title: Infectious respiratory illnesses and their impact on healthcare workers: a review.

text: Respiratory illnesses are increasingly recognised as a growing concern for healthcare workers (HCWs) and patients. The recent hospital-based outbreak of Severe Acute Respiratory Syndrome (SARS) has once again highlighted the vulnerability of HCWs. The new epidemic of the 21st century resulted in tremendous economic and psychological impact with its high rates of mortality and nosocomial transmission. Even as the epidemic was brought under control within months, many details about the SARS coronavirus remained a mystery. The threat of another potential global outbreak continues to lurk in the background. Many valuable lessons have been learned through the SARS epidemic. It is, therefore, timely for us to review some of the respiratory pathogens that are well-known to cause nosocomial outbreaks. We need to be better armed to deal with future potential outbreaks and biohazardous situations. The importance of safeguarding the health of our medical staff and the community cannot be over-emphasised. In this paper, we review the incidence, transmission and various preventive strategies of respiratory illnesses in HCWs, in particular, new diagnostic tools, infection control management strategies, personal protective equipments, vaccination programmes and post-exposure prophylaxis.


BIS/BTS SARS guidelines.


Erratum to: High-Mannose Specific Lectin and Its Recombinants from a Carrageenophyta Kappaphycus alvarezii Represent a Potent Anti-HIV Activity Through High-Affinity Binding to the Viral Envelope Glycoprotein gp120.


Empiric Meropenem-based versus Ceftazidime-based Therapy for Severe Community-Acquired Pneumonia in a Retrospective Cohort Study.


title: B cells induce apoptosis via a novel mechanism in fibroblasts infected with mouse hepatitis virus.

text: B cells from nonimmune mice mediate the cytolysis of fibroblasts infected with the coronavirus, mouse hepatitis virus (MHV), strain A59. In this investigation, we report that splenic B cells and a B cell hybridoma induced the fragmentation of MHV-infected target cell DNA into a nucleosomal ladder pattern, characteristic of apoptosis. To determine the mechanism by which B cells mediated this killing event, we used criteria previously established for the killing of target cells by cytotoxic T lymphocytes (CTLs) and compared this B-cell-mediated killing to lymphocytic choriomeningitis virus (LCMV)-specific CTL killing of LCMV-infected target cells. Unlike CTL-mediated cytotoxicity, B cells efficiently lysed and induced the fragmentation of the DNA in their target cells in the presence of EGTA, arguing against a Ca(2+)-dependent granule exocytosis model for killing. In addition, paraformaldehyde-fixed B cells were able to kill MHV-infected targets. We were unable to detect TNF-alpha-associated cytotoxicity via bioassay with nonimmune effector B cells against the TNF-sensitive cell line, LM, or the TNF-alpha-resistant subline, L929.w, infected with MHV. Serine esterase inhibitors (benzamidine hydrochloride and N alpha-p-tosyl-L-arginine methyl ester) blocked CTL-induced 51Cr release and DNA fragmentation. In contrast, the inhibitors did not block the B-cell-induced 51Cr release, but did cause an inhibition in the fragmentation of the DNA of the target cell. These data indicate that B cells are capable of inducing the lysis and DNA fragmentation of MHV-infected target cells similar to CTL-induced apoptosis. However, we show that the mechanism(s) by which these processes are induced by B cells is distinct from CTL-mediated cytotoxicity.


title: Hepatitis C virus F protein sequence reveals a lack of functional constraints and a variable pattern of amino acid substitution.

text: Hepatitis C virus (HCV) is an important human pathogen that affects 170 million people worldwide. The HCV genome is an RNA molecule that is approximately 9.6 kb in length and encodes a polyprotein that is cleaved proteolytically to generate at least 10 mature viral proteins. Recently, a new HCV protein named F has been described, which is synthesized as a result of a ribosomal frameshift. Little is known about the biological properties of this protein, but the possibility that the F protein may participate in HCV morphology or replication has been raised. In this work, the presence of functional constraints in the F protein was investigated. It was found that the rate of amino acid substitutions along the F protein was significantly higher than the rate of synonymous substitutions, and comparisons involving genes that represented independent phylogenetic lineages yielded very different divergence/conservation patterns. The distribution of stop codons in the F protein across all HCV genotypes was also investigated; genotypes 2 and 3 were found to have more stop codons than genotype 1. The results of this work suggest strongly that the pattern of divergence in the F protein is not affected by functional constraints.


title: [Three years surveillance of viral etiology of acute lower respiratory tract infection in children from 2007 to 2010].

text: OBJECTIVE Viruses are common pathogens of acute lower respiratory tract infection (ALRTI) in children. There are few studies on consecutive monitoring of viral pathogens of ALRTI in a larger cohort during the past several years. The aim of this study was to investigate the viral pathogens of ALRTI in children of different age groups and to outline the epidemic feature of different viruses. METHOD (1) Totally 1914 (1281 male and 709 female) children with clinical diagnosis of ALRTI during the period of March 2007 to March 2010 were recruited into this study. These patients were hospitalized patients in department of internal medicine or outpatients in emergency department in Beijing Children's Hospital. The patients were divided into four groups, including 1072 patients < 1 year old, 326 patients 1- < 3 years old, 158 patients 3- < 6 years old, 358 patients ≥ 6 years old. One nasopharyngeal aspirate specimen was collected from each patient. Reverse transcription (RT) PCR methods were applied to detect common respiratory viruses including respiratory syncytial virus (RSV), human rhinovirus (HRV), influenza virus type A, B and C (IFA, IFB, IFC), parainfluenza virus (PIV) type 1-4, adenovirus (ADV), enterovirus (EV), human coronavirus (HCOV), human metapneumovirus (HMPV) and human bocavirus (HBOV). RESULT (1) The total positive rate of viruses was 70.3%. The positive rate was 83.0% (890/1072) in the group of < 1 year old, and 80.1% (261/326) in group of 1- < 3 years old, 60.8% (96/158) in group of 3- < 6 years old and 27.7% (99/358) in group of ≥ 6 years old, respectively. There was a significant difference in the positive rate among different age groups (χ² = 2213.5, P = 0.000). The top three viruses were RSV, HRV and PIV; and the positive rates were 50.9%, 36.2% and 12.0% respectively in group of < 1 year old. (2) The epidemic seasons of RSV and HRV were winter and spring, and PIV infection was epidemic in spring and summer. (3) The detection rates of 2 or more viruses were 38.2%, 36.4%, 30.2% and 15.2% in groups of < 1 year old, 1- < 3 years old, 3- < 6 years old and ≥ 6 years old, respectively. There was a significant difference in the mixed infection rate among different age groups (χ² = 1346.00, P = 0.000). CONCLUSION RSV, HRV and PIV were the most predominant pathogens in younger children with ALRTI. Different viral infections had different seasonal features. Mixed infections with two or more viruses were detected in substantial proportion of patients with ALRTI, but further studies are needed to explore the clinical significance of mixed infection with viruses in patients with ALRTI.


title: Chest radiograph scores as potential prognostic indicators in severe acute respiratory syndrome (SARS).

text: OBJECTIVE We analyzed serial chest radiographic scores for lung opacification in patients with severe acute respiratory syndrome (SARS) for temporal changes and differences between fatal and discharged cases. We sought to establish the earliest radiographic scores sensitive as potential prognostic indicators of fatal outcomes. MATERIALS AND METHODS Chest radiographs that had been obtained from presentation until the death or discharge of 313 patients with SARS were scored on the basis of the percentage area and location of lung opacification. Profile analysis and univariable logistic regression were performed on these radiographic scores. RESULTS Despite the increased mortality risks of advanced age and male sex, no significant difference was seen in the percentage area of opacification (AO%) between the sexes in either the group of patients with fatal outcomes or the group of patients who were discharged. No difference existed between age groups (< 65 years vs >/= 65 years), except for the radiograph showing the peak lung opacification in the deceased group in which the lungs of older patients had less opacification than those of younger patients. The radiographic scores obtained by day 7 were the earliest ones with good performance in prognostic prediction. The model showed good discriminatory performance, indicated by high C-indexes for receiver operator characteristic curves (0.86 for AO% and 0.90 for the number of opacified zones). The predicted proportion of patients with fatal outcomes showed high agreement with percentage of patients who died (goodness-of-fit statistic p = 0.18 for AO%, 0.73 for the number of opacified zones). By day 7, crude odds ratio of death was 1.73 per 5% of AO% (p < 0.0001) or 2.93 per lung zone opacified (p < 0.0001). CONCLUSION Chest radiographic scores (percentage of lung or the number of zones opacified) by day 7 could be used as fatal prognostic indicators.


title: Benefits of early haemofiltration during aorto-bifemural bypass with mesenteric revascularization - a case report.

text: The intraoperative vs postoperative initiation of haemofiltration procedures in patients submitted for major vascular surgery is a controversial issue and a subject of debate in recent literature. We report the case of a 50 yr old patient scheduled for aorto-bifemoral bypass with mesenteric revascularization in whom the haemofiltration procedure (Prismaflex with Onix filter) was installed intraoperatively. Known to have non-insulin-dependent type 2 diabetes, the patient was admitted for Leriche syndrome, abdominal aorta thrombosis, superior and inferior mesenteric artery occlusion, celiac trunk occlusion, bilateral critical limb ischemia and mild renal impairment. The filtration rate was 25 ml/kg/h, ultrafiltration rate of 50 ml/h and 2 h clampation time. Haemofiltration was continued postoperatively in the ICU for another 48 h. The patient had a favorable evolution with restoration of renal function and a significant improvement of the biochemical parameters. In conclusion the early haemofiltration applied in this case provided clear beneficial effects, probably preventing the evolution towards multiple organ dysfunction syndrome.


title: Laparoscopic distal pancreatectomy preserving the spleen and splenic vessels for benign and low-grade malignant pancreatic neoplasm.

text: BACKGROUND/AIMS Laparoscopic spleen-preserving distal pancreatectomy (LSPDP) is expected to be less invasive than laparoscopic distal pancreatectomy with splenectomy. However, there are few reports regarding the details of the procedure for LSPDP, and its safety remains unclear. This study aimed to evaluate the feasibility and safety of LSPDP. METHODOLOGY Six patients underwent LSPDP from March 2009 to February 2013 in our center, and their clinical data and outcomes were reviewed retrospectively. RESULTS A total of six laparoscopic distal pancreatic resections were attempted in four female and two male patients. All of the operations were successful, with an average operative time of 290.7 min (range: 211-377 min) and an average blood loss of 43.5 g (range: 0-142 g). The mean hospital stay was 11.8 days (range: 9-17days). No obvious pancreatic fistulas occurred, although pseudocysts at the stump of the pancreas were recognized in three patients on CT scans performed at 7 days postoperatively. Postoperative pathological examinations revealed two cases of serous cystadenoma in the body and tail of the pancreas, one case of serous oligocystic adenoma, one case of mucinous cystadenoma, one case of neuroendocrine tumor, and one case of solid-pseudopapillary neoplasm. CONCLUSIONS LSPDP is minimally invasive, safe, and feasible for the management of benign pancreatic tail tumors, with the advantages of earlier recovery and less morbidity from complications.


