title: Bowlby and Robertson revisited: the impact of isolation on hospitalized children during SARS.

text: Psychosocial issues related to pediatric isolation remain relevant to both clinical practice and research. The early theories of Bowlby and Robertson are central to this discussion for it was predominantly their work that elicited several policy changes aimed at improving psychosocial care in pediatric settings. Recognizing the significant effects of isolating hospitalized children is just as relevant today where serious infectious diseases can lead to separation of family members. An ethnographic, qualitative study examined the experiences and perspectives of children hospitalized because of SARS (severe acute respiratory syndrome), their parents, and pediatric health care providers. The sample included 23 participants: five children, 10 parents, and eight health care providers. Data analyses illuminated a range of perceived experiences for this triadic sample. Themes related to the effects of social isolation on children were predominant. They included emotional upheaval, changes in parental and professional roles, and familial experiences following hospital discharge. Finally, the paper draws on the findings of the present study and other recent work in offering recommendations for effective clinical approaches in the event of future outbreaks.


title: Antibodies as delivery vehicles for radioimmunotherapy of infectious diseases.

text: The field of infectious diseases is in crisis and there is a need for strategies that can facilitate the rapid development of new antimicrobial agents. Radioimmunotherapy (RIT), a therapeutic modality originally developed for cancer treatment, has recently been suggested as a novel therapy for the treatment of a variety of infectious diseases. Because specific antibodies are used in RIT as delivery vehicles of cytocidal radiation, their molecular weight influences the nonspecific accumulation in infectious foci and blood clearance, and their affinity-specific accumulation of antibodies in infectious foci. Like the problems encountered in oncology, relevant variables in the development of RIT of infectious diseases include target antigen-shedding; delivering radionuclides to infectious foci in organs, abscesses, granulomas, heart and brain, and potential safety concerns. Dadachova and Casadevall anticipate that RIT can be developed for many types of infectious diseases, including microbes resistant to conventional antimicrobial therapy and agents of biological warfare.


title: Innovation in veterinary medical education: the concept of 'One World, One Health' in the curriculum of the Faculty of Veterinary Medicine at the University of Calgary.

text: 'One World, One Health' is a foundation concept in veterinary medicine, much like comparative medicine. However, teachers of veterinary medicine often fail to identify it or speak of its importance within the veterinary curriculum. The resurgence of interest in the 'One World, One Health' concept aligns well with the underlying principles on which the University of Calgary Faculty of Veterinary Medicine (UCVM) has been newly founded. This concept is therefore a key component of the UCVM programme, and one that is well highlighted for those studying in the Doctor of Veterinary Medicine (DVM) course and graduate students.


title: Crystal structure of aminopeptidase N (proteobacteria alanyl aminopeptidase) from Escherichia coli and conformational change of methionine 260 involved in substrate recognition.

text: Aminopeptidase N from Escherichia coli is a broad specificity zinc exopeptidase belonging to aminopeptidase clan MA, family M1. The structures of the ligand-free form and the enzyme-bestatin complex were determined at 1.5- and 1.6-A resolution, respectively. The enzyme is composed of four domains: an N-terminal beta-domain (Met(1)-Asp(193)), a catalytic domain (Phe(194)-Gly(444)), a middle beta-domain (Thr(445)-Trp(546)), and a C-terminal alpha-domain (Ser(547)-Ala(870)). The structure of the catalytic domain exhibits similarity to thermolysin, and a metal-binding motif (HEXXHX(18)E) is found in the domain. The zinc ion is coordinated by His(297), His(301), Glu(320), and a water molecule. The groove on the catalytic domain that contains the active site is covered by the C-terminal alpha-domain, and a large cavity is formed inside the protein. However, there exists a small hole at the center of the C-terminal alpha-domain. The N terminus of bestatin is recognized by Glu(121) and Glu(264), which are located in the N-terminal and catalytic domains, respectively. Glu(298) and Tyr(381), located near the zinc ion, are considered to be involved in peptide cleavage. A difference revealed between the ligand-free form and the enzyme-bestatin complex indicated that Met(260) functions as a cushion to accept substrates with different N-terminal residue sizes, resulting in the broad substrate specificity of this enzyme.


title: Comparison of NxTAG Respiratory Pathogen Panel and Anyplex II RV16 Tests for Multiplex Detection of Respiratory Pathogens in Hospitalized Children.

text: Multiplex molecular techniques can detect a diversity of respiratory viruses and bacteria that cause childhood acute respiratory infection rapidly and conveniently. However, currently available techniques show high variation in performance. We sought to compare the diagnostic accuracy of the novel multiplex NxTAG respiratory pathogen panel (RPP) RUO test versus a routine multiplex Anyplex II RV16 assay in respiratory specimens collected from children <18 years of age hospitalized with nonspecific symptoms of acute lower respiratory infection. Parallel testing was performed on nasopharyngeal aspirates prospectively collected at referral Children's Hospital Sant Joan de Déu (Barcelona, Spain) between June and November 2015. Agreement values between the two tests and kappa coefficients were assessed. Bidirectional sequencing was performed for the resolution of discordant results. A total of 319 samples were analyzed by both techniques. A total of 268 (84.0%) of them yielded concordant results. Positive percent agreement values ranged from 83.3 to 100%, while the negative percent agreement was more than 99% for all targets except for enterovirus/rhinovirus (EV/RV; 94.4%). Kappa coefficients ranged from 0.83 to 1.00. Discrepancy analysis confirmed 66.0% of NxTAG RPP RUO results. A total of 260 viruses were detected, with EV/RV (n = 105, 40.4%) being the most prevalent target. Viral coinfections were found in 44 (14.2%) samples. In addition, NxTAG RPP RUO detected single bacterial and mixed viral-bacterial infections in seven samples. NxTAG RPP RUO showed high positive and negative agreement with Anyplex II RV16 for main viruses that cause acute respiratory infections in children, coupled with an additional capability to detect some respiratory bacteria.


title: Coronavirus MHV-A59 causes upregulation of interferon-beta RNA in primary glial cell cultures.

text: Infection of mice with coronavirus mouse hepatitis virus strain MHV-A59 causes focal acute encephalitis, hepatitis and chronic demyelinating disease. To investigate host interferon (IFN) response to viral infection within the brain, RNA was extracted from A59-or MHV-2- infected and mock-infected primary astrocyte cultures from newborn mice, RT-PCR amplified RNA with primers specific for the various IFNs, transferred to nylon membranes and hybridized with IFN specific digoxigenin-labeled probes. Infection of primary astrocyte cultures from newborn mice with either A59 or MHV-2 caused upregulation of IFN-beta RNA, but not IFN-gamma or IFN-alpha. Thus, brain astrocytes are capable of producing a local IFN-beta response upon infection with MHV. The response of the other IFNs following MHV infection may be derived from inflammatory cells.


title: Clearance of infection in cats naturally infected with feline coronaviruses is associated with an anti-S glycoprotein antibody response.

text: We have investigated by Western blotting the antibody responses against the three major structural proteins in cats naturally infected with feline coronaviruses that cleared virus infection (group I), established chronic asymptomatic infection (group II) or were sick (group III). The cats of group I developed an anti-S glycoprotein response that was, relative to the anti-M glycoprotein response, at least 30-fold higher than that of chronically infected cats from groups II and III. These results suggest that the anti-S glycoprotein response against antigenic domains revealed by Western blot is associated with clearance of the virus after natural infection, and is not a risk factor for the establishment of a chronic infection.


title: Seek local advice to control SARS.

text: NURSES ARE being urged to practise basic infection control and make sure they have a thorough knowledge of their hospital's own procedures to combat any out- break of the Severe Acute Respiratory Syndrome (SARS) virus.


title: Antibody dependent cellular cytotoxicity against coronavirus 229E-infected cells.

text: An antibody dependent cellular cytotoxic (ADCC) reaction was elicited using human mixed lymphocyte cultures against coronavirus 229E-infected C-16 cells. The response was assayed in microtitre plates by radioactive chromium release. Optimal killing of target cells was achieved using cells labelled 22 h after infection. A prozone was present and the optimal dilution of sera was invariably 1:200. The majority (70%) of sera tested gave a positive ADCC response. These included all sera judged positive by ELISA, but also some which were judged negative. Two experiments using purified IgG from serum which was ADCC and ELISA positive suggested that the response is mediated by an IgG antibody and that the prozone is due to a separate serum component.


title: Rapid Reduction in Use of Antidiabetic Medication after Laparoscopic Sleeve Gastrectomy: The Newfoundland and Labrador Bariatric Surgery Cohort (BaSCo) Study.

text: BACKGROUND Patients who have undergone bariatric surgery generally need fewer medications as they experience improvement in, or even resolution of, various medical conditions, including type 2 diabetes mellitus, hypertension, and dyslipidemia. Published data on changes in medication use after laparoscopic sleeve gastrectomy, a type of bariatric surgery that is growing in popularity, are limited. OBJECTIVE To determine whether patients took fewer medications for management of type 2 diabetes, hypertension, and dyslipidemia after laparoscopic sleeve gastrectomy, relative to preprocedure medications. METHODS In this prospective, single-centre cohort study, a nurse practitioner used standard medication reconciliation and study data-extraction forms to interview adult patients who had undergone laparoscopic sleeve gastrectomy and determine their medication use and pertinent demographic data. The data were analyzed using generalized estimating equations and standard statistical software. Outcome measures included changes in the use of antidiabetic, antihypertensive, and antilipemic medications at 1, 3, and 6 months after the surgery. RESULTS A total of 65 patients who underwent laparoscopic sleeve gastrectomy between May 2011 and January 2014 met the study inclusion criteria. Before surgery, the 30 patients with type 2 diabetes were taking an average of 1.9 antidiabetic medications. One month after the procedure, 15 (50%) had discontinued all antidiabetic medications, with a further decline at 3 and 6 months (p < 0.001 at each time point). Among the patients who were taking antihypertensives (n = 48) and antilipemics (n = 33) before surgery, the decline in use occurred at a more modest rate, with 6 (12%) and 2 (6%), respectively, discontinuing these medication classes within 1 month, and 12 (25%) (p = 0.001) and 8 (24%) (p = 0.015) having discontinued by 6 months. CONCLUSIONS These findings suggest that patients with a history of type 2 diabetes mellitus, hypertension, and/or dyslipidemia who undergo laparoscopic sleeve gastrectomy are less likely to require disease-specific medications shortly after surgery.


SARS : A Tale of Two Epidemics.


Coronavirus infections in military recruits. Three-year study with coronavirus strains OC43 and 229E.


