not impossible; mathematical models can help determine and quantify critical parameters and thresholds in the relationships of those parameters, even if the relationships are nonlinear and obscure to simple reasoning. Mathematical models can derive estimates for the levels of drug stockpiles needed to buy time, how and when to modify vaccines, whom to target with vaccines and drugs, and when to enforce quarantine measures. Regardless, the models used for pandemic planning must be tested, and for this we must continue to gather data, not just for exceptional scenarios but also for seasonal influenza.


Lack of trust hampers hunt for weapons.


title: Comparison of public health and preventive medicine physician specialty training in six countries: Identifying challenges and opportunities.

text: RATIONALE Public health and preventive medicine (PHPM) has been recognized internationally as a physician specialty, but national parallels and differences exist between training contexts. This paper reviews PHPM training and employment in Canada, France, Italy, Japan, the United Kingdom, and the USA. METHODS Information gathered from relevant accreditation bodies and literature searches was used to create descriptive profiles of national training demographics and structure and a narrative outlining trends and challenges facing the specialty. RESULTS Notable similarities and differences exist between national contexts. Key themes were differences in training strategies and practice scope, specialty stakeholders, certification structure, and funding. Recognition challenges faced the specialty across all six countries. Other challenges included unclear competencies and training strategies and a need for PHPM specialists to highlight their role in combating population health threats. Additional differences existed between comparator countries on the structure of training, funding sources for training programs, availability of training posts, and linkages with other physician specialties. CONCLUSION Highlighting these themes is a first step to fostering training collaborations between PHPM specialist physicians to augment transnational action on global public health challenges and also supports PHPM physician educators with innovative solutions from abroad that might address domestic specialty challenges.


title: [An epidemic outbreak of respiratory infection caused by Chlamydia pneumoniae in medical workers].

text: OBJECTIVE To investigate the clinical manifestations and the chest imaging characteristics of an epidemic outbreak of respiratory infection caused by Chlamydia pneumoniae (CP). METHODS A prospective study for CP infection in 15 patients from September 2003 was carried out. Sputum and throat swab specimen were obtained and CP DNA was detected by polymerase chain reaction (PCR). Serum samples were obtained and immunoglobulin G and M (IgG and IgM) of antibodies to CP. pneumoniae were studied by microimmunofluorescence test. Chest X-ray and computed tomography were retrospectively analyzed. RESULTS All patients presented fever, headache, sore throat, hoarseness, muscular ache, and dry cough. Acute cough was often associated with chest pain. The sputum blood was present in 3 patients (20%). Moist rales were heard in 4 patients. Chest imaging abnormalities were present in 67% (10 patients). The organism was demonstrated in 87% (13 patients) by PCR. The most common imaging abnormalities were unilateral and (or) bilateral multi-focal or solitary alveolar nodular opacities (9 patients). The patchy shadows were found in 2 patients, and pulmonary consolidation associated with the local pulmonary edema in 1 patient. Hilar or mediastinal lymphadenopathy and pleural effusion was not found. CONCLUSIONS The colony occurrences and similar clinical and chest imaging manifestations are characteristics of an outbreak of respiratory infection caused by CP in medical workers. An outbreak of respiratory infection caused by CP should be differentiated from severe acute respiratory syndrome (SARS).


title: Development and Validation of a Scoring System for Predicting Periprocedural Complications During Percutaneous Coronary Interventions of Chronic Total Occlusions: The Prospective Global Registry for the Study of Chronic Total Occlusion Intervention (PROGRESS CTO) Complications Score.

text: BACKGROUND High success rates are achievable for chronic total occlusion (CTO) percutaneous coronary intervention (PCI) using the hybrid approach, but periprocedural complications remain of concern. Although scores estimating success and efficiency in CTO PCI have been developed, there is currently no available score for estimation of the risk for periprocedural complications. We sought to develop a scoring tool for prediction of periprocedural complications during CTO PCI. METHODS AND RESULTS We analyzed data from 1569 CTO PCIs in the Prospective Global Registry for the Study of Chronic Total Occlusion Intervention (PROGRESS CTO) using a derivation and validation sampling ratio of 2:1. Variables independently associated with periprocedural complications in multivariable analysis in the derivation set were assigned points based on their respective odds ratios. Forty-four (2.8%) patients experienced complications. Three factors were independent predictors of complications and were included in the score: patient age >65 years, +3 points (odds ratio, OR=4.85, CI 1.82-16.77); lesion length ≥23 mm, +2 points (OR=3.22, CI 1.08-13.89); and use of the retrograde approach +1 point (OR=2.41, CI 1.04-6.05). The resulting score showed good calibration and discriminatory capacity in the derivation (Hosmer-Lemeshow χ2 6.271, P=0.281, receiver-operating characteristic [ROC] area=0.758) and validation (Hosmer-Lemeshow χ2 4.551, P=0.473, ROC area=0.793) sets. Score values of 0 to 2, 3 to 4, and ≥5 were defined as low, intermediate, and high risk of complications (derivation cohort 0.4%, 1.8%, 6.5%, P<0.001; validation cohort 0.0%, 2.5%, 6.8%, P<0.001). CONCLUSIONS The PROGRESS CTO complication score is a useful tool for prediction of periprocedural complications in CTO PCI. CLINICAL TRIAL REGISTRATION URL: http://www.clinicaltrials.gov. Unique identifier: NCT02061436.


ACE in COPD: a therapeutic target?


Trends in hospital admissions and mortality from asthma and chronic obstructive pulmonary disease in Australia.


title: The effects of land transport on animal welfare.

text: Animal welfare during and as a result of transport can be assessed by using a range of behavioural, physiological, pathological and carcass-quality indicators that are described in this paper. Measures of the extent of any disease, injury or mortality resulting from, or exacerbated by, transport are important because health is an important part of welfare. Many of the indicators are measures of stress as they involve long-term adverse effects on the individual. Factors affecting the welfare of animals before, during and after transport which are discussed are: definition of the responsibilities and competence, attitudes to animals and need for training of staff; planning of journeys and methods of payment of staff; laws and retailers' codes; genetics, especially selection for high productivity; rearing conditions and experience; the mixing of animals from different social groups; handling and loading procedures; driving methods; space allowance; increased susceptibility to disease and efforts to minimise the spread of disease.


title: Targeting glycosylation as a therapeutic approach.

text: Increased understanding of the role of protein- and lipid-linked carbohydrates in a wide range of biological processes has led to interest in drugs that target the enzymes involved in glycosylation. But given the importance of carbohydrates in fundamental cellular processes such as protein folding, therapeutic strategies that modulate, rather than ablate, the activity of enzymes involved in glycosylation are likely to be a necessity. Two such approaches that use imino sugars to affect glycosylation enzymes now show considerable promise in the treatment of viral infections, such as hepatitis B, and glucosphingolipid storage disorders, such as Gaucher disease.


title: Single hospital visit day case laparoscopic hernia repair without prior outpatient consultation is safe and acceptable to patients.

text: BACKGROUND For some common conditions, pre-operative clinic visits are often of little value to the patient or surgeon with transfer to the waiting list being predictable. In response to local patient feedback, we introduced a single hospital visit laparoscopic hernia surgery pathway with focus on informed consent, patient-reported outcomes and post-operative interaction with primary care services. METHODS A single hospital visit service for elective hernia repairs was created. Patients were not excluded on age, BMI or co-morbidity. Following referral, patients were telephoned by a surgeon. If considered appropriate, a symptom assessment tool, procedure information and consent form were sent. All patients were operated without attending clinic or pre-operative assessment. Surgeon-led telephone follow-up was made at either 2 or 7 days post-operatively and patient satisfaction assessed at 3 months. RESULTS A total of 517 patients were referred for single-stop surgery between 2012 and 2015. Median age was 58 (range 20-92), 91 % were male, and mean BMI was 25.6 (17.4-52.0). No patient refused the single-visit pathway. Single-stop patients had higher knowledge questionnaire scores (mean 16 vs. 10, p = 0.01) than patients who had attended clinic. Nine (1.7 %) were requested to attend clinic to confirm diagnosis, and three (0.8 %) were cancelled by their surgeon on the operative day. A total of 393 hernia repairs (331 TEP, 63 open) were performed under general anaesthetic. 92 % were discharged on day zero. Telephone follow-up day two rather than seven decreased attendance to primary care services (25 % vs. 57 %, p = 0.001). At 3 months, 95 % were satisfied and symptom scores were reduced (median 5-0, p < 0.0001). CONCLUSION Single-visit surgery appears to extend the patient benefits of laparoscopy by reducing hospital visits without compromising safety. Single hospital visit hernia surgery for unselected primary care referrals is possible and acceptable to patients.


Methods to identify increased risk of coronary disease in the general population: conclusion is oversimplification.


title: Patent ductus arteriosus in an adult amur leopard (Panthera pardus orientalis).

text: A clinically healthy 16-yr-old female leopard (Panthera pardus orientalis) was diagnosed with a patent ductus arteriosus on echocardiography and later confirmed on necropsy A murmur was heard on auscultation during a routine examination, and the congenital defect was an incidental finding. The animal had been asymptomatic its entire life. This deformity is rarely observed in nondomestic felids and may be asymptomatic, as has been described in domestic cats.


Severe acute respiratory syndrome (SARS): epidemiology, diagnosis and management.


title: Febrile respiratory illness in the intensive care unit setting: an infection control perspective.

text: PURPOSE OF REVIEW We have examined studies published since the severe acute respiratory syndrome outbreak that elucidate the mode of transmission of respiratory pathogens and the optimal means of interrupting their transmission, focusing on transmission in the intensive care unit. RECENT FINDINGS The severe acute respiratory syndrome may be transmitted by the droplet, contact and occasionally the airborne route. Transmission occurred most frequently when healthcare workers failed to use standard precautions such as hand washing or personal protective equipment designed to limit droplet and contact transmission. Aerosol-generating procedures increased transmission, but incomplete use of personal protective equipment during procedures was common. Recent publications have stressed the benefits of appropriate staffing levels, the use of single occupancy rooms, the immunization of healthcare workers against influenza, and the importance of healthcare workers remaining home when ill in reducing the morbidity and mortality of respiratory infection within the intensive care unit. SUMMARY Most respiratory pathogens can be transmitted by more than one route. Despite this, healthcare worker awareness of clinical syndromes associated with respiratory pathogens that require airborne precautions, combined with the use of standard precautions for all patients, and contact/droplet precautions for patients with undifferentiated febrile respiratory illness should be effective in interrupting the transmission of respiratory pathogens within the intensive care unit.


