title: Preliminary studies of primary ostrich fibroblasts for the isolation of ratite viruses.

text: An ostrich egg at 21 days of development was used to propagate primary embryo cell cultures. Primary cultures of skeletal muscle cells (for fibroblasts) were prepared by routine typsinization techniques. The ostrich embryo fibroblasts were tested for their ability to propagate stock avian viruses of infectious bronchitis virus, paramyxiovirus-1 (PMV-1), PMV-2, PMV-3, infectious bursal disease virus, quail bronchitis virus, avian reovirus, turkey coronavirus, and two ostrich-originating specimens (one of which was a possible coronavirus identified by electron microscopy). Cytopathic effects were seen by light microscopy in cell cultures inoculated with PMV-1, turkey coronavirus, and the two ostrich specimens. Hemaglutinating titers of 4 or more were determined for PMV-1, turkey coronavrius, and the two ostrich specimens after inoculation onto monolayers of ostrich embryo fibroblasts. Hemagglutination-inhibition tests confirmed the identification of PMV-1 when homologous antisera were used as the specific inhibitor. Bovine coronavirus antisera inhibited the hemagglutination of one of the cultured ostrich specimens.


title: [Clinical and chest X-ray characteristics of 5 cases with severe acute respiratory syndrome in children in Shenzhen area].

text: OBJECTIVE To explore clinical and chest X-ray features of SARS in children to facilitate correct diagnosis. METHODS Clinical manifestations and chest X-ray findings in five children suffering from SARS admitted for treatment in the hospital between February and May, 2003 in Shenzhen area were analyzed. The diagnosis was confirmed by epidemiological, clinical, laboratory and radiological examinations. Among the 5 cases, 1 was a boy and the others were girls at the age of 4 to 13 years. RESULTS Of the 5 SARS children, 3 presented a history of close contact with SARS patients. Fever was the initiative symptom, 4 had a body temperature of over 38 degrees C with the highest being 40 degrees C; fever sustained from 4 to 7 days with an average of 5.6 days. All the 5 cases developed nonproductive cough; on auscultation, both moist and dry rales could be heard in 3 out of the 5 cases. Mean total white count of peripheral blood was (2.96 - 6.9) x 10(9)/L, and was < 5.0 x 10(9)/L in 4 cases. SARS associated coronavirus specific RNA fragment was found positive by RT-PCR in 1 case; 1 case was positive for both IgM and IgG antibodies to the virus; 1 case was positive for only IgM antibody and another 2 cases were positive for only IgG antibody. IgG and IgM antibodies to Mycoplasma pneumoniae and Chlamydia pneumoniae as well as blood culture for bacteria were all negative. Findings on chest X-ray examination: 4 cases showed presence of patchy or macular opacities with cord-like shadows in unilateral lung plates while 1 case each showed ground-glass-like opacity and migratory changes; 1 case showed interstitial changes in the lungs in the form of irregular reticular lattice and cord-like shadows. Two cases received CT scanning and macular-patchy or spotty shadows were seen all over the lung. The shortest time for absorption of foci in the lungs was 7 days while the longest was 33 days with a mean of 15 +/- 6 days. None of the cases had any signs of fibrosis in the lungs. All the 5 cases were completely cured and discharged 7 to 40 days (mean 18 +/- 11 days) after admission. CONCLUSION Compared with adult cases with SARS, children with SARS had milder symptoms and signs. Presence of unilateral patchy shadow in lungs represented the main chest X-ray findings.


title: Detection and differentiation of North American and European genotypes of porcine reproductive and respiratory syndrome virus in formalin-fixed, paraffin-embedded tissues by multiplex reverse transcription-nested polymerase chain reaction.

text: North American and European genotypes of porcine reproductive and respiratory syndrome virus (PRRSV) were distinguished in formalin-fixed, paraffin-embedded tissues from PRRSV-infected pigs by multiplex reverse transcription-nested polymerase chain reaction (RT-nPCR). A method based on xylene deparaffinization followed by proteinase K digestion yielded RNA of quality for reliable and consistent RT-nPCR analyses. The PRRSV nucleic acid was detected in lung, mediastinal lymph node, tonsil, and liver samples from pigs inoculated with North American strain virus, European strain virus, or both North American and European strains. All 30 archival formalin-fixed, paraffin-embedded lung tissues from pigs naturally infected with PRRSV were positive for PRRSV by RT-nPCR amplication and gave a pattern that corresponded to the North American genotype. Multiplex RT-nPCR could be applied successfully to formalin-fixed, paraffin-embedded tissues for the detection and differentiation between North American and European genotypes of PRRSV.


title: Type I interferon as a link between innate and adaptive immunity through dendritic cell stimulation.

text: Type I interferon (IFN-alpha/beta) is expressed rapidly after infection and plays a key role in innate defense against pathogens. Recent studies have shown that a connection exists between IFN-alpha/beta and antigen-presenting dendritic cells (DCs) at two levels. Firstly, a specific DC precursor, the plasmacytoid pre-DC (p-preDC), was identified as a cell type able to secrete very high amounts of IFN-alpha/beta following stimulation with infectious agents. Secondly, IFN-alpha/beta has been shown to act as a differentiation/maturation factor for DCs. These findings will be discussed in association with evidence indicating that IFN-alpha/beta can enhance and modulate immune responses in vivo. Taken together, the available data suggest that IFN-alpha/beta serves as a link between the innate response to infection and the adaptive immune response.


title: Viral interference with B7-1 costimulation: a new role for murine cytomegalovirus fc receptor-1.

text: Murine CMV (MCMV), a beta-herpesvirus, infects dendritic cells (DC) and impairs their function. The underlying events are poorly described. In this study, we identify MCMV m138 as the viral gene responsible for promoting the rapid disappearance of the costimulatory molecule B7-1 (CD80) from the cell surface of DC. This was unexpected, as m138 was previously identified as fcr-1, a putative virus-encoded FcR. m138 impaired the ability of DC to activate CD8+ T cells. Biochemical analysis and immunocytochemistry showed that m138 targets B7-1 in the secretory pathway and reroutes it to lysosomal associated membrane glycoprotein-1+ compartments. These results show a novel function for m138 in MCMV infection and identify the first viral protein to target B7-1.


title: Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens.

text: The emergence of a novel coronavirus (CoV) as the cause of severe acute respiratory syndrome (SARS) catalyzed the development of rapid diagnostic tests. Stool samples have been shown to be appropriate for diagnostic testing for SARS CoV, although it has been recognized to be a heterogeneous and difficult sample that contains amplification inhibitors. Limited information on the efficiency of extraction methods for the purification and concentration of SARS CoV RNA from stool samples is available. Our study objectives were to determine the optimal extraction method for SARS CoV RNA detection and to examine the effect of increased specimen volume for the detection of SARS CoV RNA in stool specimens. We conducted a multicenter evaluation of four automated and four manual extraction methods using dilutions of viral lysate in replicate mock stool samples, followed by quantitation of SARS CoV RNA using real-time reverse transcriptase PCR. The sensitivities of the manual methods ranged from 50% to 100%, with the Cortex Biochem Magazorb method, a magnetic bead isolation method, allowing detection of all 12 positive samples. The sensitivities of the automated methods ranged from 75% to 100%. The bioMérieux NucliSens automated extractor and miniMag extraction methods each had a sensitivity of 100%. Examination of the copy numbers detected and the generation of 10-fold dilutions of the extracted material indicated that a number of extraction methods retained inhibitory substances that prevented optimal amplification. Increasing the volume of sample input did improve detection. This information could be useful for the extraction of other RNA viruses from stool samples and demonstrates the need to evaluate extraction methods for different specimen types.


title: Evaluation of WHO criteria for identifying patients with severe acute respiratory syndrome out of hospital: prospective observational study.

text: OBJECTIVES To determine the clinical and radiological features of severe acute respiratory syndrome (SARS) and to evaluate the accuracy of the World Health Organization's guidelines on defining cases of SARS. DESIGN Prospective observational study. SETTING A newly set up SARS screening clinic in the emergency department of a university hospital in Hong Kong's New Territories. PARTICIPANTS 556 hospital staff, patients, and relatives who attended the screening clinic and who had had contact with someone with SARS. MAIN OUTCOME MEASURE Number of confirmed cases of SARS. RESULTS Of the 556 people, 141 were admitted to hospital, and 97 had confirmed SARS. Fever, chills, malaise, myalgia, rigor, loss of appetite, vomiting, diarrhoea, and neck pain but not respiratory tract symptoms were significantly more common among the 97 patients than among the other patients. The overall accuracy of the WHO guidelines for identifying suspected SARS was 83% and their negative predictive value was 86% (95% confidence interval 83% to 89%). They had a sensitivity of 26% (17% to 36%) and a specificity of 96% (93% to 97%). CONCLUSIONS Current WHO guidelines for diagnosing suspected SARS may not be sufficiently sensitive in assessing patients before admission to hospital. Daily follow up, evaluation of non-respiratory, systemic symptoms, and chest radiography would be better screening tools.