Evidence of close relatedness between turkey and bovine coronaviruses.


title: Bovine coronavirus uses N-acetyl-9-O-acetylneuraminic acid as a receptor determinant to initiate the infection of cultured cells.

text: The importance of N-acetyl-9-O-acetylneuraminic acid (Neu5,9Ac2) as a receptor determinant for bovine coronavirus (BCV) on cultured cells was analysed. Pretreatment of MDCK I (Madin Darby canine kidney) cells with neuraminidase or acetylesterase rendered the cells resistant to infection by BCV. The receptors on a human (CaCo-2) and a porcine (LLC-PK1) epithelial cell line were also found to be sensitive to neuraminidase treatment. The susceptibility to infection by BCV was restored after resialylation of asialo-MDCK I cells with Neu5,9Ac2. Transfer of sialic acid lacking a 9-O-acetyl group was ineffective in this respect. These results demonstrate that 9-O-acetylated sialic acid is used as a receptor determinant by BCV to infect cultured cells. The possibility is discussed that the initiation of a BCV infection involves the recognition of different types of receptors, a first receptor for primary attachment and a second receptor to mediate the fusion between the viral envelope and the cellular membrane.


title: Cross-neutralization of human and palm civet severe acute respiratory syndrome coronaviruses by antibodies targeting the receptor-binding domain of spike protein.

text: The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) is considered as a protective Ag for vaccine design. We previously demonstrated that the receptor-binding domain (RBD) of S protein contains multiple conformational epitopes (Conf I-VI) that confer the major target of neutralizing Abs. Here we show that the recombinant RBDs derived from the S protein sequences of Tor2, GD03, and SZ3, the representative strains of human 2002-2003 and 2003-2004 SARS-CoV and palm civet SARS-CoV, respectively, induce in the immunized mice and rabbits high titers of cross-neutralizing Abs against pseudoviruses expressing S proteins of Tor2, GD03, and SZ3. We also demonstrate that the Tor2-RBD induced-Conf I-VI mAbs can potently neutralize both human SARS-CoV strains, Tor2 and GD03. However, only the Conf IV-VI, but not Conf I-III mAbs, neutralize civet SARS-CoV strain SZ3. All these mAbs reacted significantly with each of the three RBD variants (Tor2-RBD, GD03-RBD, and SZ3-RBD) that differ at several amino acids. Regardless, the Conf I-IV and VI epitopes were completely disrupted by single-point mutation of the conserved residues in the RBD (e.g., D429A, R441A, or D454A) and the Conf III epitope was significantly affected by E452A or D463A substitution. Interestingly, the Conf V epitope, which may overlap the receptor-binding motif and induce most potent neutralizing Abs, was conserved in these mutants. These data suggest that the major neutralizing epitopes of SARS-CoV have been apparently maintained during cross-species transmission, and that RBD-based vaccines may induce broad protection against both human and animal SARS-CoV variants.


Incidentally Diagnosed COVID-19 Infection in Trauma Patients; a Clinical Experience.


title: The N-terminal region of the murine coronavirus spike glycoprotein is associated with the extended host range of viruses from persistently infected murine cells.

text: Although murine coronaviruses naturally infect only mice, several virus variants derived from persistently infected murine cell cultures have an extended host range. The mouse hepatitis virus (MHV) variant MHV/BHK can infect hamster, rat, cat, dog, monkey, and human cell lines but not the swine testis (ST) porcine cell line (J. H. Schickli, B. D. Zelus, D. E. Wentworth, S. G. Sawicki, and K. V. Holmes, J. Virol. 71:9499-9507, 1997). The spike (S) gene of MHV/BHK had 63 point mutations and a 21-bp insert that encoded 56 amino acid substitutions and a 7-amino-acid insert compared to the parental MHV strain A59. Recombinant viruses between MHV-A59 and MHV/BHK were selected in hamster cells. All of the recombinants retained 21 amino acid substitutions and a 7-amino-acid insert found in the N-terminal region of S of MHV/BHK, suggesting that these residues were responsible for the extended host range of MHV/BHK. Flow cytometry showed that MHV-A59 bound only to cells that expressed the murine glycoprotein receptor CEACAM1a. In contrast, MHV/BHK and a recombinant virus, k6c, with the 21 amino acid substitutions and 7-amino-acid insert in S bound to hamster (BHK) and ST cells as well as murine cells. Thus, 21 amino acid substitutions and a 7-amino-acid insert in the N-terminal region of the S glycoprotein of MHV/BHK confer the ability to bind and in some cases infect cells of nonmurine species.


American Association of Clinical Endocrinologists and American Association of Endocrine Surgeons Medical Guidelines for the Management of Adrenal Incidentalomas: executive summary of recommendations.


title: [Response plan in the neonatal intensive care unit during epidemic of SARS-CoV-2 infection (2nd Edition)].

text: Since December 2019, an outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has spread from China to other countries. In order to effectively respond to possible neonatal SARS-CoV-2 infection, neonatologists from the Medical Association of Chinese People's Liberation Army and the Editorial Committee of Chinese Journal of Contemporary Pediatrics proposed the response plan in the neonatal intensive care unit during epidemic of SARS-CoV-2 infection (1st edition) at the end of January of 2020. Based on the further knowledge and experience on SARS-CoV-2 infection, the neonatologists updated the plan according to the current evidence, so as to provide a better guide for clinical medical staff to deal with the SARS-CoV-2 infection in the NICU.


SARS triggers biomedical shake-up in China.


title: [Severe inflammation in children - cause for awareness during the current pandemic].

text: Clinical manifestations of COVID-19 in children are generally mild and many are asymptomatic. However, recent international reports have described clusters of children presenting with an unusual syndrome of high, persistent fever, severe inflammation and single- or multiorgan dysfunction, with a suspected association to COVID-19 pandemic. It is recommended that clinicians are aware of this rare syndrome and that suspected cases are treated in a multidisciplinary setting.


title: Challenges of Covid-19 testing.

text: No abstract available.


title: Strategies to reduce person-to-person transmission during widespread Escherichia coli O157:H7 outbreak.

text: During the Escherichia coli O157:H7 outbreak in 2006 in the United States, the primary strategy to prevent illness was to advise consumers not to eat spinach. No widespread warnings were issued about preventing person-to-person (secondary) transmission. A disease transmission model, fitted to the current data, was used to investigate likely reductions in illnesses that could result from interventions to prevent secondary transmission. The model indicates that exposure to contaminated spinach occurred early in the outbreak and that secondary transmission was similar to that in previous E. coli outbreaks ( 12%). The model also suggests that even a modestly effective strategy to interrupt secondary transmission (prevention of only 2%-3% of secondary illnesses) could result in a reduction of 5%-11% of symptomatic cases. This analysis supports the use of widespread public health messages during outbreaks of E. coli O157:H7 with specific advice on how to interrupt secondary transmission.


The vaccine crossroads.


title: The seXY side of COVID-19: what is behind female protection?

text: Not availble.


title: Health status and access to health care of migrant workers in China.

text: OBJECTIVES We explored living and working conditions, health status, and health-care access in Chinese rural-to-urban migrants and compared them with permanent rural and urban residents. METHODS A questionnaire was administered to 1,958 urban workers, 1,909 rural workers, and 4,452 migrant workers in Zhejiang Province, Eastern China, in 2004. Blood samples for human immunodeficiency virus (HIV) and syphilis were taken from the migrant and urban workers. RESULTS Migrants were young, worked very long hours, and their living conditions were very basic. Nineteen percent had some form of health insurance and 26% were entitled to limited sick pay compared with 68% and 66%, respectively, for urban workers. Migrants had the best self-rated health and reported the least acute illness, chronic disease, and disability, after controlling for age and education. There were no HIV infections detected in either the migrant or urban workers. However, 15 urban workers (0.68%, 95% confidence interval [CI] 0.35, 1.02) and 20 migrants (0.48%, 95% CI 0.26, 0.66, p=0.06) tested positive for syphilis. The high cost of health care in the city was a barrier to health-care access in the last year for 15% of the migrants and 8% of the urban workers. Forty-seven percent of the migrants were unwilling to make contributions to health insurance. CONCLUSIONS These migrants demonstrated the "healthy migrant effect." However, poor living conditions and inattention to health may make migrants vulnerable to poor long-term health. Because health insurance schemes will remain limited for the forseeable future, attention should focus on providing affordable health care to both uninsured migrants and the urban poor.


Coronavirus SD-induced immunoregulatory disturbances in a murine model of demyelination.


title: [Laparoscopic myomectomy and use of electromechanical morcellator: clinical results in a series of cases].

text: OBJECTIVE to describe the clinical and operative characteristics of patients undergoing laparoscopic myomectomy and use of electromechanical morcellator for removal of tissue. MATERIAL AND METHODS An observational, descriptive and retrospective study was done with the medical records of 65 patients that underwent laparoscopic myomectomy from July 2008 to July 2013. The clinical outcomes, including body mass index (BMI), pregnancies number, parity, previous abortions, indication of myomectomy, surgical findings, length of the procedure, operative bleeding, entry into the endometrial cavity, operative complications and conversion rate to laparotomy were collected and analyzed. RESULTS The mean age and BMI of the patients were 33.1 years and 26.1 kg/m2, respectively. 64% of patients did not have any pregnancy and 33% had a history of one or more abortions. The most frequent indications for myomectomy were infertility (40%) and abnormal uterine bleeding (53%). The most frequent location of the fibroids was the intramural fundal (46%), with an average size of 6.5 cm (SD ± 3.6). The number of myomas extracted ranged from 1 to 6. The mean operative time was 107.8 min (SD ± 27.5) and the operative bleeding was 145.4 mL (SD ± 42.2). There were no operative complications. The conversion rate to laparotomy was 2%. Power morcellation was used in all cases without complications. CONCLUSIONS Laparoscopic myomectomy with electromechanical morcellator for tissue extraction in low risk patients for malignancy is a feasible procedure with a low conversion rate, minimal operative bleeding and the operative time similar to that reported for myomectomy performed by laparotomy.


Towards a better world after COVID-19.