title: New DNA viruses identified in patients with acute viral infection syndrome.

text: A sequence-independent PCR amplification method was used to identify viral nucleic acids in the plasma samples of 25 individuals presenting with symptoms of acute viral infection following high-risk behavior for human immunodeficiency virus type 1 transmission. GB virus C/hepatitis G virus was identified in three individuals and hepatitis B virus in one individual. Three previously undescribed DNA viruses were also detected, a parvovirus and two viruses related to TT virus (TTV). Nucleic acids in human plasma that were distantly related to bacterial sequences or with no detectable similarities to known sequences were also found. Nearly complete viral genome sequencing and phylogenetic analysis confirmed the presence of a new parvovirus distinct from known human and animal parvoviruses and of two related TTV-like viruses highly divergent from both the TTV and TTV-like minivirus groups. The detection of two previously undescribed viral species in a small group of individuals presenting acute viral syndrome with unknown etiology indicates that a rich yield of new human viruses may be readily identifiable using simple methods of sequence-independent nucleic acid amplification and limited sequencing.


title: Analysis in vivo of turnip crinkle virus satellite RNA C variants with mutations in the 3'-terminal minus-strand promoter.

text: Turnip crinkle virus and its associated RNA, sat-RNA C, share similar, but not identical hairpins near their 3' ends and terminate with CCUGCCC-OH, which forms a single-stranded tail. With an in vitro transcription system containing partially purified TCV RdRp, the 3'-terminal 29 bases making up the hairpin and single-stranded tail were previously demonstrated to be required for transcription, and alterations in the stem, but not the loop, could affect template activity (C. Song and A. E. Simon, 1995, J. Mol. Biol. 254, 6-14). We have now analyzed sat-RNA C mutants in the 3' hairpin for ability to accumulate in vivo. While active templates in vitro were able to accumulate in vivo, some very weak templates in vitro were also able to accumulate in vivo without reversion or second-site alterations. Computer models of hairpin structure indicated that biologically active promoters could have hairpins less stable than wild type, with loops of variable length and sequence, and without a need for a 6-base single-stranded tail. In addition, transcripts containing compensatory exchanges in the upper stem region that had limited activity in vitro were biologically active in vivo, indicating that positioning of specific bases in the stem is not required to produce an active minus-strand promoter.


title: Current status of medical treatment for abdominal aortic aneurysm.

text: Although cardiovascular disease is widely recognized as the leading cause of death, a lesser known fact is that aortic aneurysm is the 15th leading cause of death over the age of 65 years in the USA. The golden standard of the treatments are invasive interventions either with open surgical repair (OS) or endovascular aneurysm repair (EVAR). The concept of medical treatment is to prevent abdominal aortic aneurysm (AAA) from rupture and avoid surgical treatment by preventing aneurysm enlargement or even reducing aneurysm size. Matrix metalloproteinases (MMP) are structurally related metalloendopeptidases that can degrade the extracellular matrix and is thought to play important roles in AAA. There are many proposed pharmacological treatments including: β-blockers, angiotensin-converting enzyme inhibitor (ACE inhibitors), angiotensin-receptor blocker (ARB), statins, macrolides and, doxycycline, an inhibitor of the MMP. The latter is a potential promising drug as medical treatment for AAA and the Non-invasive Treatment of Abdominal Aortic Aneurysm Clinical Trial (N-TA(3)CT) is currently ongoing in the USA. Here, the pathophysiology and potential medical therapy for AAA will be reviewed.


title: SARS and emerging infectious diseases: a challenge to place global solidarity above national sovereignty.

text: Severe acute respiratory syndrome (SARS) emerged in a world where information about infectious disease outbreaks travels at speeds and in ways not imagined just 30 years ago, and where scientists are increasingly working together on detecting and responding to public health events that threaten international public health and economic security. The SARS outbreak clearly demonstrated that it is no longer the exclusive privilege of countries to report and respond to infectious diseases occurring in their own territories, but that the global community has also assumed this role, aided by the ease and power of electronic communication through the World Wide Web. This phenomenon has been cited by some scholars as a potential infringement on national sovereignty that compromises the concept that states reign supreme over their territories and peoples. At the same time, however, countries are increasingly seeking to collaborate internationally in infectious disease surveillance and response, as shown in the current situation of avian influenza (H5N1), and in the formal agreement leading to the revised International Health Regulations (IHR), suggesting that a new world order prevails over issues that once had been considered the sole domain of a sovereign nation.


title: Towards powerful experimental and statistical approaches to study intraindividual variability in labile traits.

text: There is a long-standing interest in behavioural ecology, exploring the causes and correlates of consistent individual differences in mean behavioural traits ('personality') and the response to the environment ('plasticity'). Recently, it has been observed that individuals also consistently differ in their residual intraindividual variability (rIIV). This variation will probably have broad biological and methodological implications to the study of trait variation in labile traits, such as behaviour and physiology, though we currently need studies to quantify variation in rIIV, using more standardized and powerful methodology. Focusing on activity rates in guppies (Poecilia reticulata), we provide a model example, from sampling design to data analysis, in how to quantify rIIV in labile traits. Building on the doubly hierarchical generalized linear model recently used to quantify individual differences in rIIV, we extend the model to evaluate the covariance between individual mean values and their rIIV. After accounting for time-related change in behaviour, our guppies substantially differed in rIIV, and it was the active individuals that tended to be more consistent (lower rIIV). We provide annotated data analysis code to implement these complex models, and discuss how to further generalize the model to evaluate covariances with other aspects of phenotypic variation.


title: Induction of mucosal immunity in cotton rats to haemagglutinin-esterase glycoprotein of bovine coronavirus by recombinant adenovirus.

text: An effective vaccine against enteric bovine coronavirus (BCV) must be able to induce mucosal immunity. We recently described the construction of recombinant human adenovirus type 5 (hAd5) carrying the BCV haemagglutinin-esterase (HE) gene in the early transcription region 3 of the adenovirus genome. In this study, we examined the induction of systemic and mucosal immune responses to the hAd5 vector carrying the BCV HE gene (AdBcHE) following intranasal or enteric immunization of cotton rats. Regardless of the route of administration, mucosal immunization with AdBcHE induced significant levels of anti-HE IgG antibodies in serum. In addition, following intranasal immunization with AdBcHE, significant levels of anti-HE IgA antibodies were found in lung washes of immunized cotton rats. Furthermore, the specific anti-HE antibodies in sera and mucosal secretions efficiently neutralized BCV infectivity in vitro. T-cell proliferation and cell-mediated cytotoxic responses against the BCV HE were elicited in the spleen of intranasally immunized animals. The results demonstrate that mucosal immunization with AdBcHE is capable of inducing both systemic and mucosal immunity to the BCV HE. These immune responses may be important in protecting animals from BCV infection.


title: The spike glycoprotein of murine coronavirus MHV-JHM mediates receptor-independent infection and spread in the central nervous systems of Ceacam1a-/- Mice.

text: The MHV-JHM strain of the murine coronavirus mouse hepatitis virus is much more neurovirulent than the MHV-A59 strain, although both strains use murine CEACAM1a (mCEACAM1a) as the receptor to infect murine cells. We previously showed that Ceacam1a(-/-) mice are completely resistant to MHV-A59 infection (E. Hemmila et al., J. Virol. 78:10156-10165, 2004). In vitro, MHV-JHM, but not MHV-A59, can spread from infected murine cells to cells that lack mCEACAM1a, a phenomenon called receptor-independent spread. To determine whether MHV-JHM could infect and spread in the brain independent of mCEACAM1a, we inoculated Ceacam1a(-/-) mice. Although Ceacam1a(-/-) mice were completely resistant to i.c. inoculation with 10(6) PFU of recombinant wild-type MHV-A59 (RA59) virus, these mice were killed by recombinant MHV-JHM (RJHM) and a chimeric virus containing the spike of MHV-JHM in the MHV-A59 genome (SJHM/RA59). Immunohistochemistry showed that RJHM and SJHM/RA59 infected all neural cell types and induced severe microgliosis in both Ceacam1a(-/-) and wild-type mice. For RJHM, the 50% lethal dose (LD(50)) is <10(1.3) in wild-type mice and 10(3.1) in Ceacam1a(-/-) mice. For SJHM/RA59, the LD(50) is <10(1.3) in wild-type mice and 10(3.6) in Ceacam1a(-/-) mice. This study shows that infection and spread of MHV-JHM in the brain are dependent upon the viral spike glycoprotein. RJHM can initiate infection in the brains of Ceacam1a(-/-) mice, but expression of mCEACAM1a increases susceptibility to infection. The spread of infection in the brain is mCEACAM1a independent. Thus, the ability of the MHV-JHM spike to mediate mCEACAM1a-independent spread in the brain is likely an important factor in the severe neurovirulence of MHV-JHM in wild-type mice.


title: Systematic Review of the BridgePoint System for Crossing Coronary and Peripheral Chronic Total Occlusions.

text: BACKGROUND The BridgePoint system consists of the CrossBoss coronary catheter and Stingray CTO system (Boston Scientific) for coronary chronic total occlusions (CTOs), and the Viance crossing catheter and Enteer re-entry system (Covidien) for peripheral CTOs. METHODS We performed a systematic review of the literature on the BridgePoint system published between October 2008 and August 2014, in accordance with the standards set forth in the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. RESULTS We identified a total of 20 studies: 12 studies on coronary CTOs (10 retrospective with 2 prospective case series) and 8 studies on peripheral CTOs (all retrospective). Among 320 patients undergoing coronary CTO intervention, pooled technical success and major adverse cardiac event rates were 77.1% and 3.8%, respectively. Among 175 patients undergoing peripheral CTO intervention, pooled technical success and major adverse events were 82.2% and 4.6%, respectively. Only 1 study reported long-term outcomes after use of the coronary BridgePoint system, demonstrating similar incidence of major adverse cardiac events between BridgePoint-treated and non-BridgePoint treated patients; however, the study had low power. CONCLUSIONS The BridgePoint system is associated with high procedural success rates and low complication rates when used for crossing both coronary and peripheral CTOs.


title: Reproduction of PMWS in immunostimulated SPF piglets transfected with infectious cloned genomic DNA of type 2 porcine circovirus.