title: Concentration and detection of SARS coronavirus in sewage from Xiao Tang Shan Hospital and the 309th Hospital of the Chinese People's Liberation Army.

text: A worldwide outbreak of severe acute respiratory syndrome (SARS) had been reported. Over 8439 SARS cases and 812 SARS-related deaths were reported to the World Health Organization from 32 countries around the world up to 5 July 2003. The mechanism of transmission of SARS-CoV has been limited only to close contacts with patients. Attention was focused on possible transmission by the sewage system because laboratory studies showed that patients excreted coronavirus RNA in their stools in Amoy Gardens in Hong Kong. To explore whether the stool of SARS patients or the sewage containing the stool of patients would transmit SARS-CoV or not, we used a style of electropositive filter media particle to concentrate the SARS-CoV from the sewage of two hospitals receiving SARS patients in Beijing, as well as cell culture, semi-nested RT-PCR and sequencing of genes to detect and identify the viruses from sewage. There was no live SARS-CoV detected in the sewage in these assays. The nucleic acid of SARS-CoV was found in the sewage before disinfection from both hospitals by PCR. After disinfection, SARS-CoV RNA could be detected from some samples from the 309th Hospital of the Chinese People's Liberation Army, but not from Xiao Tang Shan Hospital after disinfection. In this study, we found that the virus can survive for 14 days in sewage at 4 degrees C, 2 days at 20 degrees C, and its RNA can be detected for 8 days though the virus had been inactivated. In conclusion, this study demonstrates that the RNA of SARS-CoV could be detected from the concentrates of sewage of both hospitals receiving SARS patients before disinfection and occasionally after disinfection though there was no live SARS-CoV; thus much attention should be paid to the treatment of stools of patients and the sewage of hospitals receiving SARS patients.


title: First description of Mycobacterium heckeshornense infection in a feline immunodeficiency virus-positive cat.

text: A 13-year-old cat was presented to the veterinary clinic with poor condition, vomiting and a reduced appetite. A painful abdomen was diagnosed because of tension and defence reactions on palpation. Diagnostic laparotomy showed a thickening of the colon and caecal intestinal wall. Histopathological investigation of intestinal biopsies revealed focal severe granulomatous inflammation with numerous acid-fast bacilli in the tela submucosa. The complete blood count test showed a severe lymphopenia and anaemia, and the cat tested positive for feline immunodeficiency virus (FIV) antibodies by enzyme-linked immunosorbent assay. The cat was euthanased and necropsied. Multifocal granulomatous nodules were present in the intestines, liver and kidneys. The gastric lymph node was markedly enlarged and showed a caseous cut surface. Histopathology revealed a systemic mycobacteriosis affecting intestine, lymph nodes, liver and kidneys. The mycobacterial strain was cultured and determined by its unique 16S rRNA gene sequence as Mycobacterium heckeshornense. This is the first reported case of M heckeshornense in a cat. It was suspected that the disseminated mycobacteriosis was supported by the FIV infection.


title: Safety and Effectiveness of Enoxaparin as Venous Thromboembolism Prophylaxis after Gastric Cancer Surgery in Japanese Patients.

text: Routine prophylaxis for venous thromboembolism (VTE) has been recommended after surgery not only in the West but also in Asia recently. The primary objective of this study was to investigate the safety and effectiveness of enoxaparin as a prophylaxis in patients undergoing distal, proximal, or total gastrectomy (TG) for gastric cancers. A total of 565 patients who underwent gastrectomy for gastric cancer were reviewed retrospectively. About 256 patients received postoperative prophylaxis with enoxaparin (2000 international unit twice daily for at least six days) and compression stockings; these patients were assigned to the enoxaparin group. About 257 patients comprised a historical control group, who used only compression stockings as a thromboprophylaxis. All patients underwent the same rehabilitation programs during the perioperative period. None of the patients developed symptomatic venous thromboembolism in either the enoxaparin group or the control group. The complication rate of bleeding was not significantly different between the two groups. Only one patient who used three antiplatelet agents concomitantly with enoxaparin required reoperation for anastomotic site bleeding. The usage of enoxaparin for venous thromboembolism prophylaxis is safe for Japanese patients after gastrectomy. But, cautious application is still needed especially when used concomitantly with other antiplatelet agents.


[Study on the stability of SARS coronavirus].


title: Molecular bases of tropism in the PUR46 cluster of transmissible gastroenteritis coronaviruses.

text: Transmissible gastroenteritis coronavirus (TGEV) infects both, the enteric and the respiratory tract of swine. S protein, that is recognized by the cellular receptor, has been proposed that plays an essential role in controlling the dominant tropism. The genetic relationship of S gene from different enteric strains and non-enteropathogenic porcine respiratory coronaviruses (PRCVs) was determined. A correlation between tropism and the genetic structure of the S gene was established. PRCVs, derived from enteric isolates have a large deletion at the N-terminus of the S protein. Interestingly, two respiratory isolates, attenuated Purdue type virus (PTV-ATT) and Toyama (TOY56) have a full-length S gene. PTV-ATT has two specific amino acid differences with the S protein of the enteric viruses. One is located around position 219, within the deleted area, suggesting that alterations around this amino acid may result in the loss of enteric tropism. To study the role of different genes in tropism, a cluster of viruses closely related to PUR46 strain was analyzed. All of them have been originated by accumulating point mutations from a common, virulent isolate which infected the enteric tract. During their evolution these viruses have lost, virulence first, and then, enteric tropism. Sequencing analysis proved that enteric tropism could be lost without changes in ORFs 3a, 3b, 4, 6, and 7, and in 3'-end untranslated regions (3'-UTR). To study the role of the S protein in tropism recombinants were obtained between an enteric and a respiratory virus of this cluster. Analysis of the recombinants supported the hypothesis on the role in tropism of S protein domain around position 219.


title: Small bowel occlusion after trans-abdominal preperitoneal hernia approach caused by barbed suture: case report and review of literature.

text: BACKGROUND Groin hernioplasty is the most performed intervention in the adults worldwide. Small bowel occlusion after hernioplasty with anterior approach is an unusual complication because the peritoneum is not opened during this procedure. However during TAPP the closure of the peritoneal flap is mandatory. In literature some cases of small bowel occlusion related to the barbed suture for the closure the peritoneum are reported. METHODS Here we describe a case of a 64-year old male with small bowel obstruction after TAPP caused by the barbed suture used for peritoneal closure. RESULTS Intrabdominal use of self-anchoring suture is controversial. Some studies reported good results by using this device, while others from gynecologists describe bowel occlusion and volvulus caused by barbed suture. CONCLUSIONS Self-anchoring device is innovative and reduces operation time. It is most important to know the correct use of this device to reduce some possible troubles.


title: How genomics has affected the concept of microbiology.

text: Genomics influences multiple areas of microbiology, and thus affects key microbiological concepts. Recent reports that describe the large genome and unusual coding capacity of mimivirus, the minimized fungal genomes that contain elements of bacterial metabolism, and the 'signature' eukaryotic proteins in bacteria are introducing grey shades into the black-and-white distinctions between microbial domains. The concept of the 'universal' minimal genome is being challenged, and the ability of minimal genomes to support cellular complexity is under investigation. There have been intriguing insights into microbe-microbe relationships, for example conflict mediation in competing bacteriophages that rapidly evolve survival mechanisms when cooperation is experimentally enforced. Genomics has given birth to metagenomics, but has also stimulated the development of improved cultivation techniques. Lastly, the taxonomic potential of genomics is emerging, as studies of multiple strains allow us to revise and refine the bacterial species concept as well as the idea of a static genome.


title: Exercise and chronic kidney disease: current recommendations.

text: Patients with chronic kidney disease (CKD) are inactive and have reduced physical functioning and performance. Aerobic exercise interventions have been shown to increase maximal oxygen consumption in selected patients. In addition, preliminary evidence, although mixed, suggests that aerobic exercise training can improve blood pressure control, lipid profiles and mental health in this population. A few larger studies are now available showing that aerobic training can also improve physical functioning and performance. The impact on survival or hospitalisation has not been determined. Resistance exercise training, although less studied, appears to increase muscle strength and size and may also improve functioning. There have been several reports of successful combined exercise interventions, but the designs have not allowed evaluation of the relative benefits of aerobic and resistance training on physical functioning. Despite the evidence that exercise is safe and beneficial in patients with CKD, dialysis patients remain inactive, and exercise assessment, counselling and training is not widely offered to patients with CKD. Studies of the barriers to patient participation in exercise and to provider assessment and recommendations are needed so that more widely generalisable interventions can be developed. However, in the interim, patients should be encouraged to participate in moderate physical activity to meet the US Surgeon General's recommendations. Patients who are weak can benefit from strength-training interventions. Resistance and aerobic exercise programmes should be initiated at relatively low intensity in patients with CKD and progressed as slowly as tolerated in order to avoid injury and discontinuation of exercise. For patients on haemodialysis, incorporation of exercise into the dialysis session may increase patient participation and tolerance of exercise.