COVID-19: the new challenge for rheumatologists.


title: Insights in Public Health: COVID-19 Special Column: The Crisis of Non-Communicable Diseases in the Pacific and the Coronavirus Disease 2019 Pandemic.

text: Globally, coronavirus disease 2019 (COVID-19) is threatening human health and changing the way people live. With the increasing evidence showing comorbidities of COVID-19 and non-communicable diseases (NCDs), the Pacific region, where approximately 75% of deaths are due to NCDs, is significantly vulnerable during this crisis unless urgent action is taken. Whilst enforcing the critical mitigation measures of the COVID-19 pandemic in the Pacific, it is also paramount to incorporate and strengthen NCD prevention and control measures to safeguard people with NCDs and the general population; keep people healthy and minimise the impact of COVID-19. To sustain wellbeing of health, social relationships, and the economy in the Pacific, it is a critical time for all governments, development partners and civil societies to show regional solidarity in the fight against emerging COVID-19 health crisis and existing Pacific NCDs crisis through a whole of government and whole of society approach.


title: Bovine coronavirus structural proteins.

text: The tissue culture-adapted strain (Mebus) of bovine coronavirus was grown in the presence of isotopically labeled amino acids, glucosamine, or orthophosphate for the purpose of analyzing the virion structural proteins. Five species of polypeptides were identified when purified virions were solubilized in urea and sodium dodecyl sulfate and resolved by polyacrylamide gel electrophoresis. Four species were glycosylated and had apparent molecular weights of 140,000, 120,000, 100,000, and 26,000. The glycoproteins were susceptible to proteolytic cleavage and enzymatic iodination when intact virions were studied and are thus at least partially external to the virion envelope. The 140,000-molecular-weight glycoprotein is apparently a dimer of 65,000-molecular-weight glycopolypeptides held together by disulfide linkages. Species 5 was phosphorylated and had an apparent molecular weight of 52,000. In the intact virion, it was unaffected by protease and was not enzymatically iodinated. It is therefore apparently an internal protein.


title: Coronavirus leader RNA regulates and initiates subgenomic mRNA transcription both in trans and in cis.

text: Mouse hepatitis virus (MHV), a coronavirus, utilizes a discontinuous transcription mechanism for subgenomic mRNA synthesis. Previous studies (C.-L. Liao and M. C. C. Lai, J. Virol. 68:4727-4737, 1994) have demonstrated that an upstream cis-acting leader sequence serves as a transcriptional enhancer, but the mechanism of transcriptional regulation is not clear. In this study, we constructed a series of defective interfering (DI) RNAs containing the chloramphenicol acetyltransferase (CAT) gene behind a differentially expressed transcription initiation (intergenic) sequence (for mRNA2-1). These DI RNAs had different copy numbers of the UCUAA pentanucleotide sequence at the 3' end of the leader. Transfection of these DI RNA constructs into cells infected with a helper MHV, which contains either two or three UCUAA copies at the 3' end of the leader, resulted in differential expression of CAT activities. We demonstrated that the copy number of UCUAA repeats in the leaders of both helper viral and DI RNAs affected the level of CAT activity, suggesting that MHV leader RNA could regulate both in trans and in cis the transcription of subgenomic mRNAs. The leader RNA of subgenomic mRNAs was derived from either the trans- or the cis-acting leader. Furthermore, insertion of a UA-rich sequence (UUUAUAAAC) immediately downstream of the leader in DI RNA, to match the sequence of helper viral RNA, enhanced the CAT activity by threefold, suggesting that this nine-nucleotide sequence is a cis-acting element. Interestingly, when the nine-nucleotide sequence was absent in DI RNA, the leaders of subgenomic mRNAs were exclusively derived from the helper virus. In contrast, when the nine-nucleotide sequence was present in DI RNA, the leaders were derived from both helper viral and DI RNAs. These results suggest that the nine-nucleotide sequence either is required for the leader RNA to initiate mRNA synthesis or, alternatively, serves as a transcription terminator for the leader RNA synthesis. However, when a constitutively expressed intergenic sequence (for mRNA7) was used, no difference in transcription efficiency was noted, regardless of the copy number of UCUAA in the DI RNA and helper virus. This study thus indicates that MHV subgenomic RNA transcription requires the interaction among the intergenic (promoter) sequence, a trans-acting leader, and a cis-acting leader sequence. A novel model of transcriptional regulation of coronavirus subgenomic mRNAs is presented.


title: [Antigenicity analysis of nucleocapsid proteins of 3 human coronaviruses SARS-CoV, 229E and OC43 with their monoclonal antibodies].

text: OBJECTIVE To prepare and characterize monoclonal antibodies (mAbs) against the recombinant nucleocapsid (N) protein of 3 human coronaviruses SARS-CoV, 229E and OC43 and study the antigenic relationship between the 3 N proteins. METHODS BALB/c mice were immunized with the recombinant N proteins of SARS-CoV, 229E and OC43 to obtain the mAbs by means of hybridoma. Screening and identification of the mAbs were performed using indirect enzyme-linked immunosorbent assay (ELISA), Western blotting and indirect immunofluorescence assay. Cross-reactivity between the N proteins of the 3 coronaviruses was analyzed with the prepared mAbs. RESULTS The mAbs against the recombinant N proteins of SARS-CoV, 229E and OC43 were obtained, which reacted specifically with the corresponding viral N protein as shown by indirect ELISA, Western blotting and indirect immunofluorescence assay. No cross-reactivity was found between the 3 N proteins. CONCLUSION The prepared mAbs against the recombinant N proteins may provide valuable assistance in studying antigenic relationships of N proteins between the 3 human coronaviruses.


Antigen processing.


title: Toxoplasma gondii: from animals to humans.

text: Toxoplasmosis is one of the more common parasitic zoonoses world-wide. Its causative agent, Toxoplasma gondii, is a facultatively heteroxenous, polyxenous protozoon that has developed several potential routes of transmission within and between different host species. If first contracted during pregnancy, T. gondii may be transmitted vertically by tachyzoites that are passed to the foetus via the placenta. Horizontal transmission of T. gondii may involve three life-cycle stages, i.e. ingesting infectious oocysts from the environment or ingesting tissue cysts or tachyzoites which are contained in meat or primary offal (viscera) of many different animals. Transmission may also occur via tachyzoites contained in blood products, tissue transplants, or unpasteurised milk. However, it is not known which of these routes is more important epidemiologically. In the past, the consumption of raw or undercooked meat, in particular of pigs and sheep, has been regarded as a major route of transmission to humans. However, recent studies showed that the prevalence of T. gondii in meat-producing animals decreased considerably over the past 20 years in areas with intensive farm management. For example, in several countries of the European Union prevalences of T. gondii in fattening pigs are now <1%. Considering these data it is unlikely that pork is still a major source of infection for humans in these countries. However, it is likely that the major routes of transmission are different in human populations with differences in culture and eating habits. In the Americas, recent outbreaks of acute toxoplasmosis in humans have been associated with oocyst contamination of the environment. Therefore, future epidemiological studies on T. gondii infections should consider the role of oocysts as potential sources of infection for humans, and methods to monitor these are currently being developed. This review presents recent epidemiological data on T. gondii, hypotheses on the major routes of transmission to humans in different populations, and preventive measures that may reduce the risk of contracting a primary infection during pregnancy.


title: Carbohydrate microarrays.

text: Sugar chains are abundantly expressed on the outer surfaces of the vast majority of viral, bacterial, protozoan and fungal pathogens, as well as on the membranes of mammalian cells. This class of carbohydrate molecule is without peer in structural diversity and is characteristically suitable for storing and displaying biological signals for molecular and cellular recognition. Exploring the biological information contained in sugar chains is an important topic of current postgenomic research. To facilitate these investigations, we have focused on the establishment of a carbohydrate-based microarray technology. Recently, we reported that a large panel of carbohydrate-containing macromolecules, including polysaccharides, natural glycoconjugates, and the mono- and oligosaccharides coupled to carrier molecules, can be stably immobilized on a microglass slide to produce a large-scale carbohydrate microarray. In this review, we attempt to summarize our recent progress in using this technology to uncover the carbohydrate-based biological signals that are recognized by the human and animal immune systems. We also discuss the potential of various platforms of carbohydrate microarrays that were recently established and analyze the challenges to future development of carbohydrate microarray technologies and their applications.


title: Fowlpox virus as a recombinant vaccine vector for use in mammals and poultry.

text: Live vaccines against fowlpox virus, which causes moderate pathology in poultry and is the type species of the Avipoxvirus genus, were developed in the 1920s. Development of recombinant fowlpox virus vector vaccines began in the 1980s, for use not only in poultry, but also in mammals including humans. In common with other avipoxviruses, such as canarypox virus, fowlpox virus enters mammalian cells and expresses proteins, but replicates abortively. The use of fowlpox virus as a safe vehicle for expression of foreign antigens and host immunomodulators, is being evaluated in numerous clinical trials of vaccines against cancer, malaria, tuberculosis and AIDS, notably in heterologous prime-boost regimens. In this article, technical approaches to, and issues surrounding, the use of fowlpox virus as a recombinant vaccine vector in poultry and mammals are reviewed.


title: [Spontaneous hepatitis A with a fatal outcome in rhesus monkeys].

text: This is the first report of virologically verified spontaneous hepatitis A in M. rhesus monkeys with severe involvement of the liver leading to the death of the animals. In 21 out of 23 dead monkeys morphological lesions in the liver have been characterized as acute hepatitis. In 6 (26%) animals no other pathological processes were found. In 15 animals hepatitis was combined with other diseases (dysentery, parasitic infestations, coronavirus infection). Antigen of hepatitis A virus was detected by an enzyme immunoassay in the intestinal contents of 8 monkeys and in the livers of 3 of them. Immune electron microscopic studies detected in the intestines some virus particles morphologically and antigenically similar to human hepatitis A virus.


title: Expression of the porcine transmissible gastroenteritis coronavirus M protein.

text: Cloned cDNA encoding the M protein of the porcine transmissible gastroenteritis coronavirus (TGEV) was introduced into a vaccinia virus to examine the function of the amino-terminal signal peptide. The M protein expressed by the recombinant virus was targeted to the Golgi region of infected cells, as is the M protein in cells infected with TGEV. The protein appeared not to undergo processing other than glycosylation. However, the vaccinia-expressed M protein was slightly larger than the protein found in TGEV-infected cells, suggesting that a difference in modification exists between the proteins.


title: [Expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E.coli and identification of its mimic antigenicity].