text: Postweaning multisystemic wasting syndrome (PMWS) is a recently emerged disease affecting pigs. Type 2 porcine circovirus (PCV2) has been associated with this syndrome although other factors are required in association with this virus for PMWS expression. The aim of this study was to investigate whether general immunostimulation (injections of keyhole limpet hemocyanin emulsified in incomplete Freund adjuvant and of thioglycollate medium) could strengthen the severity of PMWS in six-week-old specific-pathogen-free (SPF) piglets transfected with pure tandem-cloned PCV2 DNA by the intramuscular route. Non-immunostimulated piglets transfected with the viral clone did not present clinical signs but only mild pathological microlesions characteristic of PMWS. These piglets seroconverted and high viral genome loads and infectious titers were detected in the lymphoid organs at the end of the trial. Mild-to-moderate forms of PMWS were generally observed in the immunostimulated transfected piglets, as well as one severe form for a piglet (8003) which died. These piglets with mild-to-moderate forms had higher DNA loads than the transfected-only animals. Thus, viral replication was enhanced by immunostimulation. This is the first time that clinical PMWS has been reported in an SPF immunostimulated piglet infected with a pure inoculum consisting of tandem-cloned PCV2 DNA. This result confirms that PCV2 is the agent of PMWS and that immunostimulation could enhance PMWS in SPF piglets transfected with a PCV2 DNA clone.


title: Severe acute respiratory syndrome: report of treatment and outcome after a major outbreak.

text: BACKGROUND The outcome is reported of a prospective uncontrolled study based on a stepwise treatment protocol during an outbreak of severe acute respiratory syndrome (SARS) in Hong Kong. METHOD One hundred and thirty eight patients were treated with broad spectrum antibiotics, a combination of ribavirin and low dose corticosteroid, and then intravenous high dose methylprednisolone according to responses. Sustained response to treatment was defined as (1) defervescence for > or =4 consecutive days, (2) resolution of lung consolidation by >25%, and (3) oxygen independence by the fourth day without fever. Patients with defervescence who achieved either criterion 2 or 3 were classified as partial responders. Patients who fell short of criteria 2 and 3 were non-responders. RESULTS Laboratory confirmation of SARS coronavirus infection was established in 132 (95.7%). None responded to antibiotics but 25 (18.1%) responded to ribavirin + low dose corticosteroid. Methylprednisolone was used in 107 patients, of whom 95 (88.8%) responded favourably. Evidence of haemolytic anaemia was observed in 49 (36%). A high level of C-reactive protein at presentation was the only independent predictor for use of methylprednisolone (odds ratio 2.18 per 10 mg/dl increase, 95% confidence interval 1.12 to 4.25, p = 0.02). Thirty seven patients (26.8%) required admission to the intensive care unit and 21 (15.2%) required invasive mechanical ventilation. There were 15 deaths (mortality rate 10.9%), most with significant co-morbidities, whereas 122 (88.4%) had been discharged home 4 months after the outbreak onset. CONCLUSION The use of high dose pulse methylprednisolone during the clinical course of a SARS outbreak was associated with clinical improvement, but randomised controlled trials are needed to ascertain its efficacy in this condition.


title: A study on SARS awareness and health-seeking behaviour - findings from a sampled population attending National Healthcare Group Polyclinics.

text: INTRODUCTION The study aimed to assess the effectiveness of massive SARS public education effort on SARS awareness and the conduct of those suspected of having SARS. MATERIALS AND METHODS Five hundred and ninety-three respondents attending the National Healthcare Group Polyclinics (NHGP) participated in the survey from 9 to 13 June 2003. Associations between awareness of SARS symptoms and (i) first action to be taken and (ii) mode of transportation used, if the respondent was suspected of having SARS, were analysed using Chi-square or Fisher's exact tests. Logistic regression was performed to adjust for relevant covariates. RESULTS The majority (92.7%) of the respondents were aware of SARS symptoms. Television (91.6%), newspaper (65.2%) and radio (30.4%) formed the top 3 sources of information on SARS. Slightly more than half (51.6%) of those who suspect themselves of having SARS would choose to visit their primary health care doctors, while 22.7% of the respondents would go to Tan Tock Seng Hospital (TTSH). If they suspected themselves to have SARS, most (84.9%) of the 578 respondents would react appropriately by taking the SARS ambulance or driving themselves to TTSH. However, 60 respondents would nonetheless take public transport to TTSH [by taxi 8.5%, mass rapid transit (MRT) or bus 1.9%]. In particular, the retired with lower educational levels were likely to be oblivious both to the symptoms of SARS and the possible consequences of travelling by inappropriate transport. CONCLUSION Despite more than 2 months of intensive SARS public education in Singapore, there remain important gaps in knowledge and appropriate behaviour that have to be bridged.


title: Trajectory of Post-operative Wound Pain within the First Two Weeks following Posterior Spinal Fusion Surgery in Adolescent Idiopathic Scoliosis Patients.

text: STUDY DESIGN Prospective Cohort Study. OBJECTIVE To determine and evaluate the trajectory of surgical wound pain from day 1 to day 14 after PSF surgery in AIS patients. SUMMARY OF BACKGROUND DATA Information regarding how the post-operative pain improves with time offers invaluable information not only to the patients and parents but also to assist the clinician in managing post-operative pain. METHODS AIS patients who were planned for elective PSF surgery from September 2015 to December 2015 were prospectively recruited into this study. All patients underwent a similar pain management regime with patient controlled anesthesia (PCA) morphine, acetaminophen, celecoxib, oxycodone hydrochloride. RESULTS A total of 40 patients (36F:4 M) were recruited. The visual analogue score (VAS) pain score was highest at 12 hours post-operation (6.0 ± 2.3). It reduced to 3.9 ± 2.2 (day 4), 1.9 ± 1.6 (day 7) and 0.7 ± 1.1 (day 14). The total PCA usage in all patients was 12.4 ± 9.9 mg (1 12 hours), 7.1 ± 8.0 mg (12 to 24 hours), 5.6 ± 6.9 (24 to 36 hours) and 2.1 ± 6.1 mg (36 to 48 hours). The celecoxib capsules usage was reducing from 215.0 ± 152.8 mg at 24 hours to 55.0 ± 90.4 mg on day 14. The acetaminophen usage was reducing from 2275 ± 1198 mg at 24 hours to 150 ± 483 mg at day 14. Oxycodone hydrochloride capsules consumption rose to the peak of 1.4 ± 2.8 mg on day 4 before gradually reducing to none by day 13. CONCLUSIONS With an adequate post-operation pain regime, significant pain should subside to a tolerable level by post-operative day 4 and negligible by post-operative day 7. Patient usually can be discharged on post-operative day 4 when the usage of PCA morphine was not required.


title: The AAUAAA motif of bamboo mosaic virus RNA is involved in minus-strand RNA synthesis and plus-strand RNA polyadenylation.

text: Bamboo mosaic virus (BaMV) has a single-stranded positive-sense RNA genome with a 5'-cap structure and a 3' poly(A) tail. Deleting the internal loop that contains the putative polyadenylation signal (AAUAAA) in the 3' untranslated region (UTR) of BaMV genomic RNA appeared to diminish coat protein accumulation to 2% (C. P. Cheng and C. H. Tsai, J. Mol. Biol. 288:555-565, 1999). To investigate the function of the AAUAAA motif, mutations were introduced into an infectious BaMV cDNA at each residue except the first nucleotide. After transfection of Nicotiana benthamiana protoplasts with RNA transcript, the accumulations of viral coat protein and RNAs were determined. Based on the results, three different categories could be deduced for the mutants. Category 1 includes two mutants expressing levels of the viral products similar to those of the wild-type virus. Six mutations in category 2 led to decreased to similar levels of both minus-strand and genomic RNAs. Category 3 includes the remaining seven mutations that also bring about decreases in both minus- and plus-strand RNA levels, with more significant effects on genomic RNA accumulation. Mutant transcripts from each category were used to infect N. benthamiana plants, from which viral particles were isolated. The genomic RNAs of mutants in category 3 were found to have shorter poly(A) tails. Taken together, the results suggest that the AAUAAA motif in the 3' UTR of BaMV genomic RNA is involved not only in the formation of the poly(A) tail of the plus-strand RNA, but also in minus-strand RNA synthesis.


title: Increased susceptibility to septic and endotoxic shock in monocyte chemoattractant protein 1/cc chemokine ligand 2-deficient mice correlates with reduced interleukin 10 and enhanced macrophage migration inhibitory factor production.

text: The chemokine monocyte chemoattractant protein 1/CC chemokine ligand 2 (MCP-1/CCL2) is a potent chemoattractant of mononuclear cells and a regulatory mediator involved in a variety of inflammatory diseases. In the present study, we demonstrate that mcp-1/ccl2-deficient mice are more susceptible to systemic inflammatory response syndrome induced by lipopolysaccharide and to polymicrobial sepsis induced by cecum ligation and puncture (CLP) when compared with wild-type mice. Interestingly, in the CLP model, mcp-1/ccl2-deficient mice efficiently cleared the bacteria despite an impaired recruitment of leukocytes, especially mononuclear cells. The increased lethality rate in these models correlates with an impaired production of interleukin (IL) 10 in vivo. Furthermore, macrophages from mcp-1/ccl2-deficient mice activated with lipopolysaccharide also produced lower amounts of IL-10 and similar tumor necrosis factor compared with wild-type mice. We observed a drastic increase in the amounts of macrophage migration inhibitory factor at 6 and 24 h after CLP in mcp-1/ccl2-deficient mice. These results indicate that endogenous MCP-1/CCL2 positively regulates IL-10 but negatively controls macrophage migration inhibitory factor during peritoneal sepsis, thus suggesting an important immunomodulatory role for MCP-1/CCL2 in controlling the balance between proinflammatory and anti-inflammatory factors in sepsis.


title: [Clinical aspects of human infection by the avian influenza virus].

text: The species barrier is not perfect for Influenza A and numerous transmissions of the virus from pigs or poultry to humans have been described these years. Appearing in 1997 and becoming epidemic in 2003, influenza A/H5N1 provoked many deadly enzootics in poultry batteries (highly pathogenic avian influenza of HPAI). Starting in Asia, many countries throughout Africa and Europe were affected. Sporadic human cases were described in direct contact with diseased chicken or other poultry. Half of the cases are lethal, but human to human transmission occurs with difficulty. From January 2003 to August 11th 2009, 438 cases were declared worldwide with 262 deaths. Many countries declared cases, but recently most cases occurred in Egypt. Measures in hospital were taken which were copied from the measures for SARS (Severe Acute Respiratory Syndrome), but these were probably excessive in this case, considering the low rate of secondary cases with A/H5N1. In many human infections, signs of severe respiratory distress develop and multi organ failure. It was feared that this deadly virus could become easily transmitted between humans, leading to a new pandemic. This was not the case up to now. The strong pathogenicity of the virus is still not completely explained, but the deep location of infection in the lungs and the deregulation of cytokine production by the target cells, particularly macrophages, may be part of the explanation.