title: Defining a reference range for vital signs in healthy term pregnant women undergoing caesarean section.

text: Early warning systems (EWS), used to identify deteriorating hospitalised patients, are based on measurement of vital signs. When the patients are pregnant, most EWS still use non-pregnant reference ranges of vital signs to determine trigger thresholds. There are no published reference ranges for all vital signs in pregnancy. We aimed to define vital signs reference ranges for term pregnancy in the preoperative period, and to determine the appropriateness of EWS trigger criteria in pregnancy. We conducted a one-year retrospective study in a tertiary referral obstetric hospital. The study sample was healthy term women undergoing planned caesarean section (CS). Systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), oxygen saturation (SpO2) and temperature were all measured automatically and data was extracted from the medical record. Two hundred and fifty-eight women met inclusion criteria. Results were (mean ± SD [standard deviation]) SBP 118 ± 11.2 mmHg, DBP 75 ± 10.3 mmHg, HR 84 ± 10.2 /minute, respiratory rate 18 ± 1.5 /minute, SpO2 99% ± 1.0% and temperature 36.4°C ± 0.43°C. The reference ranges (mean ± 2SD) determined were SBP 96-140 mmHg, DBP 54-96 mmHg, HR 64-104/minute, RR 15-21 /minute, SpO2 97%-100% and temperature 35.5°C-37.3°C. This study defined a reference range for vital signs in healthy term pregnant women undergoing CS. Study findings suggest that currently used criteria for EWS triggers, based on non-pregnant values, may be too extreme for timely detection of deteriorating pregnant patients. Further research examining the modified HR triggers of ≤50 and ≥110 /minute in pregnant women and their relationship to clinical outcomes is required.


title: A prospective etiologic study of community-acquired pneumonia in Taiwan.

text: BACKGROUND AND PURPOSE The treatment of community-acquired pneumonia (CAP) is complicated by the growing threat of antimicrobial resistance and the tendency to rely on empirical therapy. This study investigated the etiologic agents of adult CAP in Taiwan and the susceptibility of Streptococcus pneumoniae isolates from these patients. METHODS A collaborative group was established in the emergency department to conduct a prospective study of the etiology of adult CAP. The etiologic agent was determined by a combination of microscopic, culture, serologic and antigen detection methods. Pneumococcal susceptibility testing was performed to determine the extent of penicillin resistance. RESULTS A total of 100 consecutive cases of mild to moderate adult CAP prior to the severe acute respiratory syndrome epidemic were enrolled. The etiologic agent was determined in 72% of cases. The 5 most common causative pathogens were S. pneumoniae (26%), Mycoplasma pneumoniae (20%), Chlamydia pneumoniae (13%), Haemophilus influenzae (9%), and Klebsiella pneumoniae (5%). Atypical pathogens accounted for 40% of CAP. Bacteremic pneumonia was diagnosed in 6.2% of cases. Co-infections with 2 or more pathogens were found in 16% of the cases. Among the 20 isolates of S. pneumoniae, 85% (17/20) were susceptible to penicillin, 3 (15%) were intermediate, and none were resistant to penicillin. CONCLUSION S. pneumoniae, M. pneumoniae and C. pneumoniae were the 3 leading causes of mild to moderate CAP in Taiwan. This study indicates that penicillin-resistant S. pneumoniae play a very limited role in this condition in adults.


title: Concomitant sublay mesh repair of umbilical hernia and abdominoplasty.

text: Concomitant mesh repair of large umbilical hernias and abdominoplasty pose a serious risk of devascularizing the umbilical stalk. A technique of placing mesh in a sublay manner, deep to the fascial defect, for an umbilical herniorrhaphy to avoid damage to the deep umbilical perforators during an abdominoplasty is described.


title: Differences in sources of information used by the population between the affected area and the general population during the first phase of a bird flu outbreak.

text: INTRODUCTION In March 2006, a few cases of bird flu were discovered in approximately 10 rural settlements in Israel. As a result, approximately one million birds were destroyed within a three kilometer radius of the settlements. The Israeli population was instructed to take preventive measures against the spread of the infection. OBJECTIVES The objective of this study was to compare the frequency of use of different sources of information by the population in the affected area with the general population during the first phase of a bird flu outbreak in Israel. METHODS A telephone survey among two randomly selected, representative samples of adults was conducted. One sample involved 500 adult Israeli residents; the other sample involved 103 adult residents from the affected area during the first phase of the outbreak. The use of different sources of information by the population concerning the disease was assessed. The differences in these parameters between the affected area and the nationwide population were analyzed using a chi-square and t-test analysis. A p-value of < 0.05 was considered statistically significant. RESULTS Television was a significantly more common source of information in Israel as a whole (p < 0.05), whereas friends (p < 0.05) and local authorities (p < 0.05) were significantly more common sources of information in the affected area. CONCLUSIONS The frequency of use of the sources of information by the population during the early phase of a bird flu outbreak is different in the affected area compared with the general population in the same country. Authorities must pay attention to this phenomenon and use the correct sources of information in each area in order to achieve better exposure of the population to the recommended behaviors during an outbreak.


title: Novel small-molecule inhibitors of transmissible gastroenteritis virus.

text: We used swine testicle (ST) cells infected with transmissible gastroenteritis virus (TGEV) and an indirect immunofluorescent assay with antibodies against TGEV spike and nucleocapsid proteins to screen small-molecule compounds that inhibit TGEV replication. Analogues of initial hits were collected and subjected to a 3CL protease (3CL(pro)) inhibition assay with recombinant 3CL(pro) and a fluorogenic peptide substrate. A series of benzothiazolium compounds were found to have inhibitory activity against TGEV 3CL(pro) and to exert anti-TGEV activities in terms of viral protein and RNA replication in TGEV-infected ST cells, with consequent protection of TGEV-infected ST cells from cytopathic effect by blocking the activation of caspase-3.


title: Mechanical Thrombectomy in Patients With Acute Ischemic Stroke: A Health Technology Assessment.

text: BACKGROUND In Ontario, current treatment for eligible patients who have an acute ischemic stroke is intravenous thrombolysis (IVT). However, there are some limitations and contraindications to IVT, and outcomes may not be favourable for patients with stroke caused by a proximal intracranial occlusion. An alternative is mechanical thrombectomy with newer devices, and a number of recent studies have suggested that this treatment is more effective for improving functional independence and clinical outcomes. The objective of this health technology assessment was to evaluate the clinical effectiveness and cost-effectiveness of new-generation mechanical thrombectomy devices (with or without IVT) compared to IVT alone (if eligible) in patients with acute ischemic stroke. METHODS We conducted a systematic review of the literature, limited to randomized controlled trials that examined the effectiveness of mechanical thrombectomy using stent retrievers and thromboaspiration devices for patients with acute ischemic stroke. We assessed the quality of the evidence using the GRADE approach. We developed a Markov decision-analytic model to assess the cost-effectiveness of mechanical thrombectomy (with or without IVT) versus IVT alone (if eligible), calculated incremental cost-effectiveness ratios using a 5-year time horizon, and conducted sensitivity analyses to examine the robustness of the estimates. RESULTS There was a substantial, statistically significant difference in rate of functional independence (GRADE: high quality) between those who received mechanical thrombectomy (with or without IVT) and IVT alone (odds ratio [OR] 2.39, 95% confidence interval [CI] 1.88-3.04). We did not observe a difference in mortality (GRADE: moderate quality) (OR 0.80, 95% CI 0.60-1.07) or symptomatic intracerebral hemorrhage (GRADE: moderate quality) (OR 1.11, 95% CI 0.66-1.87). In the base-case cost-utility analysis, which had a 5 year time horizon, the costs and effectiveness for mechanical thrombectomy were $126,939 and 1.484 quality-adjusted life-years (QALYs) (2.969 life-years). The costs and effectiveness for IVT alone were $124,419 and 1.273 QALYs (2.861 life-years), respectively. Mechanical thrombectomy was associated with an incremental cost-effectiveness ratio of $11,990 per QALY gained. Probabilistic sensitivity analysis showed that the probability of mechanical thrombectomy being cost-effective was 57.5%, 89.7%, and 99.6%, at thresholds of $20,000, $50,000, and $100,000 per QALY gained, respectively. We estimated that adopting mechanical thrombectomy would lead to a cost increase of approximately $1 to 2 million. CONCLUSIONS High quality evidence showed that mechanical thrombectomy significantly improved functional independence and appeared to be cost-effective compared to IVT alone for patients with acute ischemic stroke.