text: AIM To study the expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E.coli and its mimic antigenicity to S2 protein. METHODS B cell epitopes in S2 subunit of SARS coronavirus spike protein was predicted using DNAStar software. The cDNA sequence encoding the B cell epitope peptide was constructed artificially by PCR and then cloned into the downstream of chaperone 10 gene in vector pET28a(+) to construct pET28-chap10-S2epi plasmid. The fusion protein, chap10-S2epi, was expressed in E.coli BL21(DE3) and identified by SDS-PAGE and Western blot. The rabbit was immunized by purified Chap10-S2epi for the preparation of antiserum, which was used to identify the mimic antigenicity of Chap10-S2epi to S2 protein by ELISA. RESULTS Chap10-S2epi fusion protein was successfully constructed and expressed in E.coli. The antiserum from the animal immunized by Chap10-S2epi recognized full length of SARS coronavirus S2 spike protein. CONCLUSION The predicted B cell epitope peptide of SARS coronavirus S2 spike protein can induce the antigenicity of S2 protein, which provides some fundamental data for developing engineering vaccine against SARS coronavirus infection.


title: Physical medicine and rehabilitation conditions in the Astrodome clinic after hurricane Katrina.

text: OBJECTIVE To report the physical medicine and rehabilitation (PMR) conditions seen in the Astrodome Clinic after Hurricane Katrina. DESIGN Retrospective chart analysis from the county hospital-sponsored disaster-relief clinic in large urban city, including a study of 239 patients with 292 PMR conditions. The total number of patients seen in the Astrodome Medical Clinic was 11,245. The Astrodome database was reviewed for PMR condition diagnostic codes. A retrospective chart analysis was conducted, including date of visit, age, gender, ethnicity, and PMR diagnosis category. Descriptive statistics were obtained for the entire sample. chi2 or t tests were used to determine gender, age, or date-of-service predominance for the most common diagnostic categories. RESULTS Mean +/- SD age was 45.7 +/- 14.3 yrs; 56% were women, 43% were men (1% unspecified), and 76% were African American. The majority (75%) of PMR conditions presented in the first week. Most frequent were swollen feet and legs (22%), leg pain and cramps (17%), headache (12%), and neck and back pain (10%). Persons with headaches were younger than those without (41.3 vs. 46.3 yrs, P = 0.048). Persons with neck and/or back pain were older than those without those conditions (51.3 vs. 44.8 yrs, P = 0.004). Women had more headaches (20.9%) than did men (6.7%, P = 0.002). There were no Caucasians with leg pain/cramps, whereas 20.2% of African Americans had this condition (P = 0.028). CONCLUSIONS This study documents the time of clinic presentation and most frequent types of PMR conditions of patients treated in the Astrodome Clinic after a historic hurricane. Most PMR conditions were treated by PMR personnel during the first week. Thus, future disaster planning should include PMR professionals as early responders.


Medical humanities and medical informatics: an unlikely alliance? Is there a role for patients' voices in the modern case record?


title: Reovirus-like agent associated with neonatal diarrhea in pronghorn antelope.

text: Reovirus-like particles were demonstrated by negative stain electron microscopic examination of the feces from antelope fawns with diarrhea. Fluorescent antibody tests on frozen sections of ileum from one dead antelope fawn and immunoelectron microscopy tests on feces from two live fawns provided evidence that the antelope agent was serologically related to the neonatal calf diarrhea reovirus-like agent.


title: Altered p38 mitogen-activated protein kinase expression in different leukocytes with increment of immunosuppressive mediators in patients with severe acute respiratory syndrome.

text: Severe acute respiratory syndrome (SARS) has spread to a global pandemic, especially in Asia. The transmission route of SARS has been clarified, but the immunopathogenesis of SARS is unclear. In an age-matched case-control design, we studied immune parameters in 15 SARS patients who were previously healthy. Plasma was harvested for detection of virus load, cytokines, and nitrite/nitrate levels, and blood leukocytes were subjected to flow cytometric analysis of intracellular mitogen-activated protein kinases (MAPKs) in different leukocytes. Patients with SARS had significantly higher IL-8 levels (p = 0.016) in early stage, and higher IL-2 levels (p = 0.039) in late stage than normal controls. Blood TNF-alpha, IL-6, and IL-10, and nitrite/nitrate levels were not significantly elevated. In contrast, TGF-beta and PGE(2) levels were significantly elevated in SARS patients. Five of the 15 SARS patients had detectable coronaviruses in blood, but patients with detectable and undetectable viremia had no different profiles of immune mediators. Flow cytometric analysis of MAPKs activation by phospho-p38 and phospho-p44/42 (extracellular signal-regulated kinase) expression showed that augmented p38 activation (p = 0.044) of CD14 monocytes associated with suppressed p38 activation (p = 0.033) of CD8 lymphocytes was found in SARS patients. These results suggest that regulation of TGF-beta and PGE(2) production and MAPKs activation in different leukocytes may be considered while developing therapeutics for the SARS treatment.


title: Turning over a new leaf.

text: ADDENBROOKE'S trust in Cambridge has launched its own intranet Asite. Set up in January 1999, thew internal network is known as Bluepages and is part of the trust's Information strategy. To access Bluepages, all wards and clinical departments throughout the trust have at least two computers - manyhave three or more.


title: Emergency preparedness for biological and chemical incidents: a survey of anesthesiology residency programs in the United States.

text: UNLABELLED We surveyed health care professionals about their preparations to manage the clinical problems associated with patients exposed to hazardous substances, including weapons of mass destruction (WMD). Training for WMD is considered a key part of public health policy and preparedness. Although such events are rare, when they do occur, they can cause mass casualties. In many models of mass casualty management, anesthesiology personnel are responsible for treating patients immediately on arrival at the hospital. We studied the extent of training offered to anesthesiology personnel in the use of WMD protective gear and patient management in United States (US) anesthesiology residency programs. Information was obtained via an online survey to all program directors and chair persons of anesthesiology programs. We polled all of the 135 US anesthesiology programs of which 90 (67%) responded. Only 37% had any form of training, and many of them did not repeat training after initial sessions. Twenty-eight percent of programs east of the Mississippi River reported some form of training whereas only 17% of programs west of it reported training available. The majority of anesthesia residency programs in the US that responded to our survey provided little or no training in the management of patients exposed to WMD. IMPLICATIONS In an attack involving weapons of mass destruction or toxic chemicals, anesthesiologists will provide care. Our survey of United States anesthesiology residency programs demonstrated that there is limited training of residents regarding the anesthetic management of patients injured by weapons of mass destruction. This has serious public health implications.


WHO is worried that China is under-reporting SARS.


title: Vanishing vaccinations: why are so many Americans opting out of vaccinating their children?

text: Vaccinations against life-threatening diseases are one of the greatest public health achievements in history. Literally millions of premature deaths have been prevented, and countless more children have been saved from disfiguring illness. While vaccinations carry unavoidable risks, the medical, social and economic benefits they confer have led all fifty states to enact compulsory childhood vaccination laws to stop the spread of preventable diseases. Today, however, vaccines are becoming a victim of their success--many individuals have never witnessed the debilitating diseases that vaccines protect against, allowing complacency toward immunization requirements to build. Antivaccination sentiment is growing fast in the United States, in large part due to the controversial and hotly disputed link between immunizations and autism. The internet worsens fears regarding vaccination safety, as at least a dozen websites publish alarming information about the risks of vaccines. Increasing numbers of parents are refusing immunizations for their children and seeking legally sanctioned exemptions instead, apparently fearing vaccines more than the underlying diseases that they protect against. A variety of factors are at play: religious and philosophical beliefs, freedom and individualism, misinformation about risk, and overperception of risk. State legislatures and health departments now face a difficult challenge: respecting individual rights and freedoms while also safeguarding the public welfare. Nearly all states allow vaccination exemptions for religious reasons and a growing number provide "philosophical" opt-outs as well. However, in all but a handful of jurisdictions, neither objection is seriously documented or verified. Often, the law requires a parent to do no more than simply check a box indicating she does not wish her child to receive immunizations. The problem is exacerbated by financial incentives schools have to encourage students to opt out of vaccinations. The rise in parents opting out has caused the AMA grave concern, with many experts decrying the rise of so-called "exemptions of convenience." In some areas, nearly one out of five children have not received their recommended vaccines. The consequences are serious not only for those unprotected children, but for the rest of society as well. "Herd immunity" is threatened as more and more parents free ride off of the community's dwindling immunity, and outbreaks of diseases thought to have been conquered have already occurred. Lawsuits against vaccine manufacturers threaten them with bankruptcy, costs are being externalized onto the healthcare and legal systems, and vulnerable populations are suffering harm or even death. In the interests of social welfare, state legislatures and health departments should consider methods to ensure that the exemption process is carefully tailored to prevent check-the-box opt-outs of convenience, while still allowing exemptions for those with earnest and informed convictions or medical reasons.


[Classification and nomenclature of viruses].


title: Disease surveillance among newly arriving refugees and immigrants--Electronic Disease Notification System, United States, 2009.

text: PROBLEM/CONDITION Approximately 450,000 legal permanent immigrants and 75,000 refugees enter the United States annually after receiving required medical examinations by overseas panel physicians (physicians who follow the CDC medical screening guidelines provided to the U.S. Department of State). CDC has the regulatory responsibility for preventing the introduction, transmission, and spread of communicable diseases into the United States as well as for developing the guidelines, known as technical instructions, for the overseas medical examinations. Other conditions that are not infectious might preclude an immigrant or refugee from entering the United States and also are reported as part of the medical examination. After arrival in the United States, all refugees are recommended to obtain a medical assessment by a health-care provider or a health department within 30 days. In addition, immigrants with certain medical conditions such as noninfectious tuberculosis at the time of the original medical examination are recommended to be evaluated after arrival to ensure that appropriate prevention or treatment measures are instituted. Health departments need timely and accurate notifications of newly arriving immigrants, refugees, and persons with other visa types to facilitate these evaluations. Notifications for all newly arriving refugees (with or without medical conditions) and immigrants with medical conditions are provided by CDC's Electronic Disease Notification (EDN) system. This is the first report describing EDN. REPORTING PERIOD This report summarizes notifications by the EDN system during January-December 2009. DESCRIPTION OF SYSTEM The EDN system is a centralized electronic reporting system that collects health information on newly arriving refugees and immigrants with Class A and Class B medical conditions. Class A conditions render applicants inadmissible and require a waiver for entry; Class B conditions are admissible but might require treatment or follow-up. Information in the EDN system is used to notify state health departments in all 50 states and the District of Columbia about the arrival of these persons in the United States. RESULTS In 2009, the EDN system notified U.S. state and local health departments of 104,954 newly arriving refugees and immigrants, of whom 78,899 (75.2%) were refugees (with or without medical conditions), 19,358 (18.4%) were immigrants with medical conditions, and 6,697 (6.4%) were persons with other visa types. Of the 78,899 refugees, 21,319 (27%) had a medical condition. The majority (93.4%) of immigrants with medical conditions had tuberculosis classifications (i.e., either had evidence of latent tuberculosis infection or chest radiograph findings interpreted by the overseas panel physician as consistent with tuberculosis). Of the 41,415 refugees and immigrants with Class A or Class B medical conditions, 405 (1%) had Class A conditions, and 40,994 (99%) had Class B conditions. The majority of refugees and immigrants with suspected Class B tuberculosis were born in the Philippines (41.3%), Mexico (12.1%), Burma (8.7%), Vietnam (7.8%), and the Dominican Republic (5.8%). The majority of refugee notifications were for persons born in Iraq (23.9%), Burma (18.9%), and Bhutan (15.1%). Approximately one third of the tuberculosis notifications were sent to health departments in California (20.5%), Texas (9.8%), and New York (6.3%), and the national reporting rate for tuberculosis follow-up was 75.4% within 30 days of arrival. INTERPRETATION The findings in this report suggest that 1) overseas medical screening results in a low frequency (0.4%) of inadmissible medical conditions in the United States, 2) the EDN system provides more direct notifications to health departments than the previous paper-based system about newly arriving immigrants and refugees who need medical follow-up, and 3) approximately 75% of follow-up occurs among persons with suspected tuberculosis who are reported to EDN by states receiving newly arriving refugees and immigrants. PUBLIC HEALTH ACTIONS The data in this report can be used to help state and local health departments provide prompt and effective follow-up, evaluation, and treatment to newly arriving immigrants and refugees. Timely follow-up might prevent additional spread of tuberculosis or other communicable diseases of public health significance into their communities. In addition, information from the EDN system allows health departments to use their resources as effectively as possible by providing clinical information that identifies the refugees and immigrants who should be prioritized for evaluation and treatment.