title: Comparison of the spike precursor sequences of coronavirus IBV strains M41 and 6/82 with that of IBV Beaudette.

text: The nucleotide sequences of the spike precursor genes of infectious bronchitis virus strains M41 and 6/82 have been determined and compared with that of the Beaudette strain which we have previously sequenced. The two Massachusetts strains, M41 and Beaudette, were found to be remarkably similar, having only 3.7% of the amino acids different. The situation with 6/82, one of the new field isolates, is quite different and this strain had 13.8% of its amino acids different from Beaudette. The differences identified are discussed in terms of the structural features of the spike protein.


title: Identification of a newly isolated avian infectious bronchitis coronavirus variant in China exhibiting affinity for the respiratory tract.

text: Twelve infectious bronchitis virus (IBV) isolates obtained from commercial chickens in China between 2005 and 2006 were characterized by reverse transcriptase-polymerase chain reaction (RT-PCR) and the sequencing of the entire S1 gene. CK/CH/LSD/05I--an IBV variant, which was unlike the nephropathogenic IBV isolates found in China--exhibited an affinity for the respiratory tract. The variant was identified by phylogenic analysis and basic local alignment search tool (BLAST) searches of the entire S1 gene and by the vaccination-challenge test that was performed using heterologous strains. Further, it was demonstrated that the commercially used H120 vaccine did not provide sufficient protection against this variant; however, the attenuated heterologous IBV tl/CH/LDT3/03 P120, whose parent virus was isolated in China, showed a better efficacy of protection against CK/CH/LSD/05I. This study thus may demonstrate that the use of a combination of commercially available vaccines or of attenuated heterologous strains would provide satisfactory protection against the variant CK/CH/LSD/05I. In addition, the study also revealed that IBV strains exhibiting different pathogenicities were found cocirculating in the chicken flock in China.


title: Spleen preserving laparoscopic distal pancreatectomy for treatment of pancreatic lesions.

text: AIM Aim of this study is to evaluate the feasibility and safety of the laparoscopic approach in the treatment of distal pancreas tumors, from prospectively collected data. MATERIAL OF STUDY From January 2003 to July 2013, 20 patients were treated by laparoscopic approach for distal pancreatic lesions. Nine patients underwent laparoscopic pancreatic tumorectomy (LPT) (Group A) for insulinoma (mean lesion diameter 1.2 cm, range, 0.5-2) and 11 patients underwent spleen preserving laparoscopic distal pancreatectomy (SP-LDP) (Group B) for ductal adenocarcinoma (pT1N0R0) (1), cystic mucinous neoplasm (5), serous cystadenoma (4) and lymphoepithelial cysts (1). RESULTS Mean operative time was 94.3 minutes (range 80-110) for Group A and 164 minutes (range 90-240) for Group B. Intraoperative bleeding occurred in 4 cases (20%) and was easily controlled by laparoscopy. Conversion to open surgery was not required in any case. Morbidity was observed in 2 patients (18%) in Group A: pancreatic fistula (1) and peritoneal fluid collection (1); and a peritoneal fluid collection occurred in one patients (11%) in Group B. Mean hospital stay was 6.8 days (range 3-11) in Group A and 6.5 days (range 3-10) in Group B. Mortality was nil. At a mean follow-up of 82 months (range 15-141) local recurrence and distant metastases were not observed. DISCUSSION LDP is a valid treatment showing the same rate of complication to open surgery but allowing the advantages of a minimally invasive procedure. CONCLUSIONS SP-LDP is feasible and safe for benign and malignant pancreatic lesions.


Explosion of internet advertisements for protection against SARS.


Irreversible inhibitors of serine, cysteine, and threonine proteases.


title: [Characteristics of peripheral blood leukocyte differential counts in patients with COVID-19].

text: To investigate the early changes of peripheral blood leukocyte differential counts in patients with COVID-19. Ten patients with COVID-19 and 30 patients with other viral pneumonia (non-COVID-19) admitted to Shanghai Jiao Tong University Affiliated Sixth People's Hospital and Jinshan Branch Hospital from January 22 to February 17, 2020 were enrolled in this study. The differential counts of white blood cells were analyzed. Patients in COVID-19 group showed relatively lower absolute white blood cell (WBC) count 4.95(3.90,6.03)×10(9)/L, lymphocyte absolute count 1.20(0.98,1.50)×10(9)/L and eosinophil absolute count 0.01(0.01,0.01)×10(9)/L. Leukopenia developed in two patients(2/10), lymphocytopenia also in two patients(2/10). Seven over ten patients presented with eosinophil cytopenia. In non-COVID-19 group, absolute WBC count was 8.20 (6.78,9.03) ×10(9)/L (P<0.001), lymphocyte absolute count 1.75(1.20,2.53)×10(9)/L(P=0.036), eosinophil absolute count 0.02(0.01,0.03)×10(9)/L(P=0.05). Lymphocytopenia occurred in (16.7%) patients, eosinophil cytopenia in 16.7% patients too. In conclusion, leukopenia, lymphocytopenia and eosinophil cytopenia are more common in COVID-19 patients than those in non- COVID-19 patients.


Class - the ignored determinant of the nation's health.


title: [Coronavirus Covid-19 infection and breastfeeding: an exploratory review].

text: BACKGROUND The new coronavirus disease is an infectious disease caused by the SARS-Cov-2 virus, considered by the World Health Organization (WHO) an international public health emergency that may have negative consequences during breastfeeding. The objective of this work is to investigate the action plan on breastfeeding in postpartum women with SARS-CoV-2 and her newborn. METHODS A literature search has been conducted through the Medline, Web of Science, Scopus, BVS, and Cuiden databases. The methodological quality of the articles has been assessed using the "Grading of Recommendations Assessment, Development and Evaluation" (GRADE). This study has not been registered in PROSPERO. RESULTS A total of 14 documents have been found, of which 9 are observational empirical studies. Most of the studies were conducted in China, Italy, the USA, and Australia. A total of 114 mothers infected with coronavirus with their respective newborns have been assessed. The analyzed investigations state that it is best for the newborn to be breastfed; given that mother's milk samples were analyzed, detecting the presence of antibodies of the coronavirus in them, being a protective factor against infection. CONCLUSIONS Breastfeeding in postpartum women with SARS-CoV-2 is highly recommended for the newborn, if the health of the mother and newborn allow it. When direct breastfeeding is favoured, the appropriate respiratory hygiene measures always have to be considered. Whether the mother's health does not permit direct breastfeeding, her breast milk should be previously extracted and kept unpasteurized. To secure the newborn feeding, milk banks are also an appropriate option.


title: Mice transgenic for human angiotensin-converting enzyme 2 provide a model for SARS coronavirus infection.

text: To establish a small animal model of severe acute respiratory syndrome (SARS), we developed a mouse model of human severe acute respiratory syndrome coronavirus (SARS-CoV) infection by introducing the human gene for angiotensin-converting enzyme 2 (hACE2) (the cellular receptor of SARS-CoV), driven by the mouse ACE2 promoter, into the mouse genome. The hACE2 gene was expressed in lung, heart, kidney, and intestine. We also evaluated the responses of wild-type and transgenic mice to SARS-CoV inoculation. At days 3 and 7 postinoculation, SARS-CoV replicated more efficiently in the lungs of transgenic mice than in those of wild-type mice. In addition, transgenic mice had more severe pulmonary lesions, including interstitial hyperemia and hemorrhage, monocytic and lymphocytic infiltration, protein exudation, and alveolar epithelial cell proliferation and desquamation. Other pathologic changes, including vasculitis, degeneration, and necrosis, were found in the extrapulmonary organs of transgenic mice, and viral antigen was found in brain. Therefore, transgenic mice were more susceptible to SARS-CoV than were wild-type mice, and susceptibility was associated with severe pathologic changes that resembled human SARS infection. These mice will be valuable for testing potential vaccine and antiviral drug therapies and for furthering our understanding of SARS pathogenesis.


title: Serological studies on the potential synergism of porcine reproductive and respiratory syndrome virus and influenza-, corona- and paramyxoviruses in the induction of respiratory symptoms in swine.

text: Sera from 265 finishing pigs belonging to 70 herds, in which severe respiratory disorders were observed, were examined for antibody prevalence to porcine reproductive and respiratory syndrome virus (PRRSV), influenza virus subtypes H3N2 and H1N1, porcine respiratory corona virus (PRCV) and a recently described porcine paramyxovirus (PPMV). By immunoperoxidase-monolayer assay 69.1% of these sera were positive for PRRSV. Hemagglutination inhibiting activity was found in 55.1% of the sera for influenza virus subtype H1N1 (strain A/swine/Arnsberg/1/81). in 51.3% for influenza virus subtype H3N2 (strain A/Hong Kong/1/68) and in 14.3% for PPMV. PRCV specific antibodies, as determined by differential competitive blocking enzyme-linked immunosorbent assay were demonstrated in 192 of 236 (81.3%) sera. In order to reveal associations interspecific coefficients were calculated for antibody prevalence between PRRSV and the other viruses. Positive associations to PRRSV titres were found to PRCV, PPMV and influenza virus subtype H1N1 titres. chi-square analysis showed the statistical significance of associations regarding PRCV and influenza virus subtype H1N1. Depletion of lung macrophages after PRRSV infection is discussed as a possible mechanism for the promotion of secondary infections.


title: [Management of a colon cancer patient complicated with COVID-19].

text: OBJECTIVE To explore the feasibility of radical resection for cancer patients complicated with coronavirus disease 2019 (COVID-19). METHODS The management and clinical outcome of a sigmoid cancer patient with COVID-19 were analyzed. RESULTS The inflammation indicators and fever of this patient were effectively controlled and the lung lesions remained stable after active anti-viral treatment, then the radical colorectomy was performed after the viral negative conversion for twice. CONCLUSIONS The case indicates that radical resection can be performed in SARS-CoV-2 patients with twice-negative SARS-CoV-2 nucleic acid testing results.


title: [Lack of clinical evidence for the use of hydroxychloroquine to treat SARS-CoV-2 infection].

text: The severity of COVID-19 disease has led to an urgent need for the discovery of new treatments. Thus, global stocks of hydroxychloroquine (HCQ) have been put under pressure with a study of 26 patients treated with HCQ during their infection with SARS-CoV-2. Despite the study's lack of quality, several countries' medicines agencies subsequently issued guidelines for the use of HCQ for COVID-19. This review aims to elucidate potential mechanisms, which make HCQ treatment interesting in the fight against SARS-CoV-2 infection, as well as the current evidence for clinical use of HCQ to treat COVID-19.