Microthrombotic Complications of COVID-19 Are Likely Due to Embolism of Circulating Endothelial Derived Ultralarge Von Willebrand Factor (eULVWF) Decorated-Platelet Strings.


title: Polymorphism of the angiotensin-converting enzyme gene affects the outcome of acute respiratory distress syndrome.

text: OBJECTIVE There has been increasing evidence that angiotensin II may play an important role in the pathogenesis and in the evolution of acute lung injury. It was therefore hypothesized that polymorphisms of the angiotensin-converting enzyme gene affects the risk and outcome of acute respiratory distress syndrome (ARDS). DESIGN Prospective, observational study. PATIENTS AND SETTINGS The ARDS group consisted of 101 patients treated at the medical intensive care unit; the control groups consisted of 138 "at-risk" patients treated at the medical intensive care unit due to acute respiratory failure but did not meet the ARDS criteria throughout the hospital course, and 210 non-at-risk subjects. INTERVENTION None. MEASUREMENTS AND MAIN RESULTS The ARDS patients and control subjects were genotyped for the insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme gene. Association of the polymorphism and the risk and the outcome of ARDS was analyzed. There was no significant difference in the frequencies of the genotypes between the ARDS, at-risk, and non-at-risk groups. The 28-day mortality rates were significantly different between the three angiotensin-converting enzyme genotypes (42%, 65%, and 75% for II, ID, and DD, respectively; p = .036). Survival analysis showed that the II genotype favorably affected 28-day survival (hazard ratio, 0.46; 95% confidence interval, 0.26-0.81; p = .007), whereas ARDS caused by hospital-acquired pneumonia had a negative effect (hazard ratio, 2.34; 95% confidence interval, 1.25-4.40; p = .008). The II genotype (hazard ratio, 0.53; 95% confidence interval, 0.32-0.87; p = .012) and ARDS caused by hospital-acquired pneumonia (hazard ratio, 2.13; 95% confidence interval, 1.24-3.68; p = .006) were also significant prognostic factors for the in-hospital mortality. CONCLUSIONS The angiotensin-converting enzyme I/D polymorphism is a significant prognostic factor for the outcome of ARDS. Patients with the II genotype have a significantly better chance of survival. This study did not show an increased risk for ARDS in Chinese patients with the D allele.


title: Bioactive pyranoxanthones from the roots of Calophyllum blancoi.

text: Phytochemical investigation of the roots of Calophyllum blancoi growing in Taiwan resulted in the isolation of three new pyranoxanthones, blancoxanthone (1), acetyl blancoxanthone (2) and 3-hydroxyblancoxanthone (3), in addition to two known pyranoxanthones, pyranojacaeubin (4) and caloxanthone (5). Structural characterization of the isolated compounds was determined by spectral analyses especially 2-D NMR. Biological study of the isolated xanthones revealed that blancoxanthone (1) exhibited significant anti-coronavirus activity.


title: Avian influenza and animal health risk: conservation of endemic threatened wild birds in Sardinia Island.

text: Sardinia is a Mediterranean island with a long geological history, leading to a separation process from continental Europe during the Miocene. As a consequence, in this insular habitat some wild bird species developed endemic forms, some of which are currently threatened. The aim of this study is to evaluate the possible animal health risk associated with a potential avian influenza virus (AIV) circulation in Sardinian wild bird populations. Overall, 147 cloacal swabs were sampled in the Sardinia region from June 2009 to September 2011. Samples were obtained from 12 taxonomic orders, including 16 families and 40 species of birds. Based on the endangered host status or on the ecology of the host-virus interaction, samples were categorized into three groups of species: 1) endemic, endangered, or both (17 species); 2) potential reservoir (21 species); and 3) potential spillover (two species). Cloacal swabs were tested by reverse transcription (RT)-PCR for influenza A virus matrix gene amplification. Forty-one serum samples were tested by nucleoprotein-enzyme-linked immunosorbent assay (NP-ELISA) for antibodies against influenza A virus nucleoprotein and by hemagglutination inhibition assay for detection of seropositivity against H5 and H7 AIV subtypes. No cloacal swabs tested RT-PCR positive for AIV, whereas two weak seropositive results were detected by NP-ELISA in a mallard (Anas platyrhynchos) and in a yellow-legged gull (Larus michahellis). The low or absent AIV circulation detected in Sardinia's wild birds during the study suggests a naïve status in these avian populations. These data provide new information on AIV circulation in Sardinia's wild birds that could be applied to implement conservation strategies for threatened species.


title: Long-distance RNA-RNA interactions in plant virus gene expression and replication.

text: The vast majority of plant and animal viruses have RNA genomes. Viral gene expression and replication are controlled by cis-acting elements in the viral genome, which have been viewed conventionally as localized structures. However, recent research has altered this perception and provided compelling evidence for cooperative activity involving distantly positioned RNA elements. This chapter focuses on viral RNA elements that interact across hundreds or thousands of intervening nucleotides to control translation, genomic RNA synthesis, and subgenomic mRNA transcription. We discuss evidence supporting the existence and function of the interactions, and speculate on the regulatory roles that such long-distance interactions play in the virus life cycle. We emphasize viruses in the Tombusviridae and Luteoviridae families in which long-distance interactions are best characterized, but similar phenomena in other viruses are also discussed. Many more examples likely remain undiscovered.


Medical technology in the intensive care unit.


title: Evidence of the recombinant origin of a bat severe acute respiratory syndrome (SARS)-like coronavirus and its implications on the direct ancestor of SARS coronavirus.

text: Bats have been identified as the natural reservoir of severe acute respiratory syndrome (SARS)-like and SARS coronaviruses (SLCoV and SCoV). However, previous studies suggested that none of the currently sampled bat SLCoVs is the descendant of the direct ancestor of SCoV, based on their relatively distant phylogenetic relationship. In this study, evidence of the recombinant origin of the genome of a bat SLCoV is demonstrated. We identified a potential recombination breakpoint immediately after the consensus intergenic sequence between open reading frame 1 and the S coding region, suggesting the replication intermediates may participate in the recombination event, as previously speculated for other CoVs. Phylogenetic analysis of its parental regions suggests the presence of an uncharacterized SLCoV lineage that is phylogenetically closer to SCoVs than any of the currently sampled bat SLCoVs. Using various Bayesian molecular-clock models, interspecies transfer of this SLCoV lineage from bats to the amplifying host (e.g., civets) was estimated to have happened a median of 4.08 years before the SARS outbreak. Based on this relatively short window period, we speculate that this uncharacterized SLCoV lineage may contain the direct ancestor of SCoV. This study sheds light on the possible host bat species of the direct ancestor of SCoV, providing valuable information on the scope and focus of surveillance for the origin of SCoV.


title: [Clinical therapy of severe acute respiratory syndrome: 38 cases retrospective analysis].

text: OBJECTIVE To analyze the clinical features and therapy experience of severe acute respiratory syndrome (SARS). METHODS From December 2002 to April 2003 in Guangzhou Institute of Respiratory Disease, 38 patients with severe SARS were retrospectively studied to evaluate the relationship between treatment strategy and prognosis. RESULTS Thirty-eight cases of severe SARS were diagnosed. Comprehensive measures most commonly included corticosteroids, antibiotics, antivirotics, nutritional support and mechanical ventilation. Thirty cases were cured (78.9%), of them 11 cases had pulmonary fibrosis (36.7%), 8 patients died (21.1%) in all cases. CONCLUSION Severe SARS might develop rapidly. In addition to early diagnosis, prompt isolation, and emergency therapy, appropriate use of corticosteroid and noninvasive ventilation should be recommended.


title: Bridging health and foreign policy: the role of health impact assessments.

text: Health impact assessment (HIA) is an important tool for exploring the intersection between health and foreign policy, offering a useful analytical approach to increase positive health impacts and minimize negative impacts. Numerous subject areas have brought health and foreign policy together. Yet further opportunities exist for HIA to address a broader range of health impacts that otherwise may not be seen as relevant to foreign policy. HIA may also improve the quality of scientific evidence available to policy-makers. The Framework Convention on Tobacco Control offers lessons for the strategic use of HIA. However, HIA alone is limited in influencing these decision-making processes, notably when issues diverge from other core concerns such as economics and security. In such cases, HIA is an important tool to be used alongside the mobilization of key constituencies and public support.


title: Hospital preparedness for severe acute respiratory syndrome in the United States: views from a national survey of infectious diseases consultants.

text: In this survey of infectious diseases consultants, 90% reported that their health care facilities have plans in place to address severe acute respiratory syndrome (SARS). Some plan elements exceed current recommendations, whereas others are less stringent. Resource issues associated with airborne isolation and respirators were reported. Sixty-one percent of the respondents expressed some concern about their facility's preparation and capacity for managing patients with SARS. Recent draft guidance on SARS preparedness from the Centers for Disease Control and Prevention may help address some of these issues.


title: Activation of the envelope proteins by a metalloproteinase enables attachment and entry of the hepatitis B virus into T-lymphocyte.