title: Cloning and quantification of ferret serum amyloid A.

text: Serum amyloid A (SAA) is used as a biomarker for infections and inflammation in humans and veterinary medicine. We cloned ferret cDNA encoding SAA from the liver of a ferret via reverse transcription PCR (RT-PCR). The sequence of the cDNA clone revealed that ferret SAA has an open reading frame of 387 bp that encodes 129 amino acids. The deduced amino acid sequence of ferret SAA has 96.1, 89.9, 86.0, 83.8, 83.0, 73.8 and 65.3% similarity to the mink, dog, cat, cattle, horse, human and mouse SAA genes, respectively. Compared to human SAA, the deduced ferret SAA amino acid sequence had an insertion of an 8-amino acid fragment between amino acids 88 and 95. Recombinant ferret SAA (rfrSAA) was expressed using an Escherichia coli (E. coli) strain, BL21 Star. Using Western blot analysis, anti-SAA mAb provided with the multispecies SAA ELISA kit reacted with purified rfrSAA. A significant dose-response relationship was observed between the rfrSAA protein and a commercial multispecies SAA ELISA kit. In contrast, rfrSAA was not recognized with the antibodies included in a commercial human SAA ELISA kit. These results suggest that the structure of ferret SAA is antigenically similar to other domestic animal SAAs, and the multispecies ELISA kit allows for the detection and quantification of ferret SAA in vivo.


title: Idiopathic granulomatous mastitis in Hispanic women - Indiana, 2006-2008.

text: Idiopathic granulomatous mastitis (IGM) is a rare inflammatory breast lesion of unknown etiology that occurs in women of childbearing age; only a few hundred cases have been reported worldwide. A breast cancer mimic, IGM also is diagnosed by breast biopsy. On December 12, 2008, a physician in Indianapolis, Indiana, reported a cluster of seven IGM diagnoses in multigravid Hispanic women, an unusually high number. To evaluate the etiology of the breast masses and characterize associated epidemiologic and clinical features, the Indiana State Department of Health and CDC conducted a multidisciplinary investigation. This report describes the results of that investigation. A total of nine cases of IGM were confirmed during 2006-2008 in Indianapolis, for an annual prevalence of IGM of 2.4 per 100,000 women aged 20-40 years. The prevalence was 12 times higher among Hispanic women. Among IGM patients at the hospital, a median of 5 months elapsed between symptom onset and diagnostic biopsy. Histopathologic evaluations confirmed IGM. In a case-control study of all seven cases and 21 controls from the hospital, case-patients were significantly more likely than controls to have less than a sixth-grade education (odds ratio [OR] = 12.7), a positive tuberculin skin test (OR = undefined), or a medication allergy (OR = 15.0). No other risk factors were significantly associated with case status. Barriers to accessing health care, including low education level, resulted in delayed care for breast masses. Future research could provide more complete descriptions of the epidemiology and etiology of IGM.


Public Reporting of COVID-19 Management.


The future of antiviral chemotherapy.


title: Vacuolar degeneration in mice infected with a coronavirus JHM-CC strain.

text: Vacuolar degeneration was constantly induced in the CNS of 4-week-old ICR mice by intracerebral or intranasal inoculation of JHM-CC virus, a small plaque mutant of mouse hepatitis virus (JHM). Most animals showed no symptoms or only mild hindlimb paresis. Irrespective of clinical manifestations, the virus was isolated from the CNS up to days 14 to 16. Viral antigen expression in the CNS tissue was most extensive around days 5 to 7 and became undetectable on day 14. Viral antigens were localized almost exclusively to neurons, and the temporal sequence of viral antigen distribution after intranasal inoculation clearly indicated the virus spread through the olfactory and limbic systems into the brainstem and spinal cord, and possible cell-to cell transmission of the virus within the CNS. Vacuolar changes, most conspicuous in the brainstem and spinal cord, were steadily progressive up to 4 weeks after infection, but became indistinct by 4 months. Although the distribution of vacuolar lesions largely agreed with that of viral antigen-positive cells, the severity of vacuolation did not correlate with that of inflammation. Intramyelinic splitting, periaxonal edema, and swollen neurites were major ultrastructural substrates for vacuolar changes. This model could provide a better understanding of new types of neurologic disorders associated with viral infections, including vacuolar myelopathy in AIDS.


title: Avian flu: the creation of expectations in the interplay between science and the media.

text: This paper examines the emerging cultural patterns and interpretative repertoires in reports of an impending pandemic of avian flu in the UK mass media and scientific journals at the beginning of 2005, paying particular attention to metaphors, pragmatic markers ('risk signals'), symbolic dates and scare statistics used by scientists and the media to create expectations and elicit actions. This study complements other work on the metaphorical framing of infectious disease, such as foot and mouth disease and SARS, tries to link it to developments in the sociology of expectations and applies insights from pragmatics both to the sociology of metaphor and the sociology of expectations.


title: The American Society for Virology--29th Annual Meeting.

text: The American Society for Virology 29th Annual Meeting, held in Bozeman, MT, USA, included topics covering new vaccine technologies, delivery methods and treatments in the field of virology. This conference report highlights selected presentations on human norovirus (HuNoV), SARS coronavirus and Rift Valley fever virus vaccine technologies; programmed cell death-1 (PD1) blockade and HyperAcute alpha-Gal platform technology methods; aerosol vaccination delivery; novel technologies to produce influenza virus-like particles (VLP) in mammalian cell lines; and investigational human rotavirus vaccines.


title: The introduction of a laparoscopic liver surgery programme: a cost analysis of initial experience in a university hospital.

text: BACKGROUND AND AIMS In the era of expanding costs of healthcare, this study was conducted to perform a cost analysis of introducing a laparoscopic liver surgery programme for left sided liver lesions. MATERIALS AND METHODS Consecutive patients treated by laparoscopic liver resections of left lateral segments were included. Controls were a group of 14 patients undergoing open resection for similar pathology. Primary outcomes were costs. Secondary outcomes were complications, conversions, blood loss, length of operation, and length of hospital stay. RESULTS The laparoscopic approach for hepatic left lateral resection (bisegmentectomy 2 and 3) was performed in fourteen patients (group I, median age 54 [range 26-82] years). In the open group, fourteen patients from a prospectively collected database with the same type of resection were selected (group II, median age 64 [range 29-76] years). Costs of theatre usage in the laparoscopic group were significantly lower (p=0.031). No significant differences in costs of disposable instruments, ward stay and total costs were observed between the two groups. There were three complications in the laparoscopic group compared with two complications in the open group. In the laparoscopic group there were 2 conversions (14%). Median blood loss was significantly lower in the laparoscopic group (50 mls [range 0-750], (p=0.001) versus the open group (500 mls [range 150-750]). Furthermore, operation time was also significantly lower in the laparoscopic group (116 [range 85-261] minutes) versus the open group (165 [range 96-217] minutes, p=0.016). Median length of stay was 6 [range 4-11] days in group I versus 6 [range 5-13] days in group II (p=0.508). CONCLUSION Costs of laparoscopic liver resections proved to be equivalent to open surgery. Furthermore, implementation of a laparoscopic liver resection programme seems feasible and safe with reduced blood loss and operation time and comparable morbidity and length of stay.


title: [Severe acute respiratory syndrome. The public health laboratory in a global emergency].

text: By the end of year 2002 there was an outbreak of atypical pneumonia in Southeast Asia which soon spread to other continents. This new severe acute respiratory syndrome (SARS) was produced by a novel coronavirus. Due to the severity of the situation and risk of introduction of this pathology in our country, the need to arrange specific laboratory diagnostic tests arose. Classic techniques, such as the electron microscopy and molecular biology test such as retrotranscription followed by the polymerase chain reaction (RT-PCR) were implemented. The araldit included cells infected with bovine coronavirus which allowed the viral particles to be visualized easily but it took more time in comparison with the negative staining of free particles from viral cultures. RT-PCR was able to detect RNA of isolated viruses from cases in Hong Kong and Germany.


title: Heterogeneity of systematic reviews in oncology.

text: Systematic reviews synthesize data across multiple studies to answer a research question, and an important component of the review process is to evaluate the heterogeneity of primary studies considered for inclusion. Little is known, however, about the ways that systematic reviewers evaluate heterogeneity, especially in clinical specialties like oncology. We examined a sample of systematic reviews from this body of literature to determine how meta-analysts assessed and reported heterogeneity. A PubMed search of 6 oncology journals was conducted to locate systematic reviews and meta-analyses. Two coders then independently evaluated the manuscripts for 10 different elements based on an abstraction manual. The initial PubMed search yielded 337 systematic reviews from 6 journals. Screening for exclusion criteria (nonsystematic reviews, genetic studies, individual patient data, etc.) found 155 articles that did not meet the definition of a systematic review. This left a final sample of 182 systematic reviews across 4 journals. Of these reviews, 50% (91/182) used varying combinations of heterogeneity tests, and of those, 16% (15/91) of review authors noted excessive heterogeneity and opted to not perform a meta-analysis. Of the studies that measured heterogeneity, 51% (46/91) used a random-effects model, 7% (8/91) used a fixed-effects model, and 43% (39/91) used both. We conclude that use of quantitative and qualitative heterogeneity measurement tools are underused in the 4 oncology journals evaluated. Such assessments should be routinely applied in meta-analyses.