Who should be immunised against hepatitis B?


title: Development, implementation and outcomes of a training program for responders to acts of terrorism.

text: INTRODUCTION Responding to acts of terrorism requires the effective use of public-safety and medical-response resources. The knowledge, skills and attitudes necessary to respond to future threats is unfamiliar to most emergency responders. OBJECTIVES The purpose of this report to describe the development, implementation and evaluation of a multidisciplinary, interactive and simulation-enhanced course to prepare responders to acts of terrorism. METHODS We used a 5-step systematic process to develop a blended-learning, simulation-enhanced training program. Learners completed a self-confidence questionnaire and written examination prior to the course and a self-confidence questionnaire, written examination and course evaluation when they finished the course. RESULTS From July 7, 2003 to March 8, 2005, 497 consenting learners completed the course. After course completion, learners demonstrated significant increases in their knowledge of terrorism response (t = -64.3, df = 496, p < 0.05) and their confidence in responding to terrorist events (t = -45.5, df = 496, p < 0.05). Learner feedback about the course was highly positive. CONCLUSIONS We successfully implemented a two-day course for professionals likely to respond to terrorist acts that included scenario-based performance training and assessment. Course participants increased their knowledge and were more confident in their ability to respond to acts of terrorism after participating in this course.


Toronto strives for normality after WHO lifts travel warning.


Why Your Rural Patients May Know More About Coronavirus Than You.


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

text: Increasing international trade in poultry meat presents an opportunity for the global dissemination of poultry disease. However, it would be very unfortunate if expanding world trade resulted in animal diseases being used as unjustified non-tariff trade barriers. For those avian diseases currently listed by the World Organisation for Animal Health, the current evidence suggests that only highly pathogenic avian influenza, Newcastle disease, and (for chicken meat) infectious bursal disease should be considered likely to be spread though trade in this commodity.


Infection control and prevention in home healthcare: prevention activities are the key to desired patient outcomes.


title: The 15th International Society of Aerosols in Medicine Congress. Perth, Australia, 14-18 March 2005.

text: This international meeting brought together approximately 250 delegates from the pharmaceutical industry, academia, hospitals and government agencies, to discuss the latest research and development on areas related to inhalation aerosols. Fundamental science and applied research encompassing both the biological and physicochemical aspects were presented. There was a wide range of topics covered, from immune modulation to pharmaceutical regulatory issues, including aerosol clearance; industry innovations; aerosols and in utero effects; technical advances in imaging; inhalation catastrophes; as well as recent advances and future directions in aerosol delivery systems. This biennial congress has provided an excellent forum for stimulating fruitful discussion of aerosols in medicine.


title: Comparative usability testing of conventional and single incision laparoscopic surgery devices.

text: OBJECTIVE The objective was to perform competitive usability testing to assess the user experience of conventional laparoscopic and laparoendoscopic single-site surgery (LESS) devices. BACKGROUND Recent advancements in single-incision instrumentation have created more interest in and usage of LESS. However, neither LESS nor its novel multichannel access devices have been thoroughly studied. METHOD Using a simulation test bed and standardized laparoscopic surgery task, the user experience of three commercially available LESS devices was compared to conventional laparoscopic ports based on time on task, errors, task success, and perceived ease of use. RESULTS There were no significant differences between devices for time on task, errors, or task success (p > .05). For all devices, there were significantly more recoverable than unrecoverable errors, and errors occurred more frequently during the second phase of the task when the dominant hand was more active (p < .0001). Conventional laparoscopy was rated as easier to use than were the LESS devices (p < .01). CONCLUSION Device performance of a basic laparoscopic task was similar in both conventional laparoscopy and LESS. Each of the LESS devices facilitated efficient and accurate aiming and grasping movements compared to conventional laparoscopy. Further investigation of human factors and ergonomics of LESS is needed to further develop, evaluate, and refine single-site surgery technologies to create a user experience equivalent to conventional laparoscopy. APPLICATION Competitive usability testing of medical devices yields objective performance data that can be used to inform purchase decisions and future device design improvements.


Thinking globally.


title: An account of the first hours of the Covid-19 epidemic at the Nephrology Unit in Lodi (Lombardy).

text: Marco Farina and colleagues give us their account of the first days of the Covid-19 epidemic in the Nephrology Unit of the Ospedale Maggiore in Lodi. From the news trickling through from Codogno on the 20th of February to the hospitalization, the following day, of the first dialytic patient with signs of pneumonia, who later tested positive to the virus. They tell us of how the hospital has been completely restructured in the wake of the epidemic, at remarkable speed and providing an example for others to follow, and the great sense self-sacrifice displayed by all medical personnel. After an overview of the clinical conditions of the 7 patients positive to the virus hospitalised in the following few days, they describe in some detail how symptomatic Covid+ patients are currently managed at the Ospedale Maggiore in Lodi.


title: Osteoid osteoma: CT-guided radiofrequency ablation.

text: Osteoid osteoma is a benign bone tumour; its main symptom is pain, which is sometimes resistant to analgesic or anti-inflammatory medication. The surgical treatment consists of en bloc excision or curettage of the lesion. Several alternative methods have been proposed, among which CT guided-percutaneous radiofrequency ablation. We report on ten patients who were diagnosed clinically and radiologically as presenting an osteoid osteoma and were treated with this technique, with more than two years follow-up. Results were uniformly excellent. The pain was relieved and the rate of post-operative complications was very low. We recommend percutaneous CT-guided radiofrequency ablation for the treatment of this lesion.


Functional analysis of the coronavirus MHV-JHM surface glycoproteins in vaccinia virus recombinants.


[Cognitive functions of HIV/AIDS positive individuals attending a comprehensive care program].


title: Cell-penetrating-peptide-based delivery of oligonucleotides: an overview.

text: Cationic CPPs (cell-penetrating peptides) have been used largely for intracellular delivery of low-molecular-mass drugs, biomolecules and particles. Most cationic CPPs bind to cell-associated glycosaminoglycans and are internalized by endocytosis, although the detailed mechanisms involved remain controversial. Sequestration and degradation in endocytic vesicles severely limits the efficiency of cytoplasmic and/or nuclear delivery of CPP-conjugated material. Re-routing the splicing machinery by using steric-block ON (oligonucleotide) analogues, such as PNAs (peptide nucleic acids) or PMOs (phosphorodiamidate morpholino oligomers), has consequently been inefficient when ONs are conjugated with standard CPPs such as Tat (transactivator of transcription), R(9) (nona-arginine), K(8) (octalysine) or penetratin in the absence of endosomolytic agents. New arginine-rich CPPs such as (R-Ahx-R)(4) (6-aminohexanoic acid-spaced oligo-arginine) or R(6) (hexa-arginine)-penetratin conjugated to PMO or PNA resulted in efficient splicing correction at non-cytotoxic doses in the absence of chloroquine. SAR (structure-activity relationship) analyses are underway to optimize these peptide delivery vectors and to understand their mechanisms of cellular internalization.


title: Cytotoxic T-cell-resistant variants arise at early times after infection in C57BL/6 but not in SCID mice infected with a neurotropic coronavirus.

text: Under certain conditions, C57BL/6 mice persistently infected with mouse hepatitis virus strain JHM (MHV-JHM) develop clinical disease and histological evidence of demyelination several weeks after inoculation with virus. In a previous report, we showed that mutations in the RNA encoding an immunodominant CD8 T-cell epitope within the surface glycoprotein (epitope S-510-518) were present in all persistently infected animals and that these mutations abrogated recognition by virus-specific cytotoxic T cells (CTLs) in direct ex vivo cytotoxicity assays. To obtain further evidence that these mutations were necessary for the development of clinical disease, the temporal course of their appearance was determined. Mutations in the epitope were identified by 10 to 12 days after inoculation, and in some mice, virus containing mutated epitope was the dominant species detected by 15 days. In addition, most mice that remain asymptomatic at 80 days after inoculation, a time after which clinical disease almost never develops, were infected with only wild-type virus. Finally, analysis of virus isolated from mice with severe combined immunodeficiency (SCID) revealed the presence only of wild-type epitope S-510-518. These results, by showing that mutations are not selected in SCID mice and occur at early times after inoculation in C57BL/6 mice, support the view that they result from immune pressure and contribute to virus persistence and demyelination in mice infected persistently with MHV-JHM.


title: Severe acute respiratory syndrome: radiographic and CT findings.

text: OBJECTIVE We review the radiographic and CT findings in the lungs of 12 patients with severe acute respiratory syndrome (SARS) in an effort to describe the most common radiologic findings for this disease. CONCLUSION The most common radiographic findings of SARS patients at presentation are unilateral or bilateral ground-glass opacities or focal unilateral or bilateral areas of consolidation. In hospitalized SARS patients, the abnormalities tend to progress to bilateral air-space consolidation. CT may reveal parenchymal disease in patients whose radiographs show normal results.


title: [cDNAs cloning of SARS-CoV PUMC2 viral genome].

text: OBJECTIVE To get the cDNA clones which cover the whole genome of SARS-CoV PUMC2 strain. METHODS Using the SARS-CoV PUMC2 strain genomic RNA as the template, the cDNA fragments were amplified by RT-PCR, the PCR products were further purified and ligated into the pGEM-T vector, and all the clones obtained were sequenced. RESULTS The cDNA clones which cover the whole genome of SARS-CoV PUMC2 strain were obtained. CONCLUSIONS These cDNAs can be provided for the function study of SARS-CoV proteins and the construction of full-length infectious cDNA clone of SARS-CoV.


title: Surface analysis of long-term hemodialysis catheters made of carbothane (poly(carbonate)urethane) before and after implantation in the patients' bodies.

text: PURPOSE The vascular cannulation is associated with a number of complications. The aim of this work was to study the composition and distribution of the film covering the surfaces of Mahurkar Maxid and Palindrome catheters, which were removed from the body of long-term hemodialysis patients. Moreover, the roughness and contact angle of the catheters were evaluated. METHODS Two brand new (as a reference) and thirty used catheters were the subject of the study. Their implantation period lasted from 4 months to a year and the reason for removal was the production of another vascular access or obstruction. Surfaces were analyzed by scanning electron microscope, atomic force microscope and goniometer. RESULTS The inner surfaces of the used catheters were covered with a film of various complexity which includes a plurality of protein, blood cell counts and the crystals. The closer to the distal part the film becomes more complex and multi-layered. Even the surfaces of brand new catheter were not completely smooth. The only significant difference between analyzed models was the presence of thrombus in the distal part of Mahurkar Maxid catheters, not in the Palindrome. CONCLUSIONS The distal part of the catheters is the place most exposed to obstruction and infection, which may be due to not reaching the anticoagulant agent into this part. Not only the occurrence of side holes affects the formation of thrombus, but also their quantity, geometry and distribution which effect on fluid mechanics. The surface of the catheters needs to improvement to minimize the occurrence of defects and cracks.


title: Plant-derived vaccines against diarrhoeal diseases.