text: Previously, we identified an HBV binding factor (HBV-BF), a 50-kDa serum glycoprotein which interacts with HBV envelope proteins and which is also located in the membrane of normal human hepatocyte (A. Budkowska et al. (1993) J. Virol. 67, 4316). Here we show that HBV-BF is a neutral metalloproteinase which shares substrate specificity and properties with a newly described family of membrane type matrix metalloproteinases. HBV-BF treatment of the HBV resulted in the cleavage of the N-terminal part of the middle HBV envelope protein at the pre-S2(136-141) amino acid sequence VRGLYF/L (containing a single arginine cleavage site). HBV-BF affected the reactivity of the large HBV protein with pre-S1-specific MAbs, probably inducing the conformational change of the pre-S1 domain. The HBV-BF-digested virus remained morphologically intact with unchanged S antigenic determinants. The structural modifications of the viral envelope proteins induced by HBV-BF enabled cell membrane attachment and viral entry into the T-lymphocyte. Both processes were blocked by the metalloproteinase inhibitor 1,10 phenanthroline. Thus, the host-dependent proteolytic activation of the envelope proteins seems to be essential for the HBV entry into the cell. HBV-BF under a membrane bound or a secreted form could be (one of) the molecule(s) responsible for the HBV proteolytic activation.


title: Characterizing Transmission and Control of the SARS Epidemic: Novel Stochastic Spatio-Temporal Models.

text: Severe Acute Respiratory Syndrome (SARS), the first epidemic of the 21st century, has an outbreak history of more than 2 years till today and caused tremendous damage to the human society. Accordingly, many studies on modeling the SARS epidemic have been reported, whereas deficiencies were still lying in those models because of their separate space/time methodology. In this paper, we propose novel comprehensive stochastic spatio-temporal models from both of the macro aspect and individual aspect for characterizing transmission and control of the SARS disease. Based on a new SARS spread process in consideration of "suspicious" population, we firstly establish the stochastic temporal models from two different aspects: the macro model is described with birth-death process and the individual Markov model is described with probability transition matrix (PTM). And then, we amalgamate the deterministic/stochastic population-flow model with the stochastic temporal models together to set up the comprehensive stochastic spatio-temporal models. Simulations on computer have evaluated the effect of various realistic parameters and control policies, and also have testified the accuracy and efficacy of the new models. Additionally, particular studies on the cases of Tsinghua University and Beijing City are presented. The comprehensive stochastic spatio-temporal models have considerably reduced the complexity plus errors as compared with previous works and will be able to characterize other various epidemics, e.g. Avian Flu.


title: Is humidification always necessary during noninvasive ventilation in the hospital?

text: Noninvasive ventilation (NIV) is a standard of care for the treatment of exacerbation of chronic obstructive pulmonary disease, to prevent intubation and reduce morbidity and mortality. The need for humidification of NIV gas is controversial. Some unique aspects of NIV conspire to alter the delivered humidity and airway function. In the presence of air leaks, unidirectional air flow dries the airways and increases airway resistance. Patient comfort is also a critical issue, as tolerance of NIV is often tied to patient comfort. This paper provides the arguments for and against routine humidification during NIV in the hospital setting. Data from clinical research demonstrate the effects of delivered humidification on relevant physiologic variables. The impact of humidification on NIV success/failure remains speculative.


title: Human immunodeficiency virus-1-infected macrophages induce inducible nitric oxide synthase and nitric oxide (NO) production in astrocytes: astrocytic NO as a possible mediator of neural damage in acquired immunodeficiency syndrome.

text: Nitric oxide (NO) plays an important role in normal neural cell function. Dysregulated or overexpression of NO contributes to neurologic damage associated with various pathologies, including human immunodeficiency virus (HIV)-associated neurological disease. Previous studies suggest that HIV-infected monocyte-derived macrophages (MDM) produce low levels of NO in vitro and that inducible nitric oxide synthase (iNOS) is expressed in the brain of patients with neurologic disease. However, the levels of NO could not account for the degree of neural toxicity observed. In this study, we found that induction of iNOS with concomitant production of NO occurred in primary human astrocytes, but not in MDM, when astrocytes were cocultured with HIV-1-infected MDM. This coincided with decreased HIV replication in infected MDM. Supernatants from cocultures of infected MDM and astrocytes also stimulated iNOS/NO expression in astrocytes, but cytokines known to induce iNOS expression (interferon-gamma, interleukin-1beta, and tumor necrosis factor-alpha) were not detected. In addition, the recombinant HIV-1 envelope protein gp41, but not rgp120, induced iNOS in cocultures of uninfected MDM and astrocytes. This suggests that astrocytes may be an important source of NO production due to dysregulated iNOS expression and may constitute one arm of the host response resulting in suppression of HIV-1 replication in the brain. It also leads us to speculate that neurologic damage observed in HIV disease may ensue from prolonged, high level production of NO.


title: RFID in healthcare: a Six Sigma DMAIC and simulation case study.

text: PURPOSE The purpose of this paper is to develop a business model to generate quantitative evidence of the benefits of implementing radio frequency identification (RFID) technology, limiting the scope to outpatient surgical processes in hospitals. DESIGN/METHODOLOGY/APPROACH The study primarily uses the define-measure-analyze-improve-control (DMAIC) approach, and draws on various analytical tools such as work flow diagrams, value stream mapping, and discrete event simulation to examine the effect of implementing RFID technology on improving effectiveness (quality and timeliness) and efficiency (cost reduction) of outpatient surgical processes. FINDINGS The analysis showed significant estimated annual cost and time savings in carrying out patients' surgical procedures with RFID technology implementation for the outpatient surgery processes in a hospital. This is largely due to the elimination of both non-value added activities of locating supplies and equipment and also the elimination of the "return" loop created by preventable post operative infections. Several poka-yokes developed using RFID technology were identified to eliminate those two issues. PRACTICAL IMPLICATIONS Several poka-yokes developed using RFID technology were identified for improving the safety of the patient and cost effectiveness of the operation to ensure the success of the outpatient surgical process. ORIGINALITY/VALUE Many stakeholders in the hospital environment will be impacted including patients, physicians, nurses, technicians, administrators and other hospital personnel. Different levels of training of hospital personnel will be required, based on the degree of interaction with the RFID system. Computations of costs and savings will help decision makers understand the benefits and implications of the technology in the hospital environment.


title: Economically important non-oncogenic immunosuppressive viral diseases of chicken--current status.

text: Immunosuppressive viral diseases threaten the poultry industry by causing heavy mortality and economic loss of production, often as a result of the chickens' increased susceptibility to secondary infections and sub-optimal response to vaccinations. This paper aimed to present an up-to-date review of three specific economically important non-oncogenic immunosuppressive viral diseases of chickens, viz. chicken infectious anaemia (CIA), infectious bursal disease (IBD) and hydropericardium syndrome (HPS), with emphasis on their immunosuppressive effects. CIA and IBD causes immunosuppression in chickens and the socio-economic significance of these diseases is considerable worldwide. CIA occurs following transovarian transmission of chicken anaemia virus and has potential for inducing immunosuppression alone or in combination with other infectious agents, and is characterized by generalized lymphoid atrophy, increased mortality and severe anemia. The virus replicates in erythroid and lymphoid progenitor cells, causing inapparent, sub-clinical infections that lead to depletion of these cells with consequent immunosuppressive effects. The IBD virus replicates extensively in IgM(+) cells of the bursa and chickens may die during the acute phase of the disease, although IBD virus-induced mortality is highly variable and depends, among other factors, upon the virulence of the virus strain. The sub-clinical form is more common than clinical IBD because of regular vaccination on breeding farms. Infection at an early age significantly compromises the humoral and local immune responses of chickens because of the direct effect of B cells or their precursors. HPS is a recently emerged immunosuppressive disease of 3-6-weeked broilers, characterized by sudden onset, high mortality, typical hydropericardium and enlarged mottled and friable livers, with intranuclear inclusion bodies in the hepatocytes. The agent, fowl adenovirus-4, causes immunosuppression by damaging lymphoid tissues; the presence of IBD and CIA viruses may predispose for HPS or HPS may predispose for other viral infections. Synergism with CIA or other virus infections or prior immunosuppression is necessary to produce IBH-HPS in chickens and the susceptibility of chickens infected with fowl adenovirus varies throughout the course of CIA infection. The mechanism of immunosuppression has been studied in detail for certain chicken viruses at molecular levels, which will provides new opportunities to control these diseases by vaccination.