title: EDs on heightened alert for MERS-CoV as first cases reach the US.

text: The first cases of Middle East Respiratory Syndrome coronavirus (MERS-CoV) have turned up in the United States. First, in late April, a patient tested positive for the virus at a hospital in Munster, IN, and then shortly thereafter, a second patient tested positive at a hospital in Orlando, FL. While both patients have since recovered from the virus and been released, the cases have raised awareness of the infectious threat of MERS-CoV, and they have put EDs and other frontline providers on heightened alert for patients with severe respiratory symptoms and other risk factors. While MERS-CoV is not yet as contagious as seasonal influenza or the severe acute respiratory syndrome (SARS) that started in China and then swept around the globe in 2003, it is more deadly. The World Health Organization reports that roughly one-quarter of 514 people who have tested positive for the virus have died. Experts note that health care workers make up a large percentage of the documented cases of MERS-CoV, and they point out that most human-to-human transmissions of the virus occur in the hospital setting. Public health officials urge emergency personnel to pay strict attention to infection control practices, and to query patients who present with fever and respiratory distress about recent travel to the Arabian Peninsula and/or close contact with a person who has a confirmed or probable case of MERS-CoV.


title: Detection of porcine reproductive and respiratory syndrome virus in cell cultures and formalin-fixed tissues by in situ hybridization using a digoxigenin-labeled probe.

text: A nonradioactive in situ hybridization method is described for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) in cell cultures and in formalin-fixed paraffin-embedded tissue sections originating from experimentally infected pigs and from 1 field case. A 174 bp cDNA probe targeting the viral RNA encoding the nucleocapsid protein of a Canadian PRRSV isolate was generated by polymerase chain reaction. The cDNA probe was labeled by random priming with digoxigenin-dUTP using a commercially available kit. The ability of the digoxigenin-labeled probe to specifically detect PRRSV RNA was tested on cultured cells infected with 6 Canadian PRRSV isolates, a US PRRSV isolate and the European Lelystad isolate. The probe detected all Canadian PRRSV isolates tested as well as the US PRRSV isolate but did not detect the Lelystad isolate. In addition, when tested on formalin-fixed paraffin-embedded tissue sections from pigs experimentally infected with several Canadian isolates and from a field case, a strong signal without background staining was obtained. Our results indicate that nonradioactive in situ hybridization could represent a useful tool for the detection of PRRSV in routinely fixed and processed tissues. In situ hybridization could also be used to differentiate infection by North American and European Lelystad-like PRRSV isolates.


Managing the community response to bioterrorist threats.


title: Improving access to outpatient cardiac care at the national heart centre--a partnership between specialists and primary care.

text: Ensuring timely access to specialist care is an important indicator of the quality of a health service. Demand for cardiology outpatient appointments has grown considerably in the last decade, leading to increased waiting time for cardiology appointments at public hospitals. This paper examines the effectiveness of past and ongoing strategies initiated by the National Heart Centre, many of which were in collaboration with SingHealth Polyclinics, documents the lessons learnt, and provides a framework for approaching this problem. Instead of a simplistic approach where institutions react to long waiting times by growing capacity to meet demand, this paper emphasises the need to focus on the final intended outcome (timely diagnosis and treatment) rather than on a single performance indicator, such as waiting time. A broad systems approach at the national level is advocated, rather than piecemeal, uncoordinated actions by individual hospitals.


title: [LAPAROSCOPIC PARTIAL PERICYSTECTOMY OF SPLENIC HYDATID CYST - A CASE REPORT].

text: Echinococcal cyst of the spleen is usually a result of infection with the parasite Echinococcus granulosus. The spleen is the third most frequent localization of echinococcus after liver and lungs. Partial laparoscopic pericystectomy can be done without the loss of blood and scattering of scolexes with spleen preservation and conservation of its immune function. We present the patient with a large (1 8 x 16 x 12 cm) echinococcal cyst of the spleen that compressed the surrounding organs (stomach, transverse colon, pancreas and left kidney), and prevented normal passage causing vomiting after every meal. In this patient, PAIR procedure (puncture, aspiration, injection, reaspiration) and conservative treatment was attempted on several occasions without success. Finally, laparoscopic partial pericystectomy was performed, evacuating the contents of the cyst. The surgery lasted 120 minutes. Postoperatively the patient was without complications. Hospitalization lasted five days. Six months later, the patient is without problems. These echinococcus cysts of the spleen cannot be solved using PAIR technique and conservative treatment. Laparoscopic partial pericystectomy is a better solution than open surgery due to less trauma to the patient, especially in elderly people.


title: Fatal cases of Rocky Mountain spotted fever in family clusters--three states, 2003.

text: Rocky Mountain spotted fever (RMSF), a tickborne infection caused by Rickettsia rickettsii and characterized by a rash, has a case-fatality rate as high as 30% in certain untreated patients. Even with treatment, hospitalization rates of 72% and case-fatality rates of 4% have been reported. This report summarizes the clinical course of three fatal cases of RMSF in children and related illness in family members during the summer of 2003. These cases underscore the importance of 1) prompt diagnosis and appropriate antimicrobial therapy in patients with RMSF to prevent deaths and 2) consideration of RMSF as a diagnosis in family members and contacts who have febrile illness and share environmental exposures with the patient.


title: Airborne spread of disease--the implications for dentistry.

text: The potential for the airborne spread of disease has been recognized for many years. Recent studies have shown that this mode for disease transmission is capable of spreading a fatal disease such as Severe Acute Respiratory Syndrome over a wide area. Many dental procedures produce extensive aerosols and splatter that are routinely contaminated with bacteria, viruses, and blood. In the past, the potential for these aerosols and splatter to be a vector for disease spread has not been emphasized in dental infection control. Recently published data shows a need to reassess the potential for dental aerosols and splatter to spread disease and the need for their control. Simple and inexpensive methods for the control of dental aerosols and splatter are given. Dental personnel are urged to make the control of aerosols a standard part of their infection control procedures.


title: Contrast enhanced ultrasound and computer tomography diagnosis of solid and mixed pancreatic tumors - analysis of confounders.

text: AIMS. The aim of this study was to evaluate the leading causes of discrepancies between imaging studies [contrast enhanced ultrasound (CEUS) and contrast enhanced computer tomography (CECT)] diagnosis and histology in patients presenting to a tertiary referral center with previously detected pancreatic masses by standard abdominal ultrasound. METHODS. We performed a prospective longitudinal observational study on 76 patients with pancreatic masses: 57 (75%) patients with solid pancreatic tumors and 19 (25%) patients with cystic and mixed pancreatic masses. For each tumor the CEUS and CECT features were analyzed and compared with the final histological diagnosis. RESULTS. Testing the performance of CEUS and CECT in evaluating the benign or malignant etiology of pancreatic masses, we obtained a probability of 82% for CEUS and of 83% for CECT, for a randomly selected individual from the pancreatic tumor group to have an imaging result indicating suspicion for malignancy. We obtained discordances with the histopathological diagnosis in 25 (32.89%) patients for CEUS and in 23 (30.26%) patients for CECT. In multiple regression analysis, two variables independently influenced the discordance between the two imaging methods and histological conclusion: enhancement pattern and tumor nature (solid vs. cystic). CONCLUSIONS.CEUS and CECT showed a good diagnostic performance in differentiating benign from malignant pancreatic tumors. Enhancement pattern and tumor nature (solid vs. cystic) are independent confounders between imaging and histological diagnosis.


title: Lactogenic immunity and milk antibody isotypes to transmissible gastroenteritis virus in sows exposed to porcine respiratory coronavirus during pregnancy.

text: Passive protection provided by sows inoculated with the virulent Miller strain of transmissible gastroenteritis virus (TGEV), or the ISU-1 strain of porcine respiratory coronavirus (PRCV), or both was evaluated in nursing pigs challenge exposed with virulent TGEV. Four sows (group B) were inoculated with PRCV oronasally twice at 4 and 2 weeks before parturition; 1 sow (group C) was inoculated similarly, but in 2 subsequent pregnancies; and 2 sows (group D) were oronasally primed with PRCV at 4 weeks before parturition, and 2 weeks later were administered a booster inoculation of virulent TGEV. Two additional sows (group E) remained uninoculated and served as seronegative controls, and 1 sow (group A) that had been naturally infected with TGEV served as a seropositive control. The degree of passive immunity transferred by these sows to their litters was assessed by challenge exposing the pigs of sows in groups B-E (only the second litter of group C) with virulent TGEV at 3 to 5 days of age. After challenge exposure, clinical signs of infection and mortality were noted and fecal and nasal shedding of virus was assessed by ELISA. The IgA, IgG, and IgM antibody titers to TGEV were quantified in colostrum and milk of the sows by use of an isotype-specific monoclonal antibody-capture ELISA, using biotinylated monoclonal antibodies against each porcine isotype as detecting reagents. A plaque-reduction assay was used to quantify neutralizing antibody titers in serum, colostrum, milk, and fractionated whey (IgG and IgA/IgM). In the sow naturally infected with TGEV (group A), there was a pronounced decrease in IgG antibody titers to TGEV in the transition from colostrum to milk, and IgA TGEV antibodies became predominant, with high titers maintained throughout lactation. The 4 group-B sows partially protected their pigs after TGEV challenge exposure; mean mortality was 67%, compared with 100% in pigs suckling the 2 TGEV seronegative control sows (group-E litters). Although IgA TGEV antibodies were detected in colostrum and milk of group-B sows, IgG TGEV antibodies were the most abundant. The sow of group C had a marked increase in IgA TGEV antibody titers in colostrum and milk after reinoculation with PRCV during the second pregnancy, before TGEV challenge exposure of the litter. Its pigs were passively protected to a high degree after TGEV challenge exposure (27% litter mortality). The sows in group D, primed with PRCV and boosted with TGEV, provided the best passive protection after TGEV challenge exposure of their pigs. Not only litter mortality (27%) but also morbidity was reduced, compared with those factors for the other challenge-exposed litters, and the sows did not become ill.(ABSTRACT TRUNCATED AT 400 WORDS)


title: The psychological impact of the SARS epidemic on hospital employees in China: exposure, risk perception, and altruistic acceptance of risk.