Infectious diseases. Viral DNA match spurs China's civet roundup.


title: Update: severe acute respiratory syndrome--United States, May 28, 2003.

text: CDC continues to work with state and local health departments, the World Health Organization (WHO), and other partners to investigate cases of severe acute respiratory syndrome (SARS). This report updates SARS cases reported worldwide and in the United States and reports a seventh laboratory-confirmed U.S. case.


title: Inter- and intra-variant genetic heterogeneity of human coronavirus OC43 strains in France.

text: Human coronavirus OC43 (HCoV-OC43) causes acute, self-limited respiratory infections. A close relationship between bovine coronaviruses (BCoVs) and HCoV-OC43 has recently been demonstrated. This study includes seven clinical, non-cell culture-adapted, contemporary HCoV-OC43 strains detected in France in 2003. By using RT-PCR and clonal sequencing of the S1 gene of HCoV-OC43, the inter-variant heterogeneity of the HCoV-OC43 circulating strains was studied and the intra-variant diversity was assessed by investigation of a quasispecies cloud. This paper brings to the forefront a high genetic diversity of circulating HCoV-OC43 variants. Genetically different groups are defined among the variants described in this study. One of these variants holds characteristics of an outlier and presents a deletion of 12 nt, also found in BCoV strains. Moreover, the presence of HCoV-OC43 as a quasispecies cloud in vivo during an acute respiratory-tract illness was discovered. It has also been revealed that quasispecies-cloud sizes are similar for the two viral populations tested.


title: Molecular cloning and expression analysis of bat toll-like receptors 3, 7 and 9.

text: In this study, cDNA of Toll-like receptors (TLR) 3, 7 and 9 were synthesized and completely sequenced. The coding regions of cDNA for bat TLR3, TLR7 and TLR9 were 2,718, 3,150 and 3,090 bp in length, respectively. The open reading frames encoded 905, 1,049 and 1,029 amino acids for TLR3, TLR7 and TLR9, respectively. The nucleotide sequences, predicted amino acid sequences and predicted domain structures of the three bat TLRs had high homology with those of other mammals. In addition, the expression profiles of each TLR in main organs were analyzed. Expression of TLR3 was highest in the liver, whereas the expressions of TLR7 and TLR9 were highest in the spleen.


title: Novel coronavirus disease (COVID-19) outbreak: Now is the time to refresh pandemic plans.

text: This article outlines practical steps that businesses can take now to prepare for a pandemic. Given the current growing spread of coronavirus disease 2019 (COVID-19) around the world, it is imperative that businesses review their pandemic plans and be prepared in case this epidemic expands and affects more people and communities. Preparing for a potential infectious disease pandemic from influenza or a novel corona virus is an essential component of a business continuity plan, especially for businesses that provide critical healthcare and infrastructure services. Although many businesses and organisations have a pandemic plan or address pandemic preparedness in their business continuity plans, few have recently tested and updated their plans. Pandemics can not only interrupt an organisation's operations and compromise long-term viability of an enterprise, but also disrupt the provision of critical functions. Businesses that regularly test and update their pandemic plan can significantly reduce harmful impacts to the business, play a key role in protecting employees' and customers' health and safety, and limit the negative impact of a pandemic on the community and economy.


title: [Options for controlling new Corona virus infection - 2019-nCoV].

text: According to the Situation Report 65 of the World Health Organization of March 25, 2020, the COVID-19 incidence rate indicates 413 467 confirmed cases and 18 433 deaths. Genetic diversification of the Corona virus has resulted in strains that cause severe respiratory tract infections in humans via drip and animal mediation. S-proteins covering its surface, which bind to the cell receptor - angiotensin converting enzyme 2 (ACE-2) and transmembrane serine protease (TMPRSS2) are important in shaping virus activity. The course of infection varies from mild to severe. The ability to control infection is limited because there are no drugs that fully inhibit 2019-nCoV. Interferon-alpha (5 million U twice daily by inhalation), lopinavir/ritonavir (400/100 mg twice daily orally), as well as chloroquine (500 mg twice daily orally for 10 days) and azithromycin (500 mg twice per day) cause a milder course of the disease and reduce the duration of treatment. The administration of glucocorticosteroids and research drugs (tocilizumab) is acceptable for massive infiltrative lesions in the pulmonary parenchyma causing severe lung injury (ALI) and acute respiratory distress syndrome (ARDS). In the system operation it is necessary to create the socalled a safety matrix that would take into account the existing threat on the one hand and all available services and resources on the other. Precise analysis and separation of individual tasks can enable the creation of a real crisis management plan.


title: Human adenoidal organ culture: a model to study nontypable Haemophilus influenzae (NTHI) and other bacterial interactions with nasopharyngeal mucosa--implications in otitis media.

text: Nontypable Haemophilus influenzae (NTHI) has become the predominant cause of both acute suppurative otitis media and chronic otitis media with effusion. It has now been well-demonstrated that both outer membrane proteins and restriction fragment analysis of the bacterial genomes of concomitant nasopharyngeal and middle ear effusion isolates of NTHI are identical. It is therefore of critical importance to understand the mechanisms whereby bacteria that are present in normal healthy children in small numbers become the predominant organism in the nasopharynx in otitis media. The studies presented here suggest that nontypable Haemophilus influenzae can effectively decrease ciliary function as measured by stroboscopic illumination of ciliary beat frequency on human adenoidal organ culture. This organism also produces significant histopathologic and ultrastructural damage to the epithelial cells and cilia of adenoid organ culture, demonstrated by both light microscopy and scanning electron microscopy. The data suggest the following hypothesis: nontypable Haemophilus influenzae can destroy mucociliary function and allow increased bacterial replication in the mucus overlying the nasopharyngeal mucosa. The mucociliary system of the eustachian tube may also be involved in a similar manner, thus allowing bacteria to enter the middle ear space via the eustachian tube.


title: Mortality among patients with tuberculosis and associations with HIV status --- United States, 1993-2008.

text: Worldwide, tuberculosis (TB) incidence increased from 125 cases per 100,000 population in 1990 to 142 cases per 100,000 population in 2004, primarily because of the human immunodeficiency virus (HIV) epidemic. Persons with HIV are at increased risk for TB disease, and those with TB have a high risk for death. This is documented most clearly in resource-limited settings, where limited access to antiretroviral therapy (ART) and other health-care services contribute to the elevated mortality. The impact of HIV on patients with TB is less clear in resource-rich nations such as the United States. To understand the impact of HIV on the risk for death during TB treatment in the United States, data were analyzed for all culture-positive patients with TB from 1993 to 2008, and the proportion that died was determined and stratified by HIV test result. Mortality data were restricted to patients reported before 2007. The proportion of all patients with TB who died during TB treatment decreased from 2,445 of 13,629 (18%) in 1993 to 682 of 7,578 (9%) in 2006. Among patients with TB and HIV, 950 of 2,337 (41%) died during treatment in 1993; this proportion declined to 131 of 663 (20%) in 2006. The proportion of patients with TB and HIV who received their TB diagnosis postmortem dropped from 191 of 2,927 (7%) in 1993 to 32 of 768 (4%) in 2006; 624 of 10,468 (6%) persons with TB and unknown HIV status received their TB diagnosis postmortem in 1993, and this proportion did not decline. Further reductions in mortality can be achieved by enhanced TB/HIV program collaboration and service integration.


title: What are the roles and responsibilities of the media in disseminating health information?

text: BACKGROUND TO THE DEBATE In December 2004 three news stories in the popular press suggested that the side effects of single-dose nevirapine, which has been proven to prevent mother-to-child transmission of HIV, had been covered up. Many HIV experts believed that the stories were unwarranted and that they would undermine use of the drug, leading to a rise in neonatal HIV infection. The controversy surrounding these stories prompted the PLoS Medicine editors to ask health journalists, and others with an interest in media reporting of health, to share their views on the roles and responsibilities of the media in disseminating health information.


title: [Experimental coronavirus encephalomyelitis in mice].

text: A combined comparative virological, morphological, and immunological study of experimental coronavirus encephalomyelitis was carried out in mice in order to elucidate the pathogenetic mechanisms involved in the formation of the foci of lesions in acute and chronic forms of the disease. Intracerebral inoculation of C3H mice with the neurotropic JHM strain of murine hepatitis virus induces a disease with demyelinization foci in the CNS running acute, subacute, or chronic course. This model underlies a concept that demyelinating diseases are caused by viruses producing immunopathologic responses realized via certain histocompatibility loci. The long-term persistence of viral antigen in the CNS and liver may be explained by virus replication in hepatocytes and oligodendrocytes at low levels, and exacerbations of the disease are prevented by the immune system.