text: Transgenic plant-derived vaccines offer a new strategy for the development of safe, inexpensive vaccines against diarrhoeal diseases. In animal and Phase I clinical studies, these vaccines have been safe and immunogenic without the need for a buffer or vehicle other than the plant cell. This review examines some early attempts to develop oral transgenic plant vaccines against enteric infections such as enterotoxigenic Escherichia coli infection, cholera and norovirus infection.


title: Childhood severe acute respiratory syndrome in Taiwan and how to differentiate it from childhood influenza infection.

text: OBJECTIVE To investigate clinical features and outcomes of children in Taiwan with laboratory-confirmed severe acute respiratory syndrome (SARS) vs those of children with influenza to differentiate the 2 diseases. DESIGN, SETTING, AND PARTICIPANTS Patients 20 years or younger with clinical, epidemiological, and laboratory evidence of SARS from March to July 2003 vs children with virus culture-confirmed influenza in a 1:1 age- and sex-matched control group. MAIN OUTCOME MEASURES Rates of symptoms, abnormal laboratory data, and outcomes of recovery, sequelae, or death. RESULTS The 15 SARS patients (9 girls and 6 boys) had a median age of 17 years (age range, 4-20 years). Nine patients (60%) were infected through household contact, 4 (27%) nosocomially, 1 (7%) through contact with a neighbor, and 1 (7%) after returning from Hong Kong. All 15 patients had fever, 3 (20%) had chills, and 11 (73%) had cough. Only 1 patient (7%) had sputum production; 1 (7%) had rhinorrhea. At presentation, 5 patients (33%) had leukopenia, 6 (40%) had lymphopenia, and 5 (33%) had monocytopenia. All children recovered without sequelae. Children with SARS had significantly lower incidences of rhinorrhea (odds ratio [OR], 0.01; 95% confidence interval [CI], 0.00-0.09), sputum production (OR, 0.10; 95% CI, 0.02-0.63), and sore throat (OR, 0.17; 95% CI, 0.03-0.85) than children with influenza. Both groups had similar incidences of leukopenia or lymphopenia, but SARS patients had a significantly higher incidence of monocytopenia (33% vs 0%, P = .04). CONCLUSIONS Childhood SARS is usually not fatal. The absence of rhinorrhea and presence of monocytopenia in SARS may distinguish it from influenza.


title: Laboratory-based surveillance for influenza: role of the Wisconsin State Laboratory of Hygiene.

text: Influenza poses a significant threat to public health worldwide. In the United States alone, mortality attributed to annual epidemics of influenza is estimated at 36,000 deaths per year. Influenza viruses also cause pandemics, during which the rates of illness and death can be expected to be much higher. In order to optimize prevention and control strategies for influenza, a variety of surveillance activities are carried out year-round internationally, nationally, and at the state level. This article summarizes relevant features of influenza and the surveillance activities carried out at each of these levels to monitor influenza activity. Particular emphasis is given to the state's laboratory-based surveillance network, developed and coordinated by the Wisconsin State Laboratory of Hygiene, that features strong and productive partnerships with private sector clinicians and laboratories and that provides a solid foundation for surveillance directed at other public health threats such as Severe Acute Respiratory Syndrome and bioterrorism.


title: Comparison of vaginal and abdominal hysterectomy:A prospective non-randomized trial.

text: OBJECTIVE To compare outcomes of vaginal and abdominal hysterectomy procedures in women with benign gynaecological diseases. METHODS This was a prospective study of outcomes of consecutive patients who underwent total vaginal hysterectomy (VH) or abdominal hysterectomy (AH) for benign gynaecological diseases. Patient characteristics before, during, and after the operations were reviewed. Patients were followed up for three months to evaluate postoperative complications. RESULTS This study included a total of 313 patients. 143 patients underwent AH and 170 patients underwent VH. Baseline characteristics were similar between the two groups. There were no intraoperative complications in either group. Operation time, intraoperative blood loss, first postoperative flatus time, time to out-of-bed activity, mean maximum postoperative body temperature, and duration of fever were all significantly shorter and less severe in the VH group compared with the AH group. In addition, vaginal length in the VH group was significantly shorter than in the AH group. CONCLUSIONS Vaginal hysterectomy has advantages over AH in the treatment of benign gynaecological diseases, providing greater efficacy and safety with minimal invasiveness.


title: Clinical characteristics and mechanism of liver damage in patients with severe acute respiratory syndrome.

text: BACKGROUND Abnormal liver function was found in patients with severe acute respiratory syndrome (SARS). This study was undertaken to explore the clinical characteristics and mechanisms of liver damage. METHODS The serial laboratory data of liver function test and hepatic histological examination from 168 patients with SARS were retrospectively analyzed. RESULTS The abnormalities of serum alanine aminotransferase (ALT) were 52.5%, 71.8%, 85.7% and 85.2%. The average levels of ALT of the patients were 56.07+/-51.57 U/L, 86.46+/-69.93 U/L, 106.69+/-102.50 U/L and 111.32+/-160.24 U/L, and the average levels of serum albumin were 37.25+/-5.37 g/L, 35.82+/-4.74 g/L, 34.49+/-5.04 g/L, and 34.26+/-4.70 g/L, at the day of admission, the first week, second week, and third week after hospitalization, respectively. Significant correlation was not shown among liver damage, blood oxygen saturation (SaO2), degree of fever, and immune functional disorder in this study. Hepatic histological examination of 4 patients demonstrated that non-specific inflammation existed in the liver. CONCLUSIONS Liver damage of patients with SARS usually occurs in the early stage of the disease with a high occurrence rate and a prolonged profile, which can be characterized by early, obvious decrease of albumin levels and slightly abnormal levels of ALT. The liver damage induced by SARS seems to be caused by SARS virus directly rather than by low SaO2 or high fever. Hepatotoxic drugs may play a role in increasing the severity of liver damage or prolonging the time of liver function recovery.


The pharmacist's role.


title: Taking a Toll on human disease: Toll-like receptor 4 agonists as vaccine adjuvants and monotherapeutic agents.

text: Toll-like receptor (TLR) agonists are being developed for use as vaccine adjuvants and as stand-alone immunomodulators because of their ability to stimulate innate and adaptive immune responses. Among the most thoroughly studied TLR agonists are the lipid A molecules that target the TLR4 complex. One promising candidate, monophosphoryl lipid A, which is a derivative of lipid A from Salmonella minnesota, has proven to be safe and effective as a vaccine adjuvant in > 120,000 human doses. A new class of synthetic lipid A mimetics, the aminoalkyl glucosaminide 4-phosphates (AGPs), have been engineered specifically to target human TLR4 and are showing promise as vaccine adjuvants and as monotherapeutic agents capable of eliciting nonspecific protection against a wide range of infectious pathogens. In this review, the authors provide an update of the preclinical and clinical experiences with the TLR4 agonists, MPL (Corixa Corporation) adjuvant and the AGPs.


title: Blood phosphorus and magnesium levels in 130 elite track and field athletes.

text: PURPOSE This study tested the clinical utility and relevance of serum phosphorus and magnesium as markers possibly useful to monitor training in athletes. METHODS Phosphorus and magnesium serum concentrations of 130 elite track and field athletes (65 males and 65 females, age range 20-30 years) from the National Athletics Sports Medicine Center database in Thessaloniki, Greece were measured. RESULTS Abnormal results were found in 61 (47%) athletes (32 men and 29 women). In male athletes, serum phosphate was higher than normal in 18% and decreased in 1.5%, whereas serum magnesium concentration was higher in 26%, and lower in 3%. Regarding female athletes, higher serum phosphate and magnesium levels were detected in 26% and 17% respectively, whereas decreased serum magnesium was found in 3%. The most common alterations were higher serum phosphate (29/61, 47%) and magnesium concentrations (28/61, 46%). Abnormalities of serum phosphorus and magnesium concentrations were detected in almost half of the athletes. Hyperphosphataemia and hypermagnesaemia were the most common abnormalities. CONCLUSION The reference intervals used for general population cannot be used for athletes. Given the lack of pathological manifestations, the physiopathological significance of these findings is uncertain. Further studies on the interpretation of reported ion concentrations in athletes should take in account the type of sport practiced and also the possible variations during the training and competition season.


title: The path of messenger RNA through the ribosome.

text: Using X-ray crystallography, we have directly observed the path of mRNA in the 70S ribosome in Fourier difference maps at 7 A resolution. About 30 nucleotides of the mRNA are wrapped in a groove that encircles the neck of the 30S subunit. The Shine-Dalgarno helix is bound in a large cleft between the head and the back of the platform. At the interface, only about eight nucleotides (-1 to +7), centered on the junction between the A and P codons, are exposed, and bond almost exclusively to 16S rRNA. The mRNA enters the ribosome around position +13 to +15, the location of downstream pseudoknots that stimulate -1 translational frame shifting.


title: Use of prediction markets to forecast infectious disease activity.

text: Prediction markets have accurately forecasted the outcomes of a wide range of future events, including sales of computer printers, elections, and the Federal Reserve's decisions about interest rates. We propose that prediction markets may be useful for tracking and forecasting emerging infectious diseases, such as severe acute respiratory syndrome and avian influenza, by aggregating expert opinion quickly, accurately, and inexpensively. Data from a pilot study in the state of Iowa suggest that these markets can accurately predict statewide seasonal influenza activity 2-4 weeks in advance by using clinical data volunteered from participating health care workers. Information revealed by prediction markets may help to inform treatment, prevention, and policy decisions. Also, these markets could help to refine existing surveillance systems.


title: Proportional assist ventilation versus pressure support ventilation in weaning ventilation: a pilot randomised controlled trial.

text: OBJECTIVE Proportional assist ventilation with load-adjustable gain factors (PAV+) is a mode of ventilation that provides assistance in proportion to patient effort. This may have physiological and clinical advantages when compared with pressure support ventilation (PSV). Our objective was to compare these two modes in patients being weaned from mechanical ventilation. DESIGN Prospective randomised controlled trial comparing PSV with PAV+. SETTING University-affiliated, tertiary referral intensive care unit (ICU). PARTICIPANTS Mechanically ventilated patients on a controlled mode of ventilation for at least 24 hours, who were anticipated to be spontaneously ventilated for at least 48 hours after randomisation. INTERVENTIONS Nil. MAIN OUTCOME MEASURES The primary outcome was time to successful liberation from the ventilator after the commencement of a spontaneous mode of ventilation. Secondary outcomes were requirement of rescue (mandatory) ventilation, requirement of sedative drugs, requirement for tracheostomy, re-intubation within 48 hours of extubation, ICU length of stay (LOS), hospital LOS, and ICU and hospital mortality. RESULTS 50 patients were randomised to either PSV (n = 25) or PAV+ (n = 25). There was no significant difference between the PAV+ and PSV groups in time to successful weaning (84.3 v 135.9 hours, respectively; P = 0.536). Four patients randomised to PAV+ were crossed over to PSV during weaning. There was no significant difference between groups for rescue ventilation, reintubation within 48 hours, tracheostomy, sedatives and analgesics prescribed, and ICU and hospital LOS. ICU mortality was higher in the PSV group (25% v 4 %; P = 0.002). CONCLUSIONS Both modes of ventilation were comparable in time to liberation from the ventilator.