text: OBJECTIVE We examined the psychological impact of the 2003 outbreak of severe acute respiratory syndrome (SARS) on hospital employees in Beijing, China. METHODS In 2006, randomly selected employees (n = 549) of a hospital in Beijing were surveyed concerning their exposure to the 2003 SARS outbreak, and the ways in which the outbreak had affected their mental health. RESULTS About 10% of the respondents had experienced high levels of posttraumatic stress (PTS) symptoms since the SARS outbreak. Respondents who had been quarantined, or worked in high-risk locations such as SARS wards, or had friends or close relatives who contracted SARS, were 2 to 3 times more likely to have high PTS symptom levels, than those without these exposures. Respondents' perceptions of SARS-related risks were significantly positively associated with PTS symptom levels and partially mediated the effects of exposure. Altruistic acceptance of work-related risks was negatively related to PTS levels. CONCLUSIONS The psychological impact of stressful events related to an infectious disease outbreak may be mediated by peoples' perceptions of those events; altruism may help to protect some health care workers against these negative impacts.


title: Impact of Cosmetic Lotions on Nanoparticle Penetration through ex vivo C57BL/6 Hairless Mouse and Human Skin: A Comparison Study.

text: Understanding the interactions of nanoparticles (NPs) with skin is important from a consumer and occupational health and safety perspective, as well as for the design of effective NP-based transdermal therapeutics. Despite intense efforts to elucidate the conditions that permit NP penetration, there remains a lack of translatable results from animal models to human skin. The objectives of this study are to investigate the impact of common skin lotions on NP penetration and to quantify penetration differences of quantum dot (QD) NPs between freshly excised human and mouse skin. QDs were mixed in 7 different vehicles, including 5 commercial skin lotions. These were topically applied to skin using two exposure methods; a petri dish protocol and a Franz diffusion cell protocol. QD presence in the skin was quantified using Confocal Laser Scanning Microscopy. Results show that the commercial vehicles can significantly impact QD penetration in both mouse and human skin. Lotions that contain alpha hydroxyl acids (AHA) facilitated NP penetration. Lower QD signal was observed in skin studied using a Franz cell. Freshly excised human skin was also studied immediately after the sub-cutaneous fat removal process, then after 24 hours rest ex vivo. Resting human skin 24 hours prior to QD exposure significantly reduced epidermal presence. This study exemplifies how application vehicles, skin processing and the exposure protocol can affect QD penetration results and the conclusions that maybe drawn between skin models.


title: Coronavirus disease 2019 in children: Surprising findings in the midst of a global pandemic.

text: Question Coronavirus disease 2019 (COVID-19) is affecting millions of people worldwide. It seems that it affects mostly adults older than 40 years of age, and the death rate is highest for older individuals in the population. What should I tell parents worried about their children contracting the coronavirus (SARS-CoV-2) causing COVID-19, and what symptoms should I look for to determine if there is a need to test for the virus?Answer The COVID-19 global pandemic affects all ages. Severe respiratory manifestations have been the mainstay of illness in adults, with what seems to be rapid deterioration necessitating mechanical ventilation. Only 5% of those tested and found to have COVID-19 have been younger than 19 years, possibly owing to limited testing, as the symptoms in children are usually mild. Symptoms in children include fever, dry cough, rhinorrhea, sore throat, and fatigue, and in 10% diarrhea or vomiting. Rarely dyspnea or hypoxemia were also described. Blood tests and imaging have been shown to be of little value in children and should only be ordered for those in whom you would normally order these investigations for viral-like illness. No specific therapy is available and supportive care with rest, fluids, and antipyretics for children is the recommended approach. Ibuprofen or acetaminophen for fever and pain can be given. Antiviral and immunomodulatory treatment is not recommended at this time for otherwise healthy children, and corticosteroids should also not be used. Children with immunocompromised states should be isolated and avoid contact with others.


title: Cigarette use among high school students - United States, 1991-2009.

text: Understanding the trends in the prevalence of cigarette smoking among youths enables policy makers to target prevention resources more effectively. Every 2 years, CDC analyzes data from the national Youth Risk Behavior Survey (YRBS) to evaluate trends in cigarette use among high school students in the United States. This report updates a previous report and describes results of CDC's 2010 analysis of YRBS data from 1991-2009 for three measures: ever smoked cigarettes, current cigarette use, and current frequent cigarette use. For ever smoked cigarettes, the prevalence did not change from 1991 (70.1%) to 1999 (70.4%), declined to 58.4% in 2003, and then declined more gradually, to 46.3% in 2009. For current cigarette use, the prevalence increased from 27.5% in 1991 to 36.4% in 1997, declined to 21.9% in 2003, and then declined more gradually, to 19.5% in 2009. For current frequent cigarette use, the prevalence increased from 12.7% in 1991 to 16.8% in 1999, declined to 9.7% in 2003, and then declined more gradually, to 7.3% in 2009. For all three measures, rates began to decline in the late 1990s, but the rate of decline slowed during 2003-2009. To increase the rate of decline in cigarette use among high school students, reductions in advertising, promotions, and commercial availability of tobacco products should be combined with full implementation of communitywide, comprehensive tobacco control programs.


title: Evaluation of the surveillance system for undiagnosed serious infectious illness (USII) in intensive care units, England, 2011 to 2013.

text: Emerging infections are a potential risk during mass gathering events due to the congregation of large numbers of international travellers. To mitigate this risk for the London 2012 Olympic and Paralympic Games, a sentinel surveillance system was developed to identify clusters of emerging infections presenting as undiagnosed serious infectious illness (USII) in intensive care units (ICUs). Following a six month pilot period, which had begun in January 2011, the surveillance was operational for a further 18 months spanning the Games. The surveillance system and reported USII cases were reviewed and evaluated after this 18 month operational period including assessment of positive predictive value (PPV), timeliness, acceptability and sensitivity of the system. Surveillance records were used to review reported cases and calculate the PPV and median reporting times of USII surveillance. Sensitivity was assessed through comparison with the pilot period. Participating clinicians completed a five-point Likert scale questionnaire about the acceptability of surveillance. Between 11 July 2011 and 10 January 2013, 34 cases were reported. Of these, 22 remained classified as USII at the time of the evaluation, none of which were still hospitalised. No clusters were identified. The 22 USII cases had no association with the Games, suggesting that they represented the background level of USII in the area covered by the surveillance. This corresponded to an annualised rate of 0.39 cases/100,000 population and a PPV of 65%. Clinicians involved in the surveillance reported high acceptability levels. The USII surveillance model could be a useful public health tool in other countries and during mass gathering events for identifying potential clusters of emerging infections.


title: End-tidal carbon dioxide concentration can estimate the appropriate timing for weaning off from extracorporeal membrane oxygenation for refractory circulatory failure.

text: Although extracorporeal membrane oxygenation (ECMO) is widely used as temporary circulation support, there are no reports of direct parameters indicating cardiac recovery to determine the timing of weaning off. Twenty-five patients supported by ECMO due to hemodynamic deterioration were divided into 2 groups according to their outcome: weaned ECMO (W: n = 18) or not (NW: n = 7). In the W group, we examined the differences in parameters between the 2 time points, ECMO introduction, and the reduction in ECMO flow to 40% of the initial setting known as the conventional recovery point (C-point). Significant differences were observed in systolic pulmonary artery pressure, the cardiac index measured by the thermodilution method, C-reactive protein, lactate, base excess, and the end-tidal CO(2) concentration (ETCO(2)). Next, by closely examining these 6 parameters measured every 12 hours, we found that only ETCO(2) had always changed steeply, like a 'flexion point' (E-point), in all W cases, but not in NW. The E-point was defined as an initial increase in ETCO(2) of >or= 5 mmHg over the preceding 12 hours with a continued rise over the next 12 hours. E-points appeared as much as 95 +/- 60 hours earlier than C-points and also preceded weaning off of ECMO. ETCO(2) can be a useful continuous parameter for predicting the adequate timing of weaning off of ECMO for circulatory failure at the bedside.


title: [Cardiac arrest in severe acute respiratory syndrome: analysis of 15 cases].

text: OBJECTIVE To investigate the causes for cardiac arrest in severe acute respiratory syndrome (SARS) patients. METHODS Retrospective analysis of the epidemiological history, clinical presentation, the change of laboratory tests, chest radiography, and treatment of 15 SARS patients with cardiac arrest. RESULTS The average age of the patients was 60 years. Eight had a history of exposure to SARS patients, among them 6 were household contacts. Eight patients had no underlying diseases, and another 8 complained of extreme anxiety. Abnormalities of cardiac enzymes were present in 10 patients. Myocardial ischemia and arrhythmia were present in 5 patients. Bilateral, multifocal lung infiltrates were present in 13 of the 15 patients. Four patients died after defecation and 9 died during relatively stable periods. CONCLUSIONS It was suggested that the causes for cardiac arrest in SARS patients may include: (1) the lung injury caused by the SARS virus leads to hypoxemia and thus an unsteady state of the myocardial electricity; (2) SARS virus directly causes injury to the myocardial cells and/or the conduct system; (3) SARS infection aggravates the original myocardial pathological change, worsening the conduct block; (4) extreme anxiety leads to extra secretion of catecholamine, which causes instability of myocardial electricity; (5) defecation worsens hypoxemia, which induces arrhythmia (ventricular fibrillation) and causes cardiac arrest.


title: [ACE2 and coronavirus - a question of balance and dynamics?]

text: The new SARS-CoV-2 virus enters cells via angiotensin-converting enzyme 2 (ACE2). ACE2 counteracts ACE and angiotensin II in the renin-angiotensin-aldosterone system (RAAS) and has critical functions in the lung and cardiovascular system. SARS-CoV was found to down-regulate ACE2, leaving angiotensin II unbalanced in affected organs. A similar effect of SARS-CoV-2 could partly explain risk factors and symptoms, and could potentially be treatable.