Severe acute respiratory syndrome: private hospital in Singapore took effective control measures.


title: TNF-alpha-induced cell death in feline immunodeficiency virus-infected cells is mediated by the caspase cascade.

text: TNF-alpha induced apoptosis in a feline fibroblastic cell line (CRFK) infected with FIV but not in its uninfected control. In this study, to understand the molecular basis of the different susceptibilities to TNF-alpha between FIV-infected and uninfected cells, we examined the expression of TNF receptors and the activation of the caspase and NF-kappaB pathways. Expression levels of TNFR I and TNFR II mRNAs were similar between uninfected and FIV-infected CRFK cells. To understand the role of caspases in TNF-alpha-induced apoptosis, we examined the effect of three different classes of caspase inhibitors, Z-VAD-FMK, Ac-YVAD-CMK, and Z-DEVD-FMK, on the TNF-alpha-induced apoptosis in FIV-infected cells. Pretreatment with each of these caspase inhibitors protected FIV-infected CRFK cells from TNF-alpha-induced cell death. Moreover, one of the caspase substrates, poly(ADP-ribose) polymerase, was shown to be cleaved after TNF-alpha treatment in FIV-infected CRFK cells but not in uninfected CRFK cells. Electrophoretic mobility shift assay using an NF-kappaB motif oligonucleotide and promoter assay using an NF-kappaB luciferase reporter construct indicated that TNF-alpha treatment had induced activation of NF-kappaB in both FIV-infected and uninfected CRFK cells. The present study indicates that TNF-alpha-induced apoptosis in FIV-infected CRFK cells is mediated by the activation of the caspase cascade, but not by either upregulation of TNF receptor or inhibition of NF-kappaB.


title: Detection of turkey coronavirus in commercial turkey poults in Brazil.

text: Poult enteritis complex has been incriminated as a major cause of loss among turkey poults in other countries. We have observed this in Brazil, associated with diarrhoea, loss of weight gain and, commonly, high mortality. In this study, we have used the reverse transcriptase polymerase chain reaction (RT-PCR) to detect turkey coronavirus (TCoV) in sick poults 30 to 120 days of age from a particular producer region in Brazil. The RT-PCR was applied to extracts of intestine tissue suspensions, and the respective intestinal contents, bursa of Fabrícius, faecal droppings and cloacal swabs. Primers were used to amplify the conserved 3' untranslated region of the genome, and the nucleocapsid protein gene of TCoV. Histopathological and direct immunohistochemical examinations were performed to detect TCoV antigen in infected intestine and bursa slides. All the results from stained tissues revealed lesions as described previously for TCoV infection. The direct immunohistochemical positive signal was present in all intestine slides. However, all bursa of Fabrícius tissues analysed were negative. RT-PCR findings were positive for TCoV in all faecal droppings samples, and in 27% of cloacal swabs. Finally, the best field material for TCoV diagnosis was faecal droppings and/or intestine suspensions.


title: Serological study of coronavirus antibodies in Hamburg.

text: A serological study of coronaviruses with the OC 43 agent (HI-test) was undertaken in 1000 sera from the population of Hamburg. 32,7% of the sera showed HI=titers between 1 : 8 and 1 : 128. The geometric mean titer (GMT) for the whole group was 1 : 16. The frequency of seropositive reactions was highest in 244 individuals 15 to 24 years old (52,8% positive reactions, GMT 1 : 19) whilst in the older age groups (25-59 years and over 60 years old) the precentage of positive reactors was markedly lower (14,4% and 16,8%, respectively, GMT 1 : 12). Among 131 twelve to 17 years old youngsters, a seroconversion was demonstrated in paired sera in 6,8% with a 4- to 8-fold rise in HI-titer within 8 weeks.


title: Virus receptors: implications for pathogenesis and the design of antiviral agents.

text: A virus initiates infection by attaching to its specific receptor on the surface of a susceptible host cell. This prepares the way for the virus to enter the cell. Consequently, the expression of the receptor on specific cells and tissues of the host is a major determinant of the route of entry of the virus into the host and of the patterns of virus spread and pathogenesis in the host. This review emphasizes the virus-receptor interactions of human immunodeficiency virus, the rhinoviruses, the herpesviruses, and the coronaviruses. These interactions are often found to be complex and dynamic, involving multiple sites or factors on both the virus and the host cell. Also, the receptor may play an important role in virus entry per se in addition to its role in virus binding. In the cases of human immunodeficiency virus and the rhinoviruses, ingenious approaches to therapeutic strategies based on inhibiting virus attachment and entry are under development and in clinical trials.


title: First detected human bocavirus in a Malaysian child with pneumonia and pre-existing asthma: a case report.

text: Human bocavirus (HBoV) is a newly discovered parvovirus associated with respiratory disease in children. There are many reports worldwide on the endemicity of this virus. Since it is relatively new, detection in clinical laboratories is not routinely performed. We describe the first detection of HBoV in Malaysia in a 13-month-old boy with pneumonia and underlying asthma. The infective agent was confirmed by molecular methods.


title: Characterization of a human coronavirus (strain 229E) 3C-like proteinase activity.

text: The RNA polymerase gene of human coronavirus (HCV) 229E encodes a large polyprotein that contains domains with motifs characteristic of both papain-like cysteine proteinases and proteinases with homology to the 3C proteinase of picornaviruses. In this study, we have, first, expressed the putative HCV 229E 3C-like proteinase domain as part of a beta-galactosidase fusion protein in Escherichia coli and have shown that the expressed protein has proteolytic activity. The substitution of one amino acid within the predicted proteinase domain (His-3006-->Asp-3006) abolishes, or at least significantly reduces, this activity. Amino-terminal sequence analysis of a purified, 34-kDa cleavage product shows that the bacterial fusion protein is cleaved at the dipeptide Gln-2965-Ala-2966, which is the predicted amino-terminal end of the putative 3C-like proteinase domain. Second, we have confirmed the proteolytic activity of a bacterially expressed polypeptide with the amino acid sequence of the predicted HCV 229E 3C-like proteinase by trans cleavage of an in vitro translated polypeptide encoded within open reading frame 1b of the RNA polymerase gene. Finally, using fusion protein-specific antiserum, we have identified a 34-kDa, 3C-like proteinase polypeptide in HCV 229E-infected MRC-5 cells. This polypeptide can be detected as early as 3 to 5 h postinfection but is present in the infected cell in very low amounts. These data contribute to the characterization of the 3C-like proteinase activity of HCV 229E.


title: Cerebral Infarction by Paradoxical Gas Embolism During Laparoscopic Liver Resection with Injury of the Hepatic Vessels in a Patient without a Right-to-Left Systemic Shunt.

text: BACKGROUND Carbon dioxide (CO2) is believed to be the safest gas for laparoscopic surgery, which is a standard procedure. We experienced severe cerebral infarction caused by paradoxical CO2 embolism during laparoscopic liver resection with injury of the hepatic vessels despite the absence of a right-to-left systemic shunt. CASE REPORT A 60-year-old man was diagnosed with hepatocellular carcinoma in the right hepatic lobe secondary to alcoholic liver disease. We planned the laparoscopy-assisted liver resection. During the surgery, the root of the right hepatic vein was injured. A 1.5-cm hole was accidentally made in the right hepatic vein, while mobilizing the right hepatic lobe laparoscopically. End-tidal CO2 dropped from 39 to 15.5 mmHg, and systemic blood pressure dropped from 121 to 45 mmHg, returning to normal with the administration of inotropes. The transesophageal echocardiography revealed numerous bubbles in the left atrium and ventricle. The Bispectral Index monitoring system showed low brain activity, suggesting cerebral infarction due to paradoxical gas embolism. The hepatectomy was completed by conversion to open laparotomy. The patient went into a coma and suffered quadriplegia after surgery, despite the cooling of his head and the administration of Thiamylal. Brain MRI revealed cerebral infarction in the broad area of the cerebral cortex right side predominantly, with poor blood flow confirmed by the brain perfusion single-photon emission CT. Rehabilitation was gradually achieved with Botox injections. CONCLUSIONS Cerebral infarction by paradoxical gas embolism is a rare complication in laparoscopic surgery, but it is important to be aware of the risk and to be prepared to treat it.


title: The use of highly cross-linked polyethylene in total knee arthroplasty.

text: Polyethylene wear, with resultant particle-induced osteolysis, is a cause of late failure of total knee arthroplasty. The causes of both wear and osteolysis are multifactorial; still, improvements in the polyethylene liner have been investigated. Available highly cross-linked polyethylene tibial liners and patellar prostheses differ greatly in the amount and method of irradiation, thermal treatments, and sterilization techniques they undergo. Several varieties of highly cross-linked polyethylene reduce the gravimetric and volumetric wear of tibial liners in knee simulator studies. However, reduced fracture toughness and the generation of smaller and possibly more reactive particles also have been reported with some varieties of polyethylene. Clinical studies of the use of highly cross-linked polyethylene in total knee arthroplasty are limited. Two nonrandomized trials of highly cross-linked polyethylene in total knee arthroplasty have reported a nonsignificant decrease in radiolucent lines at 2 and 5 years, respectively. The risks of using highly cross-linked polyethylene include fracture of the liner or of a posterior-stabilized tibial post, liner dislodgement or locking mechanism disruption, and possibly more osteolysis. Highly cross-linked polyethylene tibial liners may be considered for younger, more active patients. However, until additional clinical results are available, a cautious approach is warranted to the widespread use of highly cross-linked polyethylene in total knee arthroplasty.


title: A prospective randomized trial comparing subatmospheric wound therapy with a sealed gauze dressing and the standard vacuum-assisted closure device: a supplementary subgroup analysis of infected wounds.

text: INTRODUCTION Subatmospheric pressure wound therapy (SAWT) is commonly used to manage infected wounds. However, this practice remains controversial because the safety and efficacy of the technique has not been carefully documented. METHODS The authors assessed the safety and efficacy of a sealed gauze dressing with wall suction applied (GSUC) compared to vacuum assisted-closure (VAC), both soaked with topical antimicrobials. Subjects included 31 hospitalized patients with acutely infected wounds compared with 56 patients with noninfected wounds. RESULTS There were significant reductions in wound surface area and volume in both infected and noninfected groups; there was no significant difference in the rate of change observed in the GSUC vs the VAC arms of the study. In the infected group, the reduction in wound surface area was 4.4% per day for GSUC and 4.8% per day for VAC. Wound volume was 7.8% per day for GSUC, and 9.7% per day for VAC (P < 0.001 for all). Evidence of wound infection in all patients, regardless of treatment group, resolved by 96 hours of onset of treatment, and there were no complications specifically related to the use of a sealed dressing over infected wounds. CONCLUSION Gauze dressing with wall suction and VAC therapy can be used in selected acute, infected wounds and both methods of treatment appear to be similarly effective for reducing wound surface area and volume. .


title: VP35 knockdown inhibits Ebola virus amplification and protects against lethal infection in mice.