title: [Virus inactivation by hydrogen peroxide].

text: The effect of H2O2 on adenovirus types 3 and 6, adenoassociated virus type 4, rhinoviruses 1A, 1B, and type 7, myxoviruses, influenza A and B, respiratory syncytial virus, strain Long, and coronavirus strain 229E was studied in vitro, using different H2O2 concentration and timec of exposure. H2O2 in a 3 percent concentration inactivated all the viruses under study within 1--30 min. Coronavirus and influenza viruses were found to be most sensitive. Reoviruses, adenoviruses and adenoassociated virus were relatively stable. H2O2 is a convenient means for virus inactivation.


title: How do people with knee osteoarthritis use osteoarthritis pain medications and does this change over time? Data from the Osteoarthritis Initiative.

text: INTRODUCTION The aim of this analysis was to describe comprehensively the cross-sectional and longitudinal patterns of analgesic and nutraceutical medication use for knee osteoarthritis (OA) in a contemporary US cohort and to investigate associated demographic and clinical factors. METHODS Baseline, 12, 24 and 36 month data were obtained retrospectively from the National Institutes of Health Osteoarthritis Initiative. Participants had symptomatic radiographic knee OA. Multiple binary logistic regression models identified characteristics independently associated with the use of analgesics or nutraceuticals. RESULTS We included 987 subjects (55.9% female, mean age 61.5 years, 71.0% white). At baseline, 68.2% reported frequent use of a conventional analgesic or nutraceutical for joint pain (for more than half of the previous month). Non-prescription non-steroidal anti-inflammatory drugs (NSAIDs) were the most frequently reported medications (26.8%), even in those more than 75-years old. Multiple conventional analgesics were used by 11.9%. Frequent analgesic use was more likely in women (odds ratio (OR) 1.8 (95% confidence interval (CI) 1.3 to 2.3)) and people with more pain (moderate 1.7 (1.2 to 2.4); severe 3.1 (2.1 to 4.7)); nutraceutical use was less likely in non-whites (0.4 (0.3 to 0.6)), those more than 74-years old (0.6 (0.3 to 0.9)) and those with comorbidities (0.6 (0.5 to 0.9)) and more likely in people with Kellgren-Lawrence (KL) grade 4 (2.2 (1.5 to 3.3)). Overall there was no change in the proportion of participants frequently using prescription or over the counter (OTC) analgesics at 36 months, although most people had changed medication type; of those using a traditional analgesic at baseline approximately one third were still using the same type at 36 months (ranging from 26.2% of baseline prescription NSAID users to 40.6% of baseline acetaminophen users). All participants reporting baseline analgesic use also reported 36 month analgesic use. Female participants (OR 95% CI 1.2 to 3.2, P = 0.009), those with high body mass index (1.2 to 4.8, P = 0.010) and those with moderate (1.6 to 2.6, P = 0.090) or severe (1.8 to 12.0, P = 0.002) baseline pain were more likely to use pain medication during the 36 month follow-up period; participants more than 75-years old were less likely (0.2 to 1.0, P = 0.053). CONCLUSIONS Most people with knee OA used pharmacological therapies frequently, and use appeared to be according to American College of Rheumatology recommendations. Change in medication type used was common. Persistent non-prescription NSAID use in older people is an area of concern.


title: Immunology taught by lung dendritic cells.

text: Dendritic cells (DCs) are leukocytes specialised in the uptake, processing, and presentation of antigen and fundamental in regulating both innate and adaptive immune functions. They are mainly localised at the interface between body surfaces and the environment, continuously scrutinising incoming antigen for the potential threat it may represent to the organism. In the respiratory tract, DCs constitute a tightly enmeshed network, with the most prominent populations localised in the epithelium of the conducting airways and lung parenchyma. Their unique localisation enables them to continuously assess inhaled antigen, either inducing tolerance to inoffensive substances, or initiating immunity against a potentially harmful pathogen. This immunological homeostasis requires stringent control mechanisms to protect the vital and fragile gaseous exchange barrier from unrestrained and damaging inflammation, or an exaggerated immune response to an innocuous allergen, such as in allergic asthma. During DC activation, there is upregulation of co-stimulatory molecules and maturation markers, enabling DC to activate naïve T cells. This activation is accompanied by chemokine and cytokine release that not only serves to amplify innate immune response, but also determines the type of effector T cell population generated. An increasing body of recent literature provides evidence that different DC subpopulations, such as myeloid DC (mDC) and plasmacytoid DC (pDC) in the lungs occupy a key position at the crossroads between tolerance and immunity. This review aims to provide the clinician and researcher with a summary of the latest insights into DC-mediated pulmonary immune regulation and its relevance for developing novel therapeutic strategies for various disease conditions such as infection, asthma, COPD, and fibrotic lung disease.


title: [Psychiatry in the time of pandemic : forging new ways without getting lost].

text: Coronavirus pandemic challenges mental health care providers in different ways. At an individual level, they have to adapt their clinical practice and consider specific issues of teleconsultations to address patient's needs. At a population level, dedicated interventions (e.g. hotlines, health professionals' support) have to strike a balance between « pathologizing » people's reactions and trivializing the psychological effects of a pandemic. Finally, psychiatric leaders must be able to advise decision-makers on mental-health policy and communication.


title: Development of reflective biosensor using fabrication of functionalized photonic nanocrystals.

text: Photonic crystals (PCs) are periodic dielectric structures that have a band-gap that forbids propagation of a certain range of wavelengths of light. This property enables control of light with remarkable facility by modification of the band-gaps and produce effects that are impossible with conventional optics. Using chemically functionalized PCs, where the chemical functional group consists of amine and carboxyl group, in conjunction with a biomolecular probe material, the detection of pathogens and viral disease is possible, indicated by the shift in wavelength signal. Moreover, this system using the bioinspired PCs allows specific target detection in biosensor chip fields through control of the PCs. In this study, we demonstrated that two bacterial pathogens (Fusobacterium necrophorum and Acinetobacter baumannii) causing sepsis were detected by DNA-probe hybridization and a severe acute respiratory syndrome coronavirus was detected by antigen-antibody interaction using the functional PCs. Optical readout with the integrated sensor detecting the signals from PCs, allows for low cost and robust readout of resonance peak shift. This biosensor system using the functional PCs on the photonic crystal-fabricated chip can efficiently and effectively detect various targets, and be easily prepared with high productivity and economic property.


title: A role for naturally occurring variation of the murine coronavirus spike protein in stabilizing association with the cellular receptor.

text: Murine hepatitis virus (MHV), a coronavirus, initiates infection by binding to its cellular receptor (MHVR) via spike (S) proteins projecting from the virion membrane. The structures of these S proteins vary considerably among MHV strains, and this variation is generally considered to be important in determining the strain-specific pathologies of MHV infection, perhaps by affecting the interaction between MHV and the MHVR. To address the relationships between S variation and receptor binding, assays capable of measuring interactions between MHV and MHVR were developed. The assays made use of a novel soluble form of the MHVR, sMHVR-Ig, which comprised the virus-binding immunoglobulin-like domain of MHVR fused to the Fc portion of human immunoglobulin G1. sMHVR-Ig was stably expressed as a disulfide-linked dimer in human 293 EBNA cells and was immobilized to Sepharose-protein G via the Fc domain. The resulting Sepharose beads were used to adsorb radiolabelled MHV particles. At 4 degrees C, the beads specifically adsorbed two prototype MHV strains, MHV JHM (strain 4) and a tissue culture-adapted mutant of MHV JHM, the JHMX strain. A shift to 37 degrees C resulted in elution of JHM but not JHMX. This in vitro observation of JHM (but not JHMX) elution from its receptor at 37 degrees C was paralleled by a corresponding 37 degrees C elution of receptor-associated JHM (but not JHMX) from tissue culture cells. The basis for this difference in maintenance of receptor association was correlated with a large deletion mutation present within the JHMX S protein, as sMHVR-Ig exhibited relatively thermostable binding to vaccinia virus-expressed S proteins containing the deletion. These results indicate that naturally occurring mutations in the coronavirus S protein affect the stability of the initial interaction with the host cell and thus contribute to the likelihood of successful infection by incoming virions. These changes in virus entry features may result in coronaviruses with novel pathogenic properties.


title: Mutual interference between genomic RNA replication and subgenomic mRNA transcription in brome mosaic virus.

text: Replication by many positive-strand RNA viruses includes genomic RNA amplification and subgenomic mRNA (sgRNA) transcription. For brome mosaic virus (BMV), both processes occur in virus-induced, membrane-associated compartments, require BMV replication factors 1a and 2a, and use negative-strand RNA3 as a template for genomic RNA3 and sgRNA syntheses. To begin elucidating their relations, we examined the interaction of RNA3 replication and sgRNA transcription in Saccharomyces cerevisiae expressing 1a and 2a, which support the full RNA3 replication cycle. Blocking sgRNA transcription stimulated RNA3 replication by up to 350%, implying that sgRNA transcription inhibits RNA3 replication. Such inhibition was independent of the sgRNA-encoded coat protein and operated in cis. We further found that sgRNA transcription inhibited RNA3 replication at a step or steps after negative-strand RNA3 synthesis, implying competition with positive-strand RNA3 synthesis for negative-strand RNA3 templates, viral replication factors, or common host components. Consistent with this, sgRNA transcription was stimulated by up to 400% when mutations inhibiting positive-strand RNA3 synthesis were introduced into the RNA3 5'-untranslated region. Thus, BMV subgenomic and genomic RNA syntheses mutually interfered with each other, apparently by competition for one or more common factors. In plant protoplasts replicating all three BMV genomic RNAs, mutations blocking sgRNA transcription often had lesser effects on RNA3 accumulation, possibly because RNA3 also competed with RNA1 and RNA2 replication templates and because any increase in RNA3 replication at the expense of RNA1 and RNA2 would be self-limited by decreased 1a and 2a expression from RNA1 and RNA2.


Intraoperative subarachnoid haemorrhage associated with a reduction in bispectral index during cardiopulmonary bypass.


title: Towards a new paradigm linking virus molecular evolution and pathogenesis: experimental design and phylodynamic inference.

text: Phylogenetic analysis has become a powerful tool for the investigation of evolution at a molecular level. During the last three decades, statistical phylogenetics has increasingly been applied to the study of microbial pathogens. The new field of phylodynamics was formally introduced in 2004 and encompasses the interaction between evolutionary and ecological processes that shape the spatiotemporal and phylogenetic patterns of infectious disease dynamics. This novel framework has significantly enhanced the study of measurable evolving pathogen populations, in particular RNA viruses and retroviruses. One of the major challenges in phylodynamic studies, however, is the generation of data in the form of dense coverage in sequence sampling coupled with high quality epidemiological and/or accurate clinical information. This review focuses specifically on experimental and data assembling strategies that are required to test multi-level phylodynamic hypotheses, ranging from intra-host viral evolution to population dynamics of infectious disease pandemics. Ultimately, bridging the gap between rational experimental design and phylodynamic inference will prove to be essential to take full advantage of this new exciting area of research.