Telepathic charm seduces audience at paranormal debate.


title: Human hepatic sinusoidal endothelial cells can be distinguished by expression of phenotypic markers related to their specialised functions in vivo.

text: The hepatic sinusoids are lined by a unique population of hepatic sinusoidal endothelial cells (HSEC), which is one of the first hepatic cell populations to come into contact with blood components. However, HSEC are not simply barrier cells that restrict the access of blood-borne compounds to the parenchyma. They are functionally specialised endothelial cells that have complex roles, including not only receptor-mediated clearance of endotoxin, bacteria and other compounds, but also the regulation of inflammation, leukocyte recruitment and host immune responses to pathogens. Thus understanding the differentiation and function of HSEC is critical for the elucidation of liver biology and pathophysiology. This article reviews methods for isolating and studying human hepatic endothelial cell populations using in vitro models. We also discuss the expression and functions of phenotypic markers, such as the presence of fenestrations and expression of VAP-1, Stabilin-1, L-SIGN, which can be used to identify sinusoidal endothelium and to permit discrimination from vascular and lymphatic endothelial cells.


title: Single-amino-acid substitutions in open reading frame (ORF) 1b-nsp14 and ORF 2a proteins of the coronavirus mouse hepatitis virus are attenuating in mice.

text: A reverse genetic system was recently established for the coronavirus mouse hepatitis virus strain A59 (MHV-A59), in which cDNA fragments of the RNA genome are assembled in vitro into a full-length genome cDNA, followed by electroporation of in vitro-transcribed genome RNA into cells with recovery of viable virus. The "in vitro-assembled" wild-type MHV-A59 virus (icMHV-A59) demonstrated replication identical to laboratory strains of MHV-A59 in tissue culture; however, icMHV-A59 was avirulent following intracranial inoculation of C57BL/6 mice. Sequencing of the cloned genome cDNA fragments identified two single-nucleotide mutations in cloned genome fragment F, encoding a Tyr6398His substitution in open reading frame (ORF) 1b p59-nsp14 and a Leu94Pro substitution in the ORF 2a 30-kDa protein. The mutations were repaired individually and together in recombinant viruses, all of which demonstrated wild-type replication in tissue culture. Following intracranial inoculation of mice, the viruses encoding Tyr6398His/Leu94Pro substitutions and the Tyr6398His substitution alone demonstrated log10 50% lethal dose (LD50) values too great to be measured. The Leu94Pro mutant virus had reduced but measurable log10 LD5), and the "corrected" Tyr6398/Leu94 virus had a log10 LD50 identical to wild-type MHV-A59. The experiments have defined residues in ORF 1b and ORF 2a that attenuate virus replication and virulence in mice but do not affect in vitro replication. The results suggest that these proteins serve roles in pathogenesis or virus survival in vivo distinct from functions in virus replication. The study also demonstrates the usefulness of the reverse genetic system to confirm the role of residues or proteins in coronavirus replication and pathogenesis.


title: Efficacy of beta-cyclodextrin against experimental cryptosporidiosis in neonatal lambs.

text: The efficacy of beta-cyclodextrin against experimental Cryptosporidium parvum infection was evaluated in neonatal lambs. The animals were treated by oral administration of the drug at 1 g/kg of body weight during 3 consecutive days. Preventive treatment was started within 1 day of birth, and therapeutic treatment was initiated at the onset of diarrhea following confirmation of infection. Disease development and drug efficacy were evaluated by monitoring the presence or absence of diarrhea and oocyst shedding from birth until 30 days of age. Weight gains at 15 and 30 days of age were also recorded. Beta-cyclodextrin was highly effective as a prophylactic treatment; 1 animal did not acquire the infection, diarrhea was prevented in infected animals, and there was a considerable decrease in oocyst shedding. The therapeutic treatment was effective in decreasing the severity of diarrhea and the duration of oocyst shedding. The animals tolerated the drug well, and there was a significant increase in their body weights.


title: Rangelia vitalii: changes in the enzymes ALT, CK and AST during the acute phase of experimental infection in dogs.

text: Rangelia vitalii is a protozoon that causes diseases in dogs, and anemia is the most common laboratory finding. However, few studies on the biochemical changes in dogs infected with this protozoon exist. Thus, this study aimed to investigate the biochemical changes in dogs experimentally infected with R. vitalii, during the acute phase of the infection. For this study, 12 female dogs (aged 6-12 months and weighing between 4 and 7 kg) were used, divided in two groups. Group A was composed of healthy dogs (n = 5); and group B consisted of infected animals (n = 7). Blood samples were collected on days 0, 10, 20 and 30 after infection, using tubes without anticoagulant to obtain serum and analyze the biochemical parameters. An increase in alanine aminotransferase (ALT) on day 20 (P < 0.05) was observed. Also, increased creatine kinase (CK) and aspartate aminotransferase (AST) levels were observed throughout the experimental period (P < 0.05). No changes in the serum gamma-glutamyltransferase, urea and creatinine levels were observed. Thus, is possible to conclude that experimental infection with R. vitalii in dogs causes changes to the biochemical profile, with increased ALT, AST and CK enzyme levels.


title: Prevalence of subclinical infection and transmission of severe acute respiratory syndrome (SARS) in a residential care home for the elderly.

text: OBJECTIVE To ascertain the prevalence of subclinical severe acute respiratory syndrome-coronavirus (SARS-CoV) infection and study the transmission of SARS-CoV in a local outbreak at a residential care home for the elderly. DESIGN Cross-sectional study. SETTING A residential care home for the elderly in Hong Kong with a local outbreak of SARS. PARTICIPANTS Residents and staff in the residential care home who had contact with three patients with SARS (residents A and B, and staff C). MAIN OUTCOME MEASURES Blood samples were tested for total antibodies to SARS-CoV by immunofluorescence antibody test. The transmission of SARS was elucidated based on information from standardised questionnaires, and records of investigation and surveillance by the Department of Health. RESULTS Among the 90 eligible residents, three died, one moved out, and 19 refused to participate. Of the 32 eligible staff, six refused to participate. None of the remaining 93 participants tested positive for antibody to SARS-CoV. Based on the chronological order, resident A might have transmitted infection to resident B and staff C. Sitting close to the bathroom doorway while resident A took a shower was the only contact of resident B with resident A. The only opportunity for staff C to have contact with body fluids/excreta of resident A was in the handling of rubbish from the resident's room. CONCLUSION Subclinical SARS-CoV infection was rare in a residential care home for the elderly with an outbreak of SARS. Nonetheless the close working and living conditions for staff and residents in such a home may facilitate transmission of SARS despite vigilant precautionary measures.


title: Prevalence of enteropathogenic Escherichia coli in naturally occurring cases of poult enteritis-mortality syndrome.

text: Enteropathogenic Escherichia coli (EPEC) previously were identified in poult enteritis-mortality syndrome (PEMS)-affected turkeys and associated as a cause of this disease. In the present study, the prevalence of EPEC in PEMS-affected turkeys was examined retrospectively with archived tissues and intestinal contents collected from 12 PEMS-affected turkey flocks in 1998. Formalin-fixed intestinal tissues were examined by light and electron microscopy for attaching and effacing (AE) lesions characteristic of EPEC, and frozen (-75 C) intestinal contents were examined for presence of EPEC. Escherichia coli isolates were characterized on the basis of epithelial cell attachment, fluorescent actin staining (FAS) test, and presence of E. coli attaching/effacing (EAE), shigalike toxin (SLT) type I, SLT II, and bundle-forming pilus (BFP) genes by polymerase chain reaction procedures. EPEC isolates were examined for pathogenicity and ability to induce AE lesions in experimentally inoculated young turkeys. AE lesions were identified by light microscopy in Giemsa-stained intestines from 7 of 12 PEMS-affected turkey flocks. Lesions consisted of bacterial microcolonies attached to epithelial surfaces with epithelial degeneration at sites of attachment and inflammatory infiltration of the lamina propria. Electron microscopy confirmed the identity of AE lesions in six of seven flocks determined to have AE lesions by light microscopy. EPEC were identified in 4 of 12 flocks on the basis of the presence of EAE genes a nd absence of SLT I and SLT II genes; all isolates lacked BFP genes. EPEC isolates produced AE lesions and variable mortality in turkeys coinfected with turkey coronavirus. In total, EPEC were associated with 10 of 12 (83%) naturally occurring PEMS cases on the basis of identification of AE lesions and/or EPEC isolates. These findings provide additional evidence suggesting a possible role for EPEC in the pathogenesis of PEMS.


title: A model for persistent murine coronavirus infection involving maintenance via cytopathically infected cell centres.

text: The relatively cell impermeable hygromycin B was found to inhibit viral but not cellular protein synthesis when added to cultures of murine hepatitis virus (MHV)-infected or mock-infected mouse L-2 fibroblasts. Membrane permeability, as judged by influx of sodium ions, has previously been demonstrated to be an MHV E2 glycoprotein-mediated, cytopathic effect of MHV infection in L-2 cells. It is therefore likely that the selective effect of hygromycin B on viral protein synthesis is a reflection of an increased drug penetration into virus-infected cells. Using hygromycin B as a marker for MHV-induced cell membrane cytopathology, the effects of drug treatment on a persistent MHV infection in mouse LM-K fibroblasts were investigated. MHV persistence in LM-K cells, which normally involves a steady state infection of 0.1 to 1% of the cells in culture, was found to be cured by hygromycin B treatment, as measured by the elimination of infectious virus from the supernatant medium. Hygromycin B also resulted in the eradication of MHV-specific RNA from LM-K cells, arguing against the presence of a non-cytopathically or latently infected subpopulation of cells.


title: 5'-proximal hot spot for an inducible positive-to-negative-strand template switch by coronavirus RNA-dependent RNA polymerase.

text: Coronaviruses have a positive-strand RNA genome and replicate through the use of a 3' nested set of subgenomic mRNAs each possessing a leader (65 to 90 nucleotides [nt] in length, depending on the viral species) identical to and derived from the genomic leader. One widely supported model for leader acquisition states that a template switch takes place during the generation of negative-strand antileader-containing templates used subsequently for subgenomic mRNA synthesis. In this process, the switch is largely driven by canonical heptameric donor sequences at intergenic sites on the genome that match an acceptor sequence at the 3' end of the genomic leader. With experimentally placed 22-nt-long donor sequences within a bovine coronavirus defective interfering (DI) RNA we have shown that matching sites occurring anywhere within a 65-nt-wide 5'-proximal genomic acceptor hot spot (nt 33 through 97) can be used for production of templates for subgenomic mRNA synthesis from the DI RNA. Here we report that with the same experimental approach, template switches can be induced in trans from an internal site in the DI RNA to the negative-strand antigenome of the helper virus. For these, a 3'-proximal 89-nt acceptor hot spot on the viral antigenome (nt 35 through 123), largely complementary to that described above, was found. Molecules resulting from these switches were not templates for subgenomic mRNA synthesis but, rather, ambisense chimeras potentially exceeding the viral genome in length. The results suggest the existence of a coronavirus 5'-proximal partially double-stranded template switch-facilitating structure of discrete width that contains both the viral genome and antigenome.