text: Phosphorodiamidate morpholino oligomers (PMO) are a class of uncharged single-stranded DNA analogs modified such that each subunit includes a phosphorodiamidate linkage and morpholine ring. PMO antisense agents have been reported to effectively interfere with the replication of several positive-strand RNA viruses in cell culture. The filoviruses, Marburg virus and Ebola virus (EBOV), are negative-strand RNA viruses that cause up to 90% lethality in human outbreaks. There is currently no commercially available vaccine or efficacious therapeutic for any filovirus. In this study, PMO conjugated to arginine-rich cell-penetrating peptide (P-PMO) and nonconjugated PMO were assayed for the ability to inhibit EBOV infection in cell culture and in a mouse model of lethal EBOV infection. A 22-mer P-PMO designed to base pair with the translation start site region of EBOV VP35 positive-sense RNA generated sequence-specific and time- and dose-dependent inhibition of EBOV amplification in cell culture. The same oligomer provided complete protection to mice when administered before or after an otherwise lethal infection of EBOV. A corresponding nonconjugated PMO, as well as nonconjugated truncated versions of 16 and 19 base residues, provided length-dependent protection to mice when administered prophylactically. Together, these data suggest that antisense PMO and P-PMO have the potential to control EBOV infection and are promising therapeutic candidates.


title: The child's experience of single room isolation: a literature review.

text: Studies have shown that people who require single room isolation while in hospital often feel lonely, sad, worried, bored and in need of information. A literature review identified only 16 papers reporting on the child's experience of isolation. Findings indicate that children feel lonely, are scared of the personal protective equipment and feel bored. Patients' parents feel guilty, worried and under increased pressure to visit their children. It is also suggested that isolation may affect child development.


title: Protecting the waters of Wisconsin from microbiological threats.

text: Wisconsin is blessed with an abundance of high quality water that has played a significant role in the economic and social development of the state. The quality of life for Wisconsinites is often at least partially defined by pristine inland lakes, a portion of the Great Lakes shoreline, or great-tasting well water. While no one likes to associate water with disease, especially waters like those we enjoy in Wisconsin, the historical reality is that there is a strong connection. Until the 1920s, intestinal diseases such as typhoid fever and cholera often spread through water and were a leading cause of death in the United States. While easy access to modern medical care combined with improved water treatment technologies have greatly decreased waterborne disease, the microbiology of Wisconsin's waters still deserves continued vigilance and attention. The World Health Organization currently estimates that, on a global basis, 4 billion annual cases of waterborne diarrhea result in more than 2 million deaths per year--the equivalent of 20 jumbo jet crashes per day. Most of these deaths are in children under 5 years of age. The daily global death toll from waterborne disease is at least 1000 times greater than from the Severe Acute Respiratory Syndrome epidemic that recently made headline news. Waterborne disease is not just a problem in underdeveloped countries. Scientists who study disease transmission believe that 10%-30% of the vomiting and diarrhea illness in North America, including Wisconsin, may be acquired from water.


Morgagni Hernia: Is It Just a Congenital Hernia or a Cause of Emergency Surgery?


Ethics and SARS: lessons from Toronto.


title: [Construction of genomic full-length cDNA of SARS coronavirus BJ01 strain and identification of its biological characteristics].

text: Severe acute respiratory syndrome (SARS) is an important emerging infectious disease which caused by SARS coronavirus (SARS-CoV), and the study of its pathogenesis is needed for the treatment and prevention of this disease. To study the pathogenesis of SARS-CoV using reverse genetics technology, by in vitro ligation using 7 contiguous cDNAs that span the entire genome of the SARS-CoV BJ01 strain, a genomic full-length cDNA was assembled, then using T7 RiboMAX Large Scale RNA Production Systems with the genomic full-length cDNA as template, the RNA transcript was attained. The typical SARS-CoV-resulted cell pathogenic effects were observed when RNA transcript was electroporated into Vero E6 cells. The results of RT-PCR and sequencing of the rescued virus showed that it originated from transcript which derived from the full-length cDNA construct. Rescued virus-infected cells were detected by indirect fluorescent antibody staining demonstrated that it can specifically reaction with SARS-CoV. By CPE method and plaque assay, the titers of the rescued virus and wild-type virus were assessed, which demonstrated there are no significant difference between the viruses and they have similar biological characteristics. Construction of the genomic full-length cDNA of SARS-CoV BJ01 stain successfully and study of the biological characteristics of the rescued virus will provide a useful tool serving for the discovery of molecular pathogenesis and development of candidate vaccines against SARS-CoV.


title: Three men, a paint brush and a coronavirus.

text: Coronaviruses cause respiratory tract infection and a coryzal syndrome. Although described as a cause of gastroenteritis in HIV patients, with the exception of the severe acute respiratory syndrome (SARS), there is little in the literature about respiratory infection in HIV patients. We describe two patients with HIV, exacerbations of chronic obstructive pulmonary disease and proven coronavirus infection. A third patient presented with an upper respiratory tract infection but coronavirus was not isolated. All three men had spent a day decorating the first patient's flat four days prior to presentation. This is the first description of respiratory tract infection with coronavirus in HIV patients. Both patients with coronavirus required prolonged admission to hospital and extensive investigations because they were HIV infected. Coronavirus is often associated with less severe upper respiratory tract infection but can cause more severe disease and should be considered in patients with HIV and respiratory tract infection.


title: Influenza in the Pacific.

text: Influenza A and B viruses cause significant human disease worldwide through regular outbreaks and epidemics of seasonal influenza, and occasional pandemics when a novel influenza A virus emerges. Whereas Australia and New Zealand have well-established systems for community and laboratory-based surveillance of influenza, most other countries of the Pacific are only beginning to develop such systems with the support of various global and regional agencies and networks. Here we describe the role of the World Health Organization Global Influenza Surveillance Network and other organizations in laboratory-based influenza surveillance in the region and review some of the available data on seasonal and pandemic influenza in the developed and developing countries of the Pacific. The particular features of the Pacific Island countries and territories as small dispersed island communities, together with the greater susceptibility of indigenous people to the severe effects of influenza, highlight the importance of developing local laboratory-based surveillance systems. Such systems will improve the understanding, detection and control of seasonal influenza while also providing early warning of the emergence of potential pandemic viruses.


title: [An epidemiologic investigation on infection with severe acute respiratory syndrome coronavirus in wild animals traders in Guangzhou].

text: OBJECTIVE To investigate status of infection with severe acute respiratory syndrome coronovirus (SARS-CoV) in traders of wild animals wholesale markets in Guangzhou. METHODS Serum antibody against SARS-CoV IgG was determined cross-sectionally and symptoms of respiratory infection were investigated retrospectively for part of traders of three wholesale markets for wild animals in Guangzhou. RESULTS Overall rate of infection with SARS-CoV in 635 traders was 16.69%, varying in three different markets. Infection rate in market A mainly engaging in wild animals ranked the highest of 25.61%, significantly higher than that in markets B and C engaging in domestic fowls and snakes. Infection rate in traders only engaging in civet cats was 58.54%, significantly higher than that in traders engaging in snakes only (9.46%). In market A, infection rate varied in different persons, 59.34%, 20.59%, 16.00%, 15.22%, 10.40% and 9.68% in traders engaging in wild animals, managers, children of the traders, traders engaging in domestic fowls, traders engaging in snakes, and traders engaging in frozen food, respectively, in a decreasing pattern as their contact opportunities. During the period of SARS epidemic, detection rate of SARS-CoV antibody in people with symptoms of acute respiratory infection was higher (30.70%) than that in those without such symptoms (20.08%). Prevalence of symptoms of acute upper respiratory infection in people with positive antibody against SARS-CoV was higher (49.28%) than that in those with negative antibody (30.35%). CONCLUSIONS Infection with SARS-CoV in traders of animal markets possibly related to their direct exposure to wild animals, particularly to civet cats. During the period of SARS epidemic, some of the traders did infect with SARS-CoV, but they were neglected due to clinically inapparent manifestations.


title: [Evaluation of the Benefits of Administering Hand Therapy to Patients with Chemotherapy-Induced Peripheral Neuropathy].

text: Taxanes, which are used to treat breast cancer, damage the microtubules of normal nerve cells, causing numbness of the fingers related to chemotherapy-induced peripheral neuropathy(CIPN); therefore, effective methods for reducing numbness are needed. In 2017, it was reported that physical stimuli related to massage improved finger blood flow volumes, contributing to the regeneration of damaged nerves. We developed a method of hand therapy for breast cancer patients complaining of numbness related to anticancer drug administration, and examined its effects on numbness. Hand therapy was performed by a single therapist who received lectures at the Sophia Phytotherapy College, which is accredited by the Japan Handcare Association. The fingertips to wrist, ankle, metacarpal bones, palm, and elbow were massaged using the bilateral arms/fingers for 15minutes. We investigated the influences of daily living status(Support Team Assessment Sched- ule-Japan: STAS-J), age, body mass index(BMI), severity/site of numbness, type of numbness, type of drug, duration of breast cancer, duration of numbness, and presence or absence of lymph node dissection, and evaluated the severity of numbness using a 10-cm Visual Analog Scale(VAS). The study included 51 breast cancer patients complaining of numbness of the fingers, with a mean age of 59 years. In patients with relatively mild numbness(STAS-J 1), the VAS scores before and after hand therapy were 4.7±1.8 and 1.9±1.3, respectively, showing a marked decrease. In STAS-J 2 patients, the values were 4.9 ±1.4 and 2.1±1.3, respectively, also showing a marked decrease. Thus, this hand therapy reduced numbness in mild- and moderate-status patients. Statistical comparisons were performed between the STAS-J 1/2(mild/moderate numbness)and STAS-J 3/4(severe numbness)groups. Although the severity of numbness was not correlated with age, BMI, type of drug, lymph node dissection, or duration of breast cancer, the proportion of patients with a B1-year history of numbness was significantly larger in the STAS-J 1/2 group. The most frequent site of numbness was from the proximal interphalangeal joints to the fingertips. Concerning the severity of numbness, many patients complained of severe numbness, as represented by that after sitting straight. These results suggest that this hand therapy is effective for reducing numbness in patients receiving taxanes and complaining of mild to moderate numbness.


title: Pharmacologic treatment of SARS: current knowledge and recommendations.