Ribavirin in the treatment of severe acute respiratory syndrome (SARS).


title: COVID-19-CT-CXR: a freely accessible and weakly labeled chest X-ray and CT image collection on COVID-19 from biomedical literature.

text: The latest threat to global health is the COVID-19 outbreak. Although there exist large datasets of chest X-rays (CXR) and computed tomography (CT) scans, few COVID-19 image collections are currently available due to patient privacy. At the same time, there is a rapid growth of COVID-19-relevant articles in the biomedical literature. Here, we present COVID-19-CT-CXR, a public database of COVID-19 CXR and CT images, which are automatically extracted from COVID-19-relevant articles from the PubMed Central Open Access (PMC-OA) Subset. We extracted figures, associated captions, and relevant figure descriptions in the article and separated compound figures into subfigures. We also designed a deep-learning model to distinguish them from other figure types and to classify them accordingly. The final database includes 1,327 CT and 263 CXR images (as of May 9, 2020) with their relevant text. To demonstrate the utility of COVID-19-CT-CXR, we conducted four case studies. (1) We show that COVID-19-CT-CXR, when used as additional training data, is able to contribute to improved DL performance for the classification of COVID-19 and non-COVID-19 CT. (2) We collected CT images of influenza and trained a DL baseline to distinguish a diagnosis of COVID-19, influenza, or normal or other types of diseases on CT. (3) We trained an unsupervised one-class classifier from non-COVID-19 CXR and performed anomaly detection to detect COVID-19 CXR. (4) From text-mined captions and figure descriptions, we compared clinical symptoms and clinical findings of COVID-19 vs. those of influenza to demonstrate the disease differences in the scientific publications. We believe that our work is complementary to existing resources and hope that it will contribute to medical image analysis of the COVID-19 pandemic. The dataset, code, and DL models are publicly available at https://github.com/ncbi-nlp/COVID-19-CT-CXR.


title: Exciting times: towards a totally minimally invasive paediatric urology service.

text: Following on from the first paediatric laparoscopic nephrectomy in 1992, the growth of minimally invasive ablative and reconstructive procedures in paediatric urology has been dramatic. This article reviews the literature related to laparoscopic dismembered pyeloplasty, optimising posterior urethral valve ablation and intravesical laparoscopic ureteric reimplantation.


title: Tracking the evolution of the SARS coronavirus using high-throughput, high-density resequencing arrays.

text: Mutations in the SARS-Coronavirus (SARS-CoV) can alter its clinical presentation, and the study of its mutation patterns in human populations can facilitate contact tracing. Here, we describe the development and validation of an oligonucleotide resequencing array for interrogating the entire 30-kb SARS-CoV genome in a rapid, cost-effective fashion. Using this platform, we sequenced SARS-CoV genomes from Vero cell culture isolates of 12 patients and directly from four patient tissues. The sequence obtained from the array is highly reproducible, accurate (>99.99% accuracy) and capable of identifying known and novel variants of SARS-CoV. Notably, we applied this technology to a field specimen of probable SARS and rapidly deduced its infectious source. We demonstrate that array-based resequencing-by-hybridization is a fast, reliable, and economical alternative to capillary sequencing for obtaining SARS-CoV genomic sequence on a population scale, making this an ideal platform for the global monitoring of SARS-CoV and other small-genome pathogens.


title: [Hepatocyte transplantation].

text: BACKGROUND Transplantation of hepatocytes is considered a promising technology for the cell therapy of liver diseases. Cells are isolated from donor livers, which are not allocated for organ transplantation and transplanted into the liver of a suitable recipient. Ideally, the transplanted cells functionally replace the hepatocytes of the diseased organ and restore its metabolic capacity either permanently or for a period of bridging to organ transplantation. METHODS AND RESULTS Although about 50 cases of clinical hepatocyte transplantation have been documented, therapeutic benefit is doubtful, and the reasons for this are largely unknown. Minor quality of the transplanted cells isolated from marginal donor livers may be a major cause. Therefore, animal models have been established, which enable scientists to develop novel procedures of hepatocyte transplantation and to specifically study the mechanisms of hepatocyte integration in the host liver. Hopefully, results generated in animal models will set the basis for the design of novel procedures of hepatocyte transplantation individualized to the underlying disease in order to provide a growth advantage for donor over host cells. PERSPECTIVES The plasticity of stem cells and their proliferative potential is the basis for current efforts to generate stem cell-derived hepatocytes of transplantation quality for the treatment of liver diseases.


title: Nasal continuous positive airway pressure (CPAP) for the respiratory care of the newborn infant.

text: Nasal continuous positive airway pressure (CPAP) is a noninvasive form of respiratory assistance that has been used to support spontaneously breathing infants with lung disease for nearly 40 years. Following reports that mechanical ventilation contributes to pulmonary growth arrest and the development of chronic lung disease, there is a renewed interest in using CPAP as the prevailing method for supporting newborn infants. Animal and human research has shown that CPAP is less injurious to the lungs than is mechanical ventilation. The major concepts that embrace lung protection during CPAP are the application of spontaneous breathing at a constant distending pressure and avoidance of intubation and positive-pressure inflations. A major topic for current research focuses on whether premature infants should be supported initially with CPAP following delivery, or after the infant has been extubated following prophylactic surfactant administration. Clinical trials have shown that CPAP reduces the need for intubation/mechanical ventilation and surfactant administration, but it is still unclear whether CPAP reduces chronic lung disease and mortality, compared to modern lung-protective ventilation techniques. Despite the successes, little is known about how best to manage patients using CPAP. It is also unclear whether different strategies or devices used to maintain CPAP play a role in improving outcomes in infants. Nasal CPAP technology has evolved over the last 10 years, and bench and clinical research has evaluated differences in physiologic effects related to these new devices. Ultimately, clinicians' abilities to perceive changes in the pathophysiologic conditions of infants receiving CPAP and the quality of airway care provided are likely to be the most influential factors in determining patient outcomes.


title: Video-assisted thoracic surgery for left upper lobectomy for complex lesions: how to extend the indication with optimal safety?

text: The feasibility of extending the VATS approach to locally advanced NSCLC has been described with good clinical outcome. These complex resections are still technically challenging and patient safety must remain the highest priority. In this article, we describe our routine VATS approach for left upper lobectomy in proximal, locally advanced lesions. Both surgical and anaesthesiology teams are trained during simulation sessions to respond rapidly in case of urgent thoracotomy. Encircling arterial and venous vessels allow control of inadvertent bleeding during difficult dissection. Also, whenever needed the double vessel control technique is a time saver waiting for conversion to thoracotomy.


title: Tissue-specific pattern of angiotensin-converting enzyme 2 expression in rat pancreas.

text: Angiotensin-converting enzyme 2 (ACE2), a carboxypeptidase that is highly homologous to ACE, acts as a negative regulator for the renin-angiotensin system (RAS). Pancreatic RAS is thought to play important endocrine and exocrine roles in hormone secretion. Further exploration of this system is likely to offer new insights into the pathogenesis of pancreatic diseases such as diabetes mellitus. This study investigated the expression of ACE2 in rat pancreatic exocrine and endocrine tissue. Reverse transcription-polymerase chain reaction, immunoblotting and immunohistochemistry showed that ACE2 mRNA and protein were expressed in the pancreas. Immunoelectron microscopy demonstrated that ACE2 was expressed in both endocrine and exocrine pancreatic tissues. In the endocrine tissue, ACE2 was localized on the secretory granules. Double immunofluorescence labelling showed that ACE2 was co-localized with both insulin and somatostatin, while it was rarely co-localized with glucagon and pancreatic polypeptide. These findings suggest that ACE2 might play an important role in glucose homeostasis.


title: Design, synthesis and antiviral activity of 2-(3-amino-4-piperazinylphenyl)chromone derivatives.

text: Previously, we have confirmed that the antiviral activities of the chromone derivatives were controlled by the type as well as the position of the substituents attached to the chromone core structure. In the course of our ongoing efforts to optimize the antiviral activity of the chromone derivatives, we have been attempting to derivatize the chromone scaffold via introduction of various substituents. In this proof-of-concept study, we introduced a 3-amino-4-piperazinylphenyl functionality to the chromone scaffold and evaluated the antiviral activities of the resulting chromone derivatives. The synthesized 2-(3-amino-4-piperazinylphenyl)-chromones showed severe acute respiratory syndrome-corona virus (SARS-CoV)-specific antiviral activity. In particular, the 2-pyridinylpiperazinylphenyl substituents provided the resulting chromone derivatives with selective antiviral activity. Taken together, this result indicates the possible pharmacophoric role of the 2-pyridinylpiperazine functionality attached to the chromone scaffold, which warrants further in-depth structure-activity relationship study.


title: Novel therapeutic vaccines [(HSP65 + IL-12)DNA-, granulysin- and Ksp37-vaccine] against tuberculosis and synergistic effects in the combination with chemotherapy.

text: PURPOSE Multi-drug resistant tuberculosis (MDR-TB) and extremely drug resistant (XDR) TB are big problems in the world. We have developed novel TB therapeutic vaccines, HVJ-Envelope/HSP65 + IL-12 DNA vaccine (HSP65-vaccine), granulysin vaccine and killer specific secretory protein of 37kDa (Ksp37) vaccine. METHODS AND RESULTS HSP65 vaccine showed strong therapeutic effect against both MDR-TB and XDR-TB in mice. Intradermal immunization of HSP65-vaccine showed stronger therapeutic effect against TB than intramuscular or subcutaneous immunization. Furthermore, the synergistic therapeutic effect was observed when the vaccine was administrated in combination with Isoniazid (INH), which is a first line drug for chemotherapy. The combination of types of vaccines (HSP65- and granulysin- vaccines) also showed synergistic therapeutic effect. In the monkey model, granulysin-vaccine prolonged the survival period after the infection of TB and long-term survival was observed in vaccine-treated group. We examined the potential of two kinds of novel DNA vaccines (Ksp37-vaccine and granulysin-vaccine). Both vaccines augmented in vivo differentiation of CTL against TB. We measured the amount of Ksp37 protein in human serum and revealed that the level of Ksp37 protein of patients with tuberculosis was lower than that of healthy volunteers. Therefore, we established Ksp37 transgenic mice as well as granulysin transgenic mice to elucidate the function of those proteins. Both transgenic mice were resistant to TB infection. CONCLUSION These data indicate the potential of combinational therapy; the combination of two DNA vaccines or combination of DNA vaccine with antibiotic drug. Thus, it will provide a novel strategy for the treatment of MDR-TB.