Canine coronavirus infection in cats; a possible role in feline infectious peritonitis.


title: Substrate specificity profiling and identification of a new class of inhibitor for the major protease of the SARS coronavirus.

text: Severe acute respiratory syndrome (SARS) is an emerging infectious disease associated with a high rate of mortality. The SARS-associated coronavirus (SARS-CoV) has been identified as the etiological agent of the disease. Although public health procedures have been effective in combating the spread of SARS, concern remains about the possibility of a recurrence. Various approaches are being pursued for the development of efficacious therapeutics. One promising approach is to develop small molecule inhibitors of the essential major polyprotein processing protease 3Clpro. Here we report a complete description of the tetrapeptide substrate specificity of 3Clpro using fully degenerate peptide libraries consisting of all 160,000 possible naturally occurring tetrapeptides. The substrate specificity data show the expected P1-Gln P2-Leu specificity and elucidate a novel preference for P1-His containing substrates equal to the expected preference for P1-Gln. These data were then used to develop optimal substrates for a high-throughput screen of a 2000 compound small-molecule inhibitor library consisting of known cysteine protease inhibitor scaffolds. We also report the 1.8 A X-ray crystal structure of 3Clpro bound to an irreversible inhibitor. This inhibitor, an alpha,beta-epoxyketone, inhibits 3Clpro with a k3/Ki of 0.002 microM(-1) s(-1) in a mode consistent with the substrate specificity data. Finally, we report the successful rational improvement of this scaffold with second generation inhibitors. These data provide the foundation for a rational small-molecule inhibitor design effort based upon the inhibitor scaffold identified, the crystal structure of the complex, and a more complete understanding of P1-P4 substrate specificity.


title: Nontraumatic osteonecrosis of the femoral head: ten years later.

text: The etiology of osteonecrosis of the hip may have a genetic basis. The interaction between certain risk factors and a genetic predisposition may determine whether this disease will develop in a particular individual. The rationale for use of joint-sparing procedures in the treatment of this disease is based on radiographic measurements and findings with other imaging modalities. Early diagnosis and intervention prior to collapse of the femoral head is key to a successful outcome of joint-preserving procedures. The results of joint-preserving procedures are less satisfactory than the results of total hip arthroplasty for femoral heads that have already collapsed. New pharmacological measures as well as the use of growth and differentiation factors for the prevention and treatment of this disease may eventually alter our treatment approach, but it is necessary to await results of clinical research with long-term follow-up of these patients.


title: Outcome of esophagojejunostomy during totally laparoscopic total gastrectomy: a single-center retrospective study.

text: AIM The present study aimed to clarify the safety and feasibility of esophagojejunostomy during totally laparoscopic total gastrectomy (TLTG). PATIENTS AND METHODS In 45 consecutive patients who underwent TLTG for gastric cancer, esophagojejunostomy was performed with a functional end-to-end anastomosis (FEEA) using a linear stapler in 24 patients or with a double stapling technique (DST) using a trans-orally inserted anvil (OrVil™) in 21 patients. RESULTS The DST was more likely to be chosen in patients with tumors located in the upper stomach. In the FEEA group, both the mean length of the operation and the mean postoperative hospital stay were significantly shorter compared to those in the DST group. Two patients in the FEEA group and four patients in the DST group developed postoperative complications but there were no postoperative deaths in either group. CONCLUSION Both FEEA and DST in esophagojejunostomy during TLTG are safe and feasible.


title: Diagnosis of noncultivatable gastroenteritis viruses, the human caliciviruses.

text: Gastroenteritis is one of the most common illnesses of humans, and many different viruses have been causally associated with this disease. Of those enteric viruses that have been established as etiologic agents of gastroenteritis, only the human caliciviruses cannot be cultivated in vitro. The cloning of Norwalk virus and subsequently of other human caliciviruses has led to the development of several new diagnostic assays. Antigen detection enzyme immunoassays (EIAs) using polyclonal hyperimmune animal sera and antibody detection EIAs using recombinant virus-like particles have supplanted the use of human-derived reagents, but the use of these assays has been restricted to research laboratories. Reverse transcription-PCR assays for the detection of human caliciviruses are more widely available, and these assays have been used to identify virus in clinical specimens as well as in food, water, and other environmental samples. The application of these newer assays has significantly increased the recognition of the importance of human caliciviruses as causes of sporadic and outbreak-associated gastroenteritis.


title: [A novel one-tube multiplex RT-PCR assay for simultaneous detection of six human coronaviruses].

text: OBJECTIVE To develop an one-tube multiplex RT-PCR assay for simultaneous detection of six human coronaviruses (HCoVs). METHODS Genbank sequences of six human coronaviruses, including HCoV-NL63, HCoV-229E, SARS-CoV, HCoV-OC43, MERS-CoV, and HCoV-HKU1, were included as reference sequences. Primers were designed based on multiple alignment of reference sequences, targeting the conserved regions of each species of HcoV. Virus strains and nucleic acid preserved in our lab were used as template in developing this automatic-electrophoresis-based one-tube multiplex RT-PCR assay. Detection limits and reproducibility were also evaluated with these templates. Samples with infection of other respiratory viruses preserved in our lab were used to evaluate specificity of this assay. Finally, we tested this assay with 140 clinical samples that were validated by real-time PCR in parallel. RESULTS This automatic-electrophoresis-based multiplex RT-PCR assay was able to detect six human coronaviruses simultaneously. All positive samples in this study were detected with at least one specific fragment of anticipated length (195, 304, 332, 378, 415, 442 bp) . No fragment was detected in negative controls. Detection limits of 1.0×10(1-1.0)×10(2) copies/µl were achieved in tests of single virus. No cross reaction was observed with other respiratory viruses. This multiplex RT-PCR assay showed the same sensitivity and specificity to that of individual real-time RT-PCR validated with clinical samples. Both methods detected 28 positive samples (20%) . CONCLUSIONS Six HCoVs can be detected in one tube by this novel multiplex RT-PCR assay with high sensitivity and specificity.


title: Clinical associations of Aeromonas spp. in fecal specimens from children.

text: Human infections caused by Aeromonas spp. are being reported with increased frequency. We examined 1012 unselected stool specimens from 799 children for various pathogens, including Aeromonas spp., to evaluate the clinical associations of Aeromonas spp. in the stools of children in the East End of London. A total of 33 children had positive cultures for Aeromonas spp. These were identified as: Aeromonas hydrophila (16), A. sobria (9), A. caviae (8). Aeromonas spp. were identified by being oxidase positive in character, a property that distinguishes Aeromonas spp. from Enterobacteriaceae. Associated pathogens included Salmonella typhimurium (1), Campylobacter spp. (3), Enteropathogenic Escherichia coli (2), and Giardia lamblia (1). Electron microscopic examination of the stool of 13 children revealed viral particles in two, rotavirus (1), and coronavirus-like particles (1). The associated clinical features in the 25 children with only Aeromonas excretion were diarrhea (21), vomiting (10), abdominal pain (9), fever (4), and mild dehydration (2). Five patients had macroscopic blood loss in the stools. The illness was generally mild and lasted only a short time.


What does the RAND Health Insurance Experiment tell us about the impact of patient cost sharing on health outcomes?


title: Detection of infectious bronchitis virus with the use of real-time quantitative reverse transcriptase-PCR and correlation with virus detection in embryonated eggs.

text: Real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assays have been used to detect the presence of challenge virus when the efficacy of infectious bronchitis virus (IBV) vaccine against field viruses is being experimentally evaluated. However, federal guidelines for licensing IBV vaccines indicate that challenge-virus detection following vaccination is to be conducted in embryonated eggs. In this study, we examined qRT-PCR data with the use of universal and type-specific primers and probe sets for IBV detection and compared those data with challenge-virus detection in embryonated eggs to determine if the two methods of evaluating vaccine efficacy are comparable. In addition, we tested the qRT-PCR assays on thermocyclers from two different manufacturers. We found the universal IBV primers and probe set to be comparable to challenge-virus detection in embryonated eggs. However, for some IBV types (Mass41 and Conn on the SmartCycler II and Ark, Mass41, Conn, and GA98 on the ABI 7500) the qRT-PCR assay was more sensitive than virus detection in embryonated eggs. This may simply be due to the universal IBV qRT-PCR assay being more sensitive than virus detection in eggs or to the assay detecting nucleic acid from nonviable virus. This finding is important and needs to be considered when evaluating challenge-virus detection for vaccination and challenge studies, because qRT-PCR could potentially identify positive birds that would otherwise be negative by virus detection in embryonated eggs; thus it could lead to a more stringent measure of vaccine efficacy. We also found that the IBV type-specific primers and probe sets designed in this study were in general less sensitive than the universal IBV primers and probe set. Only the Ark-DPI-spedcific assay on the SmartCycler II and the Ark-DPI-, Mass41-, and DE072/GA98- (for detection of GA98 virus only) specific assays on the ABI 7500 were comparable in sensitivity to virus detection in eggs. We found that a number of variables, including the virus type examined, primers and probe efficiency and stability, and assay conditions, including thermocycler platform, can affect the data obtained from qRT-PCR assays. These results indicate that qRT-PCR assays can be used to detect IBV challenge virus, but each assay, including the assay conditions and thermocycler, should be individually evaluated if those data are expected to be comparable to virus detection in embryonated eggs.


title: Rapid awareness and transmission of severe acute respiratory syndrome in Hanoi French Hospital, Vietnam.

text: A case-control study was conducted to examine the relationship between severe acute respiratory syndrome (SARS) and the time-dependent precautionary behaviors taken during an outbreak of SARS in Hanoi French Hospital (HFH), Vietnam. Masks (odds ratio [OR] = 0.3; 95% confidence interval [CI]: 0.1, 0.7) and gowns (OR = 0.2; 95% CI: 0.0, 0.8) appeared to prevent SARS transmission. The proportion of doctors and nurses who undertook each measure significantly improved (chi(2) = 9.8551, P = 0.043) after the onset of secondary cases. The impact of individual behaviors on an outbreak was investigated through mathematical approaches. The reproduction number decreased from 4.1 to 0.7 after notification. The basic reproduction number was estimated, and the use of masks alone was shown to be insufficient in containing an epidemic. Intuitive results obtained by means of stochastic individual-based simulations showed that rapid improvements in behavior and isolation would increase the probability of extinction.