text: The severe acute respiratory syndrome (SARS) pandemic caught the world by surprise in 2003 and spread rapidly within a relatively short period of time. Hence, randomised placebo-controlled clinical trials on the treatment of SARS were not possible. Our understanding was obtained from observational, cohort studies, case series and reports. Nevertheless, such information is useful in providing clinical management guidelines and directing future research in case SARS recurs. Early in the pandemic, a combination of ribavirin and corticosteroids was adopted as the standard treatment in Hong Kong, Canada and elsewhere because of the apparent good results of the first few patients. Subsequent reports showed that ribavirin was associated with a high rate of toxicity and lacked in vitro antiviral effect on SARS-coronavirus (SAR-CoV). The timing and dosage regimens of steroid in the treatment of SARS are controversial. Pulse methylprednisolone 250 to 500 mg/day for 3 to 6 days has been reported to have some efficacy in a subset of patients with "critical SARS", i.e., critically ill SARS patients with deteriorating radiographic consolidation, increasing oxygen requirement with PaO2 <10 kPa or SpO2 <90% on air, and respiratory distress (rate of 30/min). Prolonged therapy with high-dose steroids, in the absence of an effective antimicrobial agent, could predispose patients to complications such as disseminated fungal infection, and avascular necrosis. Kaletra (400 mg ritonavir and 100 mg lopinavir), a protease inhibitor used in the treatment of human immunodeficiency virus infection, may be considered for early treatment of SARS patients, preferably in a randomised double-blind placebo-controlled clinical trial setting. Interferon (IFN) is not recommended as standard therapy in SARS. However, there are enough data on in vitro activity of IFN preparations and a few clinical studies for these products to support a controlled trial if SARS recurs. Many other experimental treatments have been tried in an uncontrolled manner, and they should not be recommended as standard therapy.


Bilateral Infiltrates: Not Only COVID-19.


title: Lethal infection of K18-hACE2 mice infected with severe acute respiratory syndrome coronavirus.

text: The severe acute respiratory syndrome (SARS), caused by a novel coronavirus (SARS-CoV), resulted in substantial morbidity, mortality, and economic losses during the 2003 epidemic. While SARS-CoV infection has not recurred to a significant extent since 2003, it still remains a potential threat. Understanding of SARS and development of therapeutic approaches have been hampered by the absence of an animal model that mimics the human disease and is reproducible. Here we show that transgenic mice that express the SARS-CoV receptor (human angiotensin-converting enzyme 2 [hACE2]) in airway and other epithelia develop a rapidly lethal infection after intranasal inoculation with a human strain of the virus. Infection begins in airway epithelia, with subsequent alveolar involvement and extrapulmonary virus spread to the brain. Infection results in macrophage and lymphocyte infiltration in the lungs and upregulation of proinflammatory cytokines and chemokines in both the lung and the brain. This model of lethal infection with SARS-CoV should be useful for studies of pathogenesis and for the development of antiviral therapies.


title: Laparoendoscopic Single-Site (LESS) versus Robotic "Redo" Hiatal Hernia Repair with Fundoplication: Which Approach Is Better?

text: Only a small percentage of patients fail laparoscopic fundoplications undertaken for gastroesophageal reflux disease. But because many laparoscopic fundoplications have been undertaken, surgeons frequently encounter patients in need of "redo" operations. This study was undertaken to evaluate the robotic approach versus laparoendoscopic single-site (LESS) approach for redo fundoplications. With an Institutional Review Board approval, 64 patients undergoing LESS (n = 32) or robotic (n = 32) redo antireflux operations were prospectively followed up. Data are presented as median (mean + SD). For LESS versus robotic redo operations, the operative duration was 145 (143 ± 33.5) versus 196 (208 ± 76.7) minutes (P < 0.01), estimated blood loss was 50 (80 ± 92.1) versus 20 (43 ± 57.1) mL (P = 0.07), and length of stay was 1 (3 ± 5.4) versus 1 (2 ± 1.9) day (P = 0.57); 1 LESS operation was converted to "open." Operative duration was longer for men (P = 0.01). Postoperative complications were not more frequent after Nissen (n = 36) or Toupet (n = 28) fundoplication, regardless of the approach. When matched by BMI, operative duration was prolonged by a large Type I to IV hiatal hernia (P = 0.01). Symptoms improved dramatically and were similar with both approaches, and patient satisfaction was high. Robotic redo antireflux operations take longer than LESS operations. LESS and robotic redo antireflux operations are both safe and offer significant and similar amelioration of symptoms after failed fundoplications.


title: Epidemiology, surveillance and control of Nipah virus infections in Malaysia.

text: The outbreak of Nipah virus, affecting pigs and pig-farm workers, was first noted in September 1998 in the north-western part of peninsular Malaysia. By March 1999, the outbreak had spread to other pig-farming areas of the country, inclusive of the neighbouring country, Singapore. A total of 283 human cases of viral encephalitis with 109 deaths were recorded in Malaysia from 29 September 1998 to December 1999. During the outbreak period, a number of surveillances under three broad groups; Surveillance in Human Health Sector, Surveillance in Animal Health Sector, and Surveillance for the Reservoir Hosts, were carried out to determine the prevalence, risk of virus infections and transmission in human and swine populations as well as the source and reservoir hosts of Nipah virus. Surveillance data showed that the virus spread rapidly among pigs within infected farms and transmission was attributed to direct contact with infective excretions and secretions. The spread of the virus among pig farms within and between states of peninsular Malaysia was due to movement of pigs. The transmission of the virus to humans was through close contact with infected pigs. Human to human transmission was considered a rare event though the Nipah virus could be isolated from saliva, urine, nasal and pharyngeal secretions of patients. Field investigations identified fruitbats of the Pteropid species as the natural reservoir hosts of the viruses. The outbreak was effectively brought under control following the discovery of the virus and institution of correct control measures through a combined effort of multi-ministerial and multidisciplinary teams working in close co-operation and collaboration with other international agencies.


title: Curbing Misinformation and Disinformation in the COVID-19 Era: A View from Cuba.

text: As the COVID-19 health crisis engulfs the planet, we are sub-merged in a parallel pandemic: the glut of misinformation and disinformation. The data associated with this phenomenon are creating a disaster within a disaster. In early April 2020, the Span-ish news agency EFE[1] reported that over one million internet ac-counts were dedicated to rumor-mongering, spreading unverifi ed information about the coronavirus. From January through April 13, fact-checkers at Maldita.es[2] had tracked over 400 lies and false alerts circulated about COVID-19 in Spain alone.


title: The leader RNA of coronavirus mouse hepatitis virus contains an enhancer-like element for subgenomic mRNA transcription.

text: While the 5' cis-acting sequence of mouse hepatitis virus (MHV) for genomic RNA replication has been determined in several defective interfering (DI) RNA systems, it remains elusive for subgenomic RNA transcription. Previous studies have shown that the leader RNA in the DI genome significantly enhances the efficiency of DI subgenomic mRNA transcription, indicating that the leader RNA is a cis-acting sequence for mRNA transcription. To further characterize the cis-acting sequence, we made a series of deletion mutants, all but one of which have an additional deletion of the cis-acting signal for replication in the 5' untranslated region. This deletion effectively eliminated the replication of the DI-chloramphenicol acetyltransferase (CAT)-reporter, as demonstrated by the sensitive reverse transcription (RT)-PCR. The ability of these replication-minus mutants to transcribe subgenomic mRNAs was then assessed using the DI RNA-CAT reporter system. Results from both CAT activity and mRNA transcripts detected by RT-PCR showed that a 5'-proximal sequence of 35 nucleotides (nt) at nt 25 to 59 is a cis-acting sequence required for subgenomic RNA transcription, while the consensus repeat sequence of the leader RNA does not have such effect. Analyses of the secondary structure indicate that this 35-nt sequence forms two stem-loops conserved among MHVs. Deletion of this sequence abrogated transcriptional activity and disrupted the predicted stem-loops and overall RNA secondary structure at the 5' untranslated region, suggesting that the secondary structure formed by this 35-nt sequence may facilitate the downstream consensus sequence accessible for the discontinuous RNA transcription. This may provide a mechanism by which the 5' cis-acting sequence regulates subgenomic RNA transcription. The 5'-most 24 nt are not essential for transcription, while the 9 nt immediately downstream of the leader enhances RNA transcription. The sequence between nt 86 and 135 had little effect on transcription. This study thus defines the cis-acting transcription signal at the 5' end of the DI genome.


Inguinal and incisional hernias.


title: Effects of different doses of sufentanil and remifentanil combined with propofol in target-controlled infusion on stress reaction in elderly patients.

text: The current study aimed to observe the effects of sufentanil and remifentanil combined with propofol in target-controlled infusion (TCI) on perioperative stress reaction in elderly patients. A total of 80 elderly patients requiring general anesthesia were recruited. They were divided into four groups (each n=20) according to different target concentrations of remifentanil and sufentanil. These target concentrations were: 4 ng/ml remifentanil + 0.2 ng/ml sufentanil for group I; 3 ng/ml remifentanil + 0.3 ng/ml sufentanil for group II; 2 ng/ml remifentanil + 0.5 ng/ml sufentanil for anesthesia induction and post-intubation 3 ng/ml remifentanil + 0.2 ng/ml sufentanil for anesthesia maintenance for group III; and 5 ng/ml remifentanil for anesthesia induction and post-intubation 4 ng/ml remifentanil for anesthesia maintenance for group IV. Norepinephrine (NE), epinephrine (E) and angiotensin II (Ang II) levels in plasma were measured prior to the induction of anesthesia, as well as at several different time-points following surgery. The numbers of intraoperative severe hemodynamic fluctuation, postoperative eye-opening and extubation time, and post-extubation restlessness and pain scores were recorded. Group IV had a larger circulation fluctuation control number and higher levels of NE, E and Ang II at 3 h after surgery than any other group (P<0.01). Although group IV had shorter postoperative eye-opening and extubation times compared with the other groups (P<0.05), it also had higher restlessness and pain scores (P<0.01). The combined use of sufentanil and remifentanil stabilizes perioperative hemodynamics and reduces stress hormone levels.