title: Predominance of Th2 cytokines, CXC chemokines and innate immunity mediators at the mucosal level during severe respiratory syncytial virus infection in children.

text: UNLABELLED Profiling of immune mediators in both nasal and plasma samples is a common approach to the study of pathogenesis in respiratory viral infections. Nevertheless, mucosal immunity functions essentially independently from peripheral immunity. In our study, 27 immune mediators were profiled in parallel, in nasopharyngeal aspirates (NPAs) and plasma from 22 < 2 year-old children with a severe respiratory syncytial virus infection involving the lower respiratory tract, using a multiplex assay. NPAs from 22 children with innocent heart murmurs were used as controls. Differences in mediator concentrations between NPAs from patients and controls were assessed using the Mann-Whitney test. Ratios of innate/adaptive-immunity mediators, Th2/Th1-cytokines and CXC/CC-chemokines were calculated for NPAs and plasmas and differences were assessed using the Wilcoxon test. Associations mediators, severity and leukocyte counts were studied using the Spearman-Karber test. RESULTS increased levels of Th1 cytokines (IL-1beta, IL-2, IL-12p70, IFNgamma, TNFalpha), Th2 cytokines (IL-13, IL-4, IL-6, IL-10), chemokines (IP-10, IL-8, MIP1alpha, MIP-1beta), growth factors (FGFb, PDGFbb, GCSF) and IL-1RA, IL-17 were observed in patient NPAs in comparison to controls. In the relative comparisons between patient NPAs and plasmas, a predominance of innate immunity mediators, Th2 cytokines and CXC chemokines was found at the mucosal level. No association between the level of each mediator in NPAs and plasma was found. In plasma, PDGFbb, VEGF, MIP-1alpha, IL-8 correlated with severity; RANTES and IL-6 correlated with leukocyte counts. CONCLUSIONS acute respiratory syncytial virus infection induces a relative predominance of innate-immunity mediators, Th2 cytokines and CXC chemokines in the mucosal compartment in infected children.


Medical Students Contribute to the Community and Medical Field During COVID-19 Pandemic.


title: Programmed -1 ribosomal frameshifting in the SARS coronavirus.

text: Programmed -1 ribosomal frameshifting is an alternate mechanism of translation used by coronavirus to synthesize replication proteins encoded by two overlapping open reading frames. For some coronaviruses, the mRNA cis-acting stimulatory structures involved in this process have been characterized, but their precise contribution to ribosomal frameshifting is not completely understood. Recently, a novel coronavirus was identified as the causative agent of the severe acute respiratory syndrome. This review describes the mRNA motifs involved in programmed -1 ribosomal frameshifting in this virus.


title: Identification and characterization of the putative fusion peptide of the severe acute respiratory syndrome-associated coronavirus spike protein.

text: Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is a newly identified member of the family Coronaviridae and poses a serious public health threat. Recent studies indicated that the SARS-CoV viral spike glycoprotein is a class I viral fusion protein. A fusion peptide present at the N-terminal region of class I viral fusion proteins is believed to initiate viral and cell membrane interactions and subsequent fusion. Although the SARS-CoV fusion protein heptad repeats have been well characterized, the fusion peptide has yet to be identified. Based on the conserved features of known viral fusion peptides and using Wimley and White interfacial hydrophobicity plots, we have identified two putative fusion peptides (SARS(WW-I) and SARS(WW-II)) at the N terminus of the SARS-CoV S2 subunit. Both peptides are hydrophobic and rich in alanine, glycine, and/or phenylalanine residues and contain a canonical fusion tripeptide along with a central proline residue. Only the SARS(WW-I) peptide strongly partitioned into the membranes of large unilamellar vesicles (LUV), adopting a beta-sheet structure. Likewise, only SARS(WW-I) induced the fusion of LUV and caused membrane leakage of vesicle contents at peptide/lipid ratios of 1:50 and 1:100, respectively. The activity of this synthetic peptide appeared to be dependent on its amino acid (aa) sequence, as scrambling the peptide rendered it unable to partition into LUV, assume a defined secondary structure, or induce both fusion and leakage of LUV. Based on the activity of SARS(WW-I), we propose that the hydrophobic stretch of 19 aa corresponding to residues 770 to 788 is a fusion peptide of the SARS-CoV S2 subunit.


title: Current molecular techniques for the detection of microbial pathogens.

text: Traditionally the detection of microbial pathogens in clinical, environmental or food samples has commonly needed the prelevation of cells by culture before the application ofthe detection strategy. This is done to increase cell number thereby overcoming problems associated with the sensitivity of classical detection strategies. However, culture-based methods have the disadvantages of taking longer, usually are more complex and require skilled personnel as well as not being able to detect viable but non cultivable microbial species. A number of molecular methods have been developed in the last 10 to 15 years to overcome these issues and to facilitate the rapid, accurate, sensitive and cost effective identification and enumeration of microorganisms which are designed to replace and/or support classical approaches to microbial detection. Amongst these new methods, ones based on the polymerase chain reaction and nucleic acid hybridization have been shown to be particularly suitable for this purpose. This review generally summarizes some of the current and emerging nucleic acid based molecular approaches for the detection, discrimination andquantification ofmicrobes in environmental, food and clinical samples and includes reference to the recently developing areas of microfluidics and nanotechnology "Lab-on-a-chip".


title: New partnerships between animal health services and public health agencies.

text: As Veterinary Services and animal health organisations attempt to respond to a new era of emerging and re-emerging zoonotic diseases, their ability and skill in forming new strategic partnerships will be paramount. While these new partnerships are likely to include many relationships outside traditional Veterinary Services and animal agriculture, none will become more important than the formation of new animal health and public health partnerships. Episodes of emerging zoonoses are being increasingly recognised around the world and the confluence of people, animals and animal products today is unprecedented. Concurrently, a wide array of complex factors are also converging that will not only ensure the continuous emergence of zoonoses, but are also likely to drive the further increase and expansion of these diseases. This article discusses the need for the creation of more effective and co-operative partnerships in the face of new microbial threats, the complexity of both the formation and expansion of zoonoses, and the collective abilities of both human and animal health services to respond to them. Lessons learned from recent zoonotic epidemics supportthe need for co-ordinated research, interdisciplinary centres, integrated surveillance systems, response systems and infrastructures, and workforce development strategies. While there are some excellent examples of collaborative animal and public health relationships, there is no question that more and stronger partnerships among national and international organisations, both academic and private, will be necessary to meet the future challenges of emerging zoonoses and to manage their profound implications.


The classification and nomenclature of viruses. Summary of results of meetings of the International Committee on Taxonomy of Viruses in Madrid, September 1975.


title: Neutralizing human antibodies to varicella-zoster virus (VZV) derived from a VZV patient recombinant antibody library.

text: Varicella-zoster virus (VZV), the causative agent of chickenpox and herpes zoster, can be life-threatening in prematurely born children and in children with immune defects or who are under immunosuppressive treatment. Therefore agents for passive immunization, such as VZV-specific immunoglobulin preparations (VZIG) derived from convalescent plasma, are crucial in the prophylaxis of VZV infection. This study describes the isolation of human VZV-neutralizing recombinant antibodies. A human single-chain variable fragment (scFv) phage display library was generated from RNA extracted from peripheral blood lymphocytes of a convalescent varicella patient. Specific phage antibodies were selected against VZV-infected human fibroblasts, and eight unique clones were further expressed as soluble scFv in Escherichia coli. They all showed binding characteristics to varicella antigens with affinities in the K(D) range 0.1-0.2 muM. Two of the scFv antibodies, VZV4 and VZV5, showed dose-dependent in vitro neutralization of VZV. VZV39 also showed a neutralizing effect as scFv, an effect that was increased 4000-fold by conversion into IgG and was further increased by the addition of complement. This is possibly the first time that monovalent scFv antibodies have been shown to neutralize VZV in vitro. This finding will have an impact on the production of new prophylactic antibodies, as such antibody fragments can be cost-effectively produced in E. coli. The antibodies isolated bind both complement-dependent and -independent epitopes for neutralization, thus they may prove useful tools for the study of VZV virulence mechanisms.


title: Differential expression of neuronal ACE2 in transgenic mice with overexpression of the brain renin-angiotensin system.