Targeted Temperature Management (TTM 2014).


title: Determination of the lifespan of erythrocytes from greyhounds, using an in vitro biotinylation technique.

text: OBJECTIVE To determine the RBC lifespan of Greyhounds, using an in vitro labeling technique. DESIGN RBC from dogs were labeled with NHS-biotin and their disappearance measured over time to determine RBC lifespan. SAMPLE POPULATION 5 Greyhounds that had been vaccinated against distemper, adenovirus 1 and 2 infections, parainfluenza, leptospirosis, parvovirus, and coronavirus infections, Bordetella bronchiseptica infection, and rabies the previous year; 3 sexually intact 14-month-old Beagles served as controls. PROCEDURE After venipuncture for CBC, catheters were inserted in the cephalic vein of each dog. Butorphanol was then administered to achieve mild sedation and analgesia, and glycopyrrolate was administered to ensure maintenance of adequate heart rate during phlebotomy. Dogs were positioned in lateral recumbency; blood was removed via jugular venipuncture, using a standard laboratory donor blood bag containing citrate-phosphate-dextrose solution. Blood was transferred aseptically into sterile polystyrene containers and NHS-biotin was added. After incubation, the labeled RBC were reinfused into the dogs and the blood was allowed to recirculate for 1 hour before the first postinfusion sample was taken. At frequent intervals, blood to be analyzed was taken by jugular venipuncture, and the percentage of labeled cells was determined by flow cytometry. RESULTS The mean RBC lifespan of non-Greyhounds was significantly longer than that of Greyhounds (104.3 +/- 2.2 days vs 53.6 +/- 6.5 days; P = 0.001). A negative linear correlation was also found between age of the Greyhounds and their RBC lifespan (P = 0.01, R2 = 0.91). CONCLUSIONS The shorter RBC lifespan of the Greyhounds may explain the finding of macrocytosis reported in earlier work. The reason for the shorter RBC lifespan in Greyhounds may be caused by differences in Greyhound RBC membrane structure or accelerated RBC removal from the circulation.


title: Respiratory disease in pregnancy.

text: Respiratory disease is common in pregnancy, and becoming more so: asthma, pneumonia and tuberculosis are all increasing in frequency. This review discusses these conditions and their relationship to pregnancy, as well as considering briefly some rarer diseases.


title: Form and function of joint health and safety committees in Ontario acute care hospitals.

text: Even though joint health and safety committees (JHSCs) have been in existence in Ontario workplaces for almost 30 years and healthcare workers are at high risk for workplace injury, there has been little research done related to JHSC functioning in Ontario's healthcare sector. In his recent review of the 2003 outbreak of severe acute respiratory syndrome in Toronto, Justice Archie Campbell found that JHSCs in Ontario hospitals were not fulfilling their intended roles and responsibilities. The objective of this study was to gain a deeper understanding of the role, resources, structure and functioning of JHSCs in Ontario hospitals.


title: The costs and consequences of methicillin-resistant Staphylococcus aureus infection treatments in Canada.

text: BACKGROUND A multinational randomized controlled trial has shown a trend toward early discharge of patients taking oral linezolid versus intravenous vancomycin (IV) in the treatment of methicillinresistant Staphylococcus aureus (MRSA) infections. Infection treatments resulting in shorter hospitalization durations are associated with cost savings from the hospital perspective. OBJECTIVE To determine whether similar economic advantages are associated with oral linezolid, the costs and consequences of linezolid use following vancomycin IV versus the existing practice in the treatment of infections caused by MRSA were compared. METHODS The charts of all patients admitted to one of three tertiary care teaching hospitals between January 1, 1997 and August 31, 2000 and treated with vancomycin IV for an active MRSA infection (skin and soft tissue only) were reviewed. Based on the vancomycin IV chart review data set and a simulated linezolid data set, the clinical consequences and the associated costs of MRSA treatment with vancomycin IV, and oral and IV forms of linezolid were quantified and compared within the framework of a cost-consequence analysis. RESULTS Patients treated with oral and IV forms of linezolid compared with the existing practice had a shorter length of stay and required fewer home IV care services, which resulted in a cost savings of $750 (2001 values) to the Canadian health care perspective. CONCLUSIONS The estimated cost savings associated with linezolid use not only offset the higher acquisition cost of the anti-infective, but may be substantial to health care systems across Canada, especially as the incidence of MRSA continues to rise.


title: Modified critical care and treatment space considerations for mass casualty critical illness and injury.

text: Mass critical care events are increasingly likely, yet the resource challenges to augment everyday, unrestricted critical care for a surge of disaster victims are insurmountable for nearly all communities. In light of these limitations, an expert panel defined a circumscribed set of key critical care interventions that they believed could be offered to many additional people and yet would also continue to offer substantial life-sustaining benefits for nonmoribund critically ill and injured people. They proposed Emergency Mass Critical Care, which is based on the set of key interventions and includes recommendations for necessary surge medical equipment, treatment space characteristics, and staffing competencies for mass critical care response. To date, Emergency Mass Critical Care is untested, and the real benefits of implementation remain uncertain. Nonetheless, Emergency Mass Critical Care currently remains the only comprehensive construct for mass critical care preparedness and response. This paper reviews current concepts to provide life-sustaining care for hundreds or thousands of people outside of traditional critical care sites.


title: COVID-19: Preliminary Clinical Guidelines for Ophthalmology Practices.

text: The zoonotic Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and its resultant human coronavirus disease (COVID-19) recently appeared as a global health threat that can cause severe respiratory infection and terminal respiratory distress. By the first week of April, more than 1.3 million people had been globally infected and more than 70,000 had lost their lives to this contagious virus. Clinical manifestations occur shortly after exposure, or a few days later. There is controversy regarding the transmission of the virus through the tear and conjunctiva; however, there are reports that the ocular surface might be a potential target for COVID-19. The ease of transmission of this virus at close proximity presents a risk to eyecare workers. Several recommendations have been issued by local and national organizations to address the issue of safe ophthalmic practice during the ongoing COVID-19 pandemic. These guidelines have numerous similarities; however, subtle differences exist. The purpose of this paper was to discuss measures, with a specific focus on standard precautions, to prevent further dissemination of COVID-19 at Eye Clinics. We have proposed procedures to triage suspected cases of COVID-19, considering emergency conditions.


title: Impact of severe acute respiratory syndrome care on the general health status of healthcare workers in taiwan.

text: BACKGROUND The impact of the outbreak of severe acute respiratory syndrome (SARS) was enormous, but few studies have focused on the infectious and general health status of healthcare workers (HCWs) who treated patients with SARS. DESIGN We prospectively evaluated the general health status of HCWs during the SARS epidemic. The Medical Outcome Study Short-Form 36 Survey was given to all HCWs immediately after caring for patients with SARS and 4 weeks after self-quarantine and off-duty shifts. Tests for detection of SARS coronavirus antibody were performed for HCWs at these 2 time points and for control subjects during the SARS epidemic. SETTING Tertiary care referral center in Taipei, Taiwan. SUBJECTS Ninety SARS-care task force members (SARS HCWs) and 82 control subjects. RESULTS All serum specimens tested negative for SARS antibody. Survey scores for SARS HCWs immediately after care were significantly lower than those for the control group (P<.05 by the t test) in 6 categories. Vitality, social functioning, and mental health immediately after care and vitality and mental health after self-quarantine and off-duty shifts were among the worst subscales. The social functioning, role emotional, and role physical subscales significantly improved after self-quarantine and off-duty shifts (P<.05, by paired t test). The length of contact time (mean number of contact-hours per day) with patients with SARS was associated with some subscales (role emotional, role physical, and mental health) to a mild extent. The total number of contact-hours with symptomatic patients with SARS was a borderline predictor (adjusted R2=0.069; P=.038) of mental health score. CONCLUSIONS The impact of the SARS outbreak on SARS HCWs was significant in many dimensions of general health. The vitality and mental health status of SARS HCWs 1 month after self-quarantine and off-duty shifts remained inferior to those of the control group.


title: Cell cycle dependent nucleolar localization of the coronavirus nucleocapsid protein.

text: The nucleolus is a dynamic sub-nuclear structure which is involved in ribosome subunit biogenesis, modulation of cell growth and response to cell stress. The nucleolar proteome varies particularly with regard to the cell cycle. Viral proteins can localise to the nucleolus and using the coronavirus nucleocapsid (N) protein as a model, the cell cycle dependent trafficking of viral proteins to the nucleolus was investigated. Cell synchronisation studies coupled to live cell confocal microscopy indicated that nucleolar localisation of N protein was greater in the G2/M phase of the cell cycle than at other stages. This result was reinforced when FRAP and FLIP analysis indicated that N protein was more mobile within the nucleoplasm and nucleolus in the G2/M phase of the cell cycle. The data suggested that viral nucleolar proteins can also localise to the nucleolus in a cell cycle dependent manner and this may be related to dynamic trafficking.


title: Nosocomial infection control in regional and provincial hospitals.

text: OBJECTIVES To study the organization and practices in nosocomial infection (NI) control in regional and provincial hospitals. MATERIAL AND METHOD Data were collected by questionnaires answered by chairpersons of infection control committees (ICC) infection control nurses (ICNs) and link nurses in regional and provincial hospitals. RESULTS From April to June 2002, sets of questionnaires were sent to all chair persons of regional and provincial hospitals, 120 infection control nurses and 57 link nurses. Of 92 hospitals, 78.3% of chairpersons and all ICNs and link nurses responded. The ICC were chaired by the directors or deputy directors in 26.4% and by doctors in various specialties in the remaining. Among ICNs, 14.2% had never attended a course in infection control and 62.5% had less than 6 years experience. Lack of support from administrators, budget, co-operation from medical personnel and ICN position were the main problems in the IC program. CONCLUSION Nosocomial infection control in regional and provincial hospitals in Thailand needs more support from administrators and more co-operation from medical personnel.


title: Endovascular treatment of carotico-cavernous fistulas with acrylic glue: a series of nine cases.

text: INTRODUCTION Injuries to the internal carotid artery close to the cavernous sinus may result in a fistulous connection between the artery and the venous sinus. Symptoms include pulsatile tinnitus, intracranial bruit, ophthalmological symptoms, and risk of intracerebral hematoma in cases of cortical venous reflux. Previous treatment strategies have included detachable latex balloons, coils, covered stents, or combinations thereof. Today, detachable latex balloons are phased out or withdrawn from several markets. Acrylic glue is a proven stable material used for embolization of arteriovenous shunts. It is a precise, fast, and cost-effective method of endovascular embolization, and it does not cause artifacts on MRI or MRA. METHODS We treated nine patients suffering from direct fistulas with acrylic glue without any permanent neurological adverse events. RESULTS Four patients were treated with glue embolization of the fistula without occlusion of the parent artery. Five patients with long-lasting symptomatology, large tears in the ICA, and with full collateral cerebral circulation were treated with glue embolization of the fistula and sacrifice of the ICA antero- and retrograde via the ICA and the posterior communicating artery. CONCLUSION We suggest acrylic glue to be added to the panel of embolic materials used to treat CCFs.


Prognosis for South Asian and white patients with heart failure in the United Kingdom: counterintuitive findings on heart failure in South Asians may be artefactual.


title: [Logical Explanation of Medical Condition for Patients with Terminal Cancer at Home-Retrospective Analysis of 145 Cases].

text: Frequently, patients with advanced cancer are suffered by various symptoms at the end of life. Furthermore, sometime experience unexpected loss of consciousness(LOC)and/or respiratory arrest. I examined carefully following neck stiffness, LOC, masked face, urinary retention, tonic and/or clonic convulsions, apnea, dysphagia, head rotation, neurogenic pain, hyperthermia, mydriasis 145 patients with advanced cancer. By assuming meningeal carcinomatosis, multifocal neurological findings are logically expected. As a result, before the LOC or respiratory arrest, I was able to logically explain to almost all families of patients, who with suspected neurological signs, expected multifocal neurological signs including LOC or respiratory arrest.


title: Selective infection of lower respiratory tract by respiratory viruses in children with recurrent respiratory tract infections.

text: Thirty preschool children presenting with recurrent respiratory infections and their unaffected siblings were observed prospectively for a year. The index children experienced more episodes of acute respiratory infection than their siblings. Respiratory viruses were the major cause of respiratory infections. The index children had lower respiratory tract disease, predominantly wheeze, during 34% of proved respiratory virus infections compared with 11% of such infections experienced by the control children (p less than 0.02). Atopic children had an increased tendency to wheeze that did not reach significance, but atopy was not associated with increased susceptibility to respiratory infections.


title: Interference of coronavirus infection by expression of IgG or IgA virus neutralizing antibodies.

text: Mouse immunoglobulin gene fragments encoding the variable modules of the heavy (VH) and light (VL) chains of a transmissible gastroenteritis coronavirus (TGEV) neutralizing monoclonal antibody (MAb) have been cloned and sequenced. The selected MAb recognizes a highly conserved viral epitope and does not lead to the selection of neutralization escape mutants. Chimeric immunoglobulin genes with the variable modules from the murine MAb and constant modules of human gamma 1 and kappa chains were constructed using RT-PCR. These chimeric immunoglobulins were stably or transiently expressed in murine myelomas and COS cells, respectively. The secreted recombinant antibodies had radioimmunoassay (RIA) titers higher than 10(3) and reduced the infectious virus more than 10(4)-fold. Recombinant dimeric IgA showed a 50-fold enhanced neutralization of TGEV relative to a recombinant monomeric IgG1 which contained the identical antigen binding site. Epithelial cell lines stably-transformed with these constructs and expressing either recombinant IgG or IgA TGEV neutralizing antibodies reduced virus production by > 10(5)-fold after infection with homologous virus, although a residual level of virus production (< 10(2) PFU/ml) remained in less than 0.1% of the cells.


title: Characterization of determinants involved in the feline infectious peritonitis virus receptor function of feline aminopeptidase N.

text: Feline aminopeptidase N (fAPN) is a major cell surface receptor for feline infectious peritonitis virus (FIPV), transmissible gastroenteritis virus (TGEV), human coronavirus 229E (HCV 229E) and canine coronavirus (CCV). By using chimeric molecules assembled from porcine, human and feline APN we have analysed the determinants involved in the coronavirus receptor function of fAPN. Our results show that amino acids 670-840 of fAPN are critically involved in its FIPV and TGEV receptor function whereas amino acids 135-297 are essential for the HCV 229E receptor function. We also demonstrate that a chimeric molecule assembled from human and porcine APN is able to act as a receptor for FIPV. This is surprising as neither human nor porcine APN by themselves mediate FIPV infection. These results suggest that different determinants in the APN protein are involved in mediating the coronavirus receptor function.


title: A coding RNA sequence acts as a replication signal in cardioviruses.

text: Theiler's virus and Mengo virus are representatives of the Cardiovirus genus within the picornavirus family. Their genome is an 8-kilobase long positive strand RNA molecule. This RNA molecule plays three roles in infected cells: It serves as a messenger RNA, acts as a template for genome replication, and is encapsidated to form progeny virions. We observed that a cis-acting signal required for replication of Theiler's virus was contained within a 130-nt stretch of the region encoding the capsid protein VP2. This RNA sequence does not influence internal ribosome entry site-mediated translation initiation and thus likely acts directly as a signal for the replication complex. We found a similar signal in the VP2-coding sequence of Mengo virus, and both signals could be functionally exchanged. Within the replication element, a 9-nt sequence that is highly conserved among cardioviruses was shown to be essential for replication. This conserved sequence was contained in mostly unpaired regions of the RNA secondary structure predicted for the replication elements of the various cardioviruses. Interestingly, a similar replication element has been reported to occur in the distantly related human rhinovirus type 14, suggesting that such elements could be conserved throughout the picornavirus family. However, the different location of the replication elements in rhinovirus and cardioviruses, and the fact that they were not functionally exchangeable, is raising intriguing questions about the evolution of such signals in picornaviruses.


title: How will Australian general practitioners respond to an influenza pandemic? A qualitative study of ethical values.

text: OBJECTIVES To explore general practitioners' perceptions of their preparedness for an influenza pandemic, the changes they would make to their practice, and the ethical justifications for their planned actions. DESIGN AND SETTING A qualitative study was performed among South Australian GPs between March and October 2007. A semi-structured interview was carried out with each participant in his or her practice, and the interviews were audio-recorded, transcribed and analysed thematically. PARTICIPANTS 10 GPs were recruited: five from a metropolitan Division and five from a rural Division of General Practice. RESULTS Some participants felt they would not be able to cope with an influenza pandemic, while others felt it would simply mean an increase in their workloads. Most respondents considered creating separate waiting rooms, moving the reception desk outside of the practice and delaying all non-urgent consultations in order to deal with a pandemic more effectively. Respondents mentioned the conflict between their various roles and responsibilities as a primary source of tension when thinking about the way they would organise their work in the event of a pandemic. A number of GPs said they would not practise in the event of a pandemic, as they felt their responsibility to their families outweighed that to their patients. CONCLUSIONS Professional codes of ethics should include guidance about the scope of the duty to treat during infectious disease outbreaks. The community has to uphold the value of reciprocity, and ensure that GPs and their families are provided with support during a pandemic and are given the opportunity to be actively involved in pandemic preparedness planning.


title: Comparison of complement fixation with two enzyme-linked immunosorbent assays for the detection of antibodies to respiratory viral antigens.

text: We compared complement fixation (CF) for the measurement of antibodies against influenza A, influenza B, respiratory syncytial virus (RSV), human adenovirus, and parainfluenza viruses 1, 2, and 3 (para-1, para-2, and para-3) with 2 enzyme-linked immunosorbent assays (ELISA kits, A and B). The IgG ELISA kits compared very well with each other except for the influenza A and B IgG ELISAs. The IgG ELISAs, in general, did not agree with CF In contrast, the IgM ELISAs compared well with CF and each other except for the consensus parainfluenza panel from ELISA B. The poor agreement of the IgG ELISAs with the CF test can be explained by the increased sensitivity of the ELISAs and differences between CF antigens and the ELISA antigens. The influenza A and influenza B ELISA antigens differed between both kits, which may explain their poor agreement. The ELISA is a suitable replacement for CF, providing greater sensitivity, isotype specificity, and ease of use.


COVID-19 in hem/onc news.


title: The effect of doxorubicin loading on response and toxicity with drug-eluting embolization in resectable hepatoma: a dose escalation study.

text: AIM The dose-response relationship between doxorubicin and superabsorbent drug-eluting microspheres has not been established. In this study, we investigated the relationships between dose and delivery parameters as they pertain to toxicity and response in surgically resectable hepatocellular carcinoma (HCC). PATIENTS AND METHODS Twenty-five patients with resectable HCC were randomly assigned and divided into four groups, each receiving either bland, 25 mg, 50 mg or 75 mg of doxorubicin loaded Super Absorbent Polymer microspheres, with 24 patients undergoing surgical resection. Response Evaluation and Criteria in Solid Tumors (RECIST) 1.0 and European Association for the Study of the Liver (EASL)-based volumetric response was performed at one month and surgical resection of the reference tumor was performed at two months. Adverse events were collected at regular intervals. RESULTS Fifty-six percent of patients demonstrated complete response according to EASL criteria as opposed to 0% according to RECIST (v1.0) criteria. Residual tumor was identified in all groups (0 mg: 35%±28.5%; 25 mg: 42%±30.4%; 50 mg: 3.6%±3.3%; and 75 mg: 49.29%±32.6%. A total of 112 adverse events of grades 1-3 occurred (average 5.1 per patient), with no grade 4 or 5. No difference was noted between bland embolic and drug-loaded groups. Subset analysis did demonstrate a significantly increased degree of necrosis in the 50 mg-loaded group (p=0.018). Strong correlation existed between arterial phase Computer Tomography EASL-based response and histopathology (r=0.81; p<0.0001). All groups had residual tumor. CONCLUSION Histology correlates strongly with one-month post-procedural imaging and response optimized at 50 mg of loading per vial. Adverse events were a reflection of embolization, with no relationship between loading dose or administered dose of doxorubicin.


title: Pharmacotherapy Considerations in CKD Patients With COVID-19, A Narrative Review.

text: Treatment of coronavirus disease 2019 (COVID-19) among patients with CKD requires special pharmacotherapy considerations that are reviewed here. Literature review was done for several pharmacotherapy aspects in CKD patients including selection and modification of COVID-19 treatment, drug interactions, nephrotoxicity of drugs that are used for treatment of COVID-19 and potential risks/benefits of routine medications of CKD patients during COVID-19 pandemic. CKD patients should be treated according to local or national COVID-19 protocols as other patients. But, there is no data on using remdesivir in patients with severe CKD. Oseltamivir and ribavirin require dose modification in patients with moderate to severe CKD. Nephrolithiasis, CKD, and acute interstitial nephritis have been reported with protease inhibitors. Acute kidney injury has been reported with remdesivir in patients with severe COVID-19. Pharmacokinetic-enhanced protease inhibitors increase the concentration of some drugs such as statins, cinacalcet, steroids, calcineurin inhibitors (CNIs). Some hypothetical benefits and harms have been suggested for statins and renin-angiotensinaldosterone system inhibitors in COVID-19 patients. Continuing guideline-directed administration of these drugs is recommended. Among different immunomodulating/immunosuppressive drugs, hydroxychloroquine and CNIs are the safest ones during COVID-19. Antimetabolites are suggested to be withheld during moderate to severe COVID-19. Fluid therapy and anticoagulant prophylaxis/ treatment need special attention in CKD patients with COVID-19. CKD patients with COVID-19 are treated as other patients, with some dose modifications if needed. Be mindful for management of drug interactions as well as modification of immunosuppressive drugs in patients with moderate to severe COVID-19.


title: Norwalk virus RNA is infectious in mammalian cells.

text: Human noroviruses are positive-sense RNA viruses and are the leading cause of epidemic acute viral gastroenteritis in developed countries. The absence of an in vitro cell culture model for human norovirus infection has limited the development of effective antivirals and vaccines. Human histo-blood group antigens have been regarded as receptors for norovirus infection, and expression of the alpha(1,2) fucosyltransferase gene (FUT2) responsible for the secretor phenotype is required for susceptibility to Norwalk virus (NV) infection. We report for the first time that transfection of NV RNA, isolated from stool samples from human volunteers, into human hepatoma Huh-7 cells leads to viral replication, with expression of viral antigens, RNA replication, and release of viral particles into the medium. Prior treatment of the RNA with proteinase K completely abolishes RNA infectivity, suggesting a key role of an RNA-protein complex. Although overexpression of the human FUT2 gene enhances virus binding to cells, it is not sufficient to allow a complete viral infection, and viral spread from NV-transfected cells to naïve cells does not occur. Finally, no differences in NV RNA replication are observed between Huh-7 and Huh-7.5.1 cells, which contain an inactivating mutation in retinoic acid-inducible gene I (RIG-I), suggesting that the RIG-I pathway does not play a role in limiting NV replication. Our results strongly suggest that the block(s) to NV replication in vitro is at the stage of receptor and/or coreceptor binding and/or uncoating, either because cells lack some specific factor or activation of cellular antiviral responses independent of RIG-I inhibits virus replication.


title: The UCUAAAC promoter motif is not required for high-frequency leader recombination in bovine coronavirus defective interfering RNA.

text: The 65-nucleotide leader on the cloned bovine coronavirus defective interfering (DI) RNA, when marked by mutations, has been shown to rapidly convert to the wild-type leader of the helper virus following DI RNA transfection into helper virus-infected cells. A model of leader-primed transcription in which free leader supplied in trans by the helper virus interacts by way of its flanking 5'UCUAAAC3' sequence element with the 3'-proximal 3'AGAUUUG5' promoter on the DI RNA minus strand to prime RNA replication has been used to explain this phenomenon. To test this model, the UCUAAAC element which occurs only once in the BCV 5' untranslated region was either deleted or completely substituted in input DI RNA template, and evidence of leader conversion was sought. In both cases, leader conversion occurred rapidly, indicating that this element is not required on input RNA for the conversion event. Substitution mutations mapped the crossover region to a 24-nucleotide segment that begins within the UCUAAAC sequence and extends downstream. Although structure probing of the bovine coronavirus 5' untranslated region indicated that the UCUAAAC element is in the loop of a prominent stem and thus theoretically available for base pair-directed priming, no evidence of an unattached leader early in infection that might have served as a primer for transcription was found by RNase protection studies. These results together suggest that leader conversion on the DI RNA 5' terminus is not guided by the UCUAAAC element and might arise instead from a high-frequency, region-specific, homologous recombination event perhaps during minus-strand synthesis rather than by leader priming during plus-strand synthesis.


title: Normothermia is Best Achieved by Warming Above and Below with Pre-warming Adjunct: A Comparison of Conductive Fabric Versus Forced-air and Water.

text: BACKGROUND Although forced-air warming is the most commonly used method for perioperative patient warming, it is fundamentally problematic because it disturbs the carefully designed airflow in the operating room. Because unintended hypothermia has significant consequences, there is a need for more effective warming strategies. The effectiveness of warming technologies that apply heat through the skin is based on surface-area contact with the heat source and the duration of pre-warming. Therefore, we sought to test the therapeutic effectiveness of combined above- and below-warming therapies. Our hospital prohibits forced-air warming before the patient is draped, so a secondary goal was to determine the effect of preoperative warming using a system that does not interfere with airflow in the operating room. METHODS We prospectively randomized 35 patients undergoing total knee arthroplasty into two groups: 1) forced-air warming/water mattress, using both WarmTouch® upper-body forced-air warming (Medtronic/Covidien Inc., Dublin, Ireland) and a Norm-O-Temp® underbody water mattress (CSZ/Gentherm Inc., Cincinnati, OH, USA), and 2) conductive fabric warming, using a HotDog® electric upper-body blanket (Augustine Temperature Management LLC, Eden Prairie, MN, USA) and a HotDog® underbody mattress. RESULTS Throughout the surgical procedure, group 2 patients had significantly higher temperatures; this group experienced superior pre-warming during preoperative preparations and thus the redistribution temperature drop following the induction of anesthesia was reduced. Both groups achieved 100% normothermia by the end of surgery. CONCLUSION Based solely on the temperatures at the end of surgery, these data indicate that forced-air warming in conjunction with a water mattress warming system is as effective as a conductive fabric electric warming system alone.


Reduction of rotavirus-, coronavirus- and E. coli-associated calf-diarrheas in a large-size dairy herd by means of dam vaccination with a triple-vaccine.


title: Natural cytotoxicity against mouse hepatitis virus-infected target cells. I. Correlation of cytotoxicity with virus binding to leukocytes.

text: Spleen cells from uninfected control mice selectively lysed BALB/c 3T3 fibroblasts infected with mouse hepatitis virus (MHV), a murine coronavirus. Lysis of infected cells occurred within 3 hr, and histocompatibility between effector and target cells was not required. This natural, cell-mediated, virus-associated cytotoxicity differed from NK cell- and T cell-mediated lysis. Spleen cells from animals infected with MHV were enriched in NK activity and were more cytotoxic to YAC-1 target cells, but did not show enhanced cytotoxicity for MHV-infected target cells. Spleen cells from beige mice, which are deficient in NK cell activity, were able to lyse MHV-infected target cells, as were spleen cells from nude mice, which are deficient in T cell activity. Lysis of MHV-infected target cells could be mediated by cells from the spleen and, to a lesser extent, by cells from the bone marrow, but not by resident peritoneal cells or thymocytes. We suggest the term "virus killer (VK) activity" for this phenomenon. VK activity of splenocytes from different mouse strains correlated with the ability of the splenocytes to bind purified radiolabeled MHV virions. MHV virions caused agglutination of spleen leukocytes from susceptible mouse strains, indicating that leukocyte agglutination or adsorption may provide a useful assay for coronaviruses such as MHV which lack hemagglutinating activity. SJL mouse splenocytes did not bind MHV and did not lyse infected targets. MHV bound relatively well to splenocytes of other mouse strains, but poorly to thymocytes and erythrocytes. Binding of MHV to leukocytes was not influenced by 6 mM EDTA or EGTA, indicating a lack of requirement for Mg++ or Ca++. VK activity was also resistant to EDTA and EGTA, in contrast to NK activity, which was sensitive to those chelating agents. VK activity was also unaffected by actinomycin D, cycloheximide, or puromycin, indicating that new protein synthesis was not required for lysis. Antibody to interferon-alpha/beta did not block lysis, nor was there substantially enhanced lysis mediated by leukocytes from mice infected with virus and thus exposed to high levels of interferon. VK activity was blocked by antibody directed against the peplomeric glycoprotein E2 of MHV. VK activity required infected target cells, because cells with adsorbed MHV virions were not lysed by splenocytes.(ABSTRACT TRUNCATED AT 400 WORDS)


title: IP-10 Is Elevated in Virus-Induced Acute Exacerbations in Childhood Asthma.

text: OBJECTIVE Viral infections and sensitization to aeroallergens are major factors in the exacerbation of asthma and its development during early childhood. However, the cytokine profiles and eosinophil activation status linked to the association between viral infections and sensitization to aeroallergens are incompletely understood. Here we investigated respiratory viruses, serum eosinophil cationic protein (ECP), and various cytokines/chemokines in acute exacerbation of childhood asthma. METHODS We analyzed peripheral eosinophil counts, serum ECP, and 27 cytokines/chemokines in 76 virus-induced acute asthma cases with or without aeroallergen sensitization. Asthma due to sensitization was defined by a positive reaction to at least one aeroallergen in serum specific IgE antibody tests. Virus detection was performed using antigen detection kits and/or RT-PCR, followed by direct DNA sequencing analysis. Serum cytokines/chemokines were measured using a multi-cytokine detection system. RESULTS Peripheral eosinophil counts and serum ECP and IL-5 levels were significantly elevated in sensitized cases compared with nonsensitized cases. Conversely, IP-10 values were significantly higher in nonsensitized cases. An inverse correlation between IP-10 and IL-5 production was identified in virus-induced acute exacerbations of asthma but not in controls. CONCLUSIONS Cytokine profiles and eosinophil activation status might be different between sensitized and nonsensitized cases of virus-induced acute exacerbations of asthma.