text: Angiotensin-converting enzyme 2 (ACE2) is a newly discovered carboxy-peptidase responsible for the formation of vasodilatory peptides such as angiotensin-(1-7). We hypothesized that ACE2 is part of the brain renin-angiotensin system, and its expression is regulated by the other elements of this system. ACE2 immunostaining was performed in transgenic mouse brain sections from neuron-specific enolase-AT(1A) (overexpressing AT(1A) receptors), R(+)A(+) (overexpressing angiotensinogen and renin), and control (nontransgenic littermates) mice. Results show that ACE2 staining is widely distributed throughout the brain. Using cell-type-specific antibodies, we observed that ACE2 staining is present in the cytoplasm of neuronal cell bodies but not in glial cells. In the subfornical organ, an area lacking the blood-brain barrier and sensitive to blood-borne angiotensin II, ACE2 was significantly increased in transgenic mice. Interestingly, ACE2 mRNA and protein expression were inversely correlated in the nucleus of tractus solitarius/dorsal motor nucleus of the vagus and the ventrolateral medulla, when comparing transgenic to nontransgenic mice. These results suggest that ACE2 is localized to the cytoplasm of neuronal cells in the brain and that ACE2 levels appear highly regulated by other components of the renin-angiotensin system, confirming its involvement in this system. Moreover, ACE2 expression in brain structures involved in the control of cardiovascular function suggests that the carboxypeptidase may have a role in the central regulation of blood pressure and diseases involving the autonomic nervous system, such as hypertension.


title: Effects of subchronic inhalation exposure of rats to emissions from a diesel engine burning soybean oil-derived biodiesel fuel.

text: There is increasing interest in diesel fuels derived from plant oils or animal fats ("biodiesel"), but little information on the toxicity of biodiesel emissions other than bacterial mutagenicity. F344 rats were exposed by inhalation 6 h/day, 5 days/wk for 13 wk to 1 of 3 dilutions of emissions from a diesel engine burning 100% soybean oil-derived fuel, or to clean air as controls. Whole emissions were diluted to nominal NO(x) concentrations of 5, 25, or 50 ppm, corresponding to approximately 0.04, 0.2, and 0.5 mg particles/m(3), respectively. Biologically significant, exposure-related effects were limited to the lung, were greater in females than in males, and were observed primarily at the highest exposure level. There was a dose-related increase in the numbers of alveolar macrophages and the numbers of particles in the macrophages, as expected from repeated exposure, but no neutrophil response even at the highest exposure level. The macrophage response was reduced 28 days after cessation of the exposure. Among the high-level females, the group mean lung weight/body weight ratio was increased, and minimal, multifocal bronchiolar metaplasia of alveolar ducts was observed in 4 of 30 rats. Lung weights were not significantly increased, and metaplasia of the alveolar ducts was not observed in males. An increase in particle-laden macrophages was the only exposure-related finding in lungs at the intermediate and low levels, with fewer macrophages and fewer particles per macrophage at the low level. Alveolar histiocytosis was observed in a few rats in both exposed and control groups. There were statistically significant, but minor and not consistently exposure-related, differences in body weight, nonpulmonary organ weights, serum chemistry, and glial fibrillary acidic protein in the brain. There were no significant exposure-related effects on survival, clinical signs, feed consumption, ocular toxicity, hematology, neurohistology, micronuclei in bone marrow, sister chromatid exchanges in peripheral blood lymphocytes, fertility, reproductive toxicity, or teratology. This study demonstrated modest adverse effects at the highest exposure level, and none other than the expected physiological macrophage response to repeated particle exposure at the intermediate level.


Poult enteritis and mortality syndrome in turkeys in Great Britain.


title: Identification of Dengue-specific B-Cell Epitopes by Phage-display Random Peptide Library.

text: BACKGROUND Dengue is the most important human viral disease transmitted by arthropod vectors. The availability of random peptide libraries (RPL) displayed on phage has provided a powerful tool for selecting sequences that mimic epitopes from microorganisms that are useful for diagnostic and vaccine development purposes. In this paper, we describe peptides that resemble the antigenic structure of B-cell epitopes of dengue virus identified from a phage-peptide library using human sera containing polyclonal antibodies against dengue virus. MATERIALS AND METHODS Eighteen phage clones were isolated from the phage-display peptide library, J404, by affinity selection using human antisera against dengue virus type 3. These clones were tested for reactivity by ELISA with a panel of hyperimmune ascitic fluids (HAFs) containing antibodies either against all four dengue serotypes, West Nile virus (WNV) or Eastern equine encephalitis virus (EEEV) with control ascitic fluid (NAF) used as a negative control. RESULTS Eight clones were recognized by HAFs against the four dengue serotypes, of which four significantly inhibited binding of anti-dengue antibodies to the virus. Two peptides with similar sequences to regions of NS3 and NS4B non-structural dengue virus proteins were identified. CONCLUSION Our results suggest that these peptides could be used for the development of diagnostic tools for the detection of dengue virus infection and for a potential vaccine against this pathogen.


Tsunami aid spending draws fire over public health.


title: Relevance of nucleic acid amplification techniques for diagnosis of respiratory tract infections in the clinical laboratory.

text: Clinical laboratories are increasingly receiving requests to perform nucleic acid amplification tests for the detection of a wide variety of infectious agents. In this paper, the efficiency of nucleic acid amplification techniques for the diagnosis of respiratory tract infections is reviewed. In general, these techniques should be applied only for the detection of microorganisms for which available diagnostic techniques are markedly insensitive or nonexistent or when turnaround times for existing tests (e.g., viral culture) are much longer than those expected with amplification. This is the case for rhinoviruses, coronaviruses, and hantaviruses causing a pulmonary syndrome, Bordetella pertussis, Chlamydia pneumoniae, Mycoplasma pneumoniae, and Coxiella burnetii. For Legionella spp. and fungi, contamination originating from the environment is a limiting factor in interpretation of results, as is the difficulty in differentiating colonization and infection. Detection of these agents in urine or blood by amplification techniques remains to be evaluated. In the clinical setting, there is no need for molecular diagnostic tests for the diagnosis of Pneumocystis carinii. At present, amplification methods for Mycobacterium tuberculosis cannot replace the classical diagnostic techniques, due to their lack of sensitivity and the absence of specific internal controls for the detection of inhibitors of the reaction. Also, the results of interlaboratory comparisons are unsatisfactory. Furthermore, isolates are needed for susceptibility studies. Additional work remains to be done on sample preparation methods, comparison between different amplification methods, and analysis of results. The techniques can be useful for the rapid identification of M. tuberculosis in particular circumstances, as well as the rapid detection of most rifampin-resistant isolates. The introduction of diagnostic amplification techniques into a clinical laboratory implies a level of proficiency for excluding false-positive and false-negative results.


title: Emerging viral infections in transplant recipients.

text: PURPOSE OF REVIEW Transplant patients are uniquely predisposed to emerging infections for a number of reasons. Two outbreaks, West Nile virus and severe acute respiratory syndrome, have recently provided important lessons on how transplant patients are affected, and how transplant programmes must adapt and evolve in the face of emerging infections. An update of emerging infections in transplant patients, using West Nile virus and severe acute respiratory syndrome as specific examples, is summarized here. RECENT FINDINGS Exogenous immunosuppression, specific allograft factors, and extensive contact with the healthcare system all predispose transplant patients to emerging infections. Transplant patients may acquire West Nile virus through blood transfusion, donor transmission, or community exposure. Seroprevalence data in transplant populations suggest the risk of severe neurological disease is several fold higher in transplant recipients who acquire West Nile virus compared with immunocompetent individuals. Prevention strategies are critical in this population. These include nucleic acid testing of blood products and potentially also screening organ donors in a similar manner. During the outbreak of severe acute respiratory syndrome, transplant patients with severe and rapidly progressive disease were reported. Higher viral burdens appeared to be present in transplant patients and may have implications for the increased infectivity of these patients. Transplant programmes in severe acute respiratory syndrome areas were also adversely affected because of donor concerns, recipient issues and resource problems. SUMMARY Transplant patients are uniquely predisposed to emerging infections. Lessons learned from West Nile virus and severe acute respiratory syndrome in transplantation should be applicable to future outbreaks of other emerging infectious diseases.


title: Psychobehavioural responses to the 2014 Middle East respiratory syndrome-novel corona virus (MERS CoV) among adults in two shopping malls in Jeddah, western Saudi Arabia.

text: Sporadic cases of Middle East respiratory syndrome caused by a novel corona virus (MERS-CoV) were first detected in Saudi Arabia in June 2012. The number of cases was highest during April and May 2014. To assess determinants of psychobehavioural responses among the general population in Jeddah, western Saudi Arabia, a cross-sectional survey was conducted at the end of June 2014. Data included sociodemographic characteristics, level of anxiety, protective measures and social avoidance responses. A total of 358 participants completed the questionnaire; 58.4% were female, and the age range was 18-72 years. None of the participants was diagnosed with MERS-CoV. More than half (57.7%) recorded a moderate anxiety score using a visual analogue scale. Anxiety level was significantly associated with increased perception of susceptibility to infection and social avoidance behaviours related to travel and being in public places.


title: Evidence for coronavirus discontinuous transcription.

text: Coronavirus subgenomic mRNA possesses a 5'-end leader sequence which is derived from the 5' end of genomic RNA and is linked to the mRNA body sequence. This study examined whether coronavirus transcription involves a discontinuous transcription step; the possibility that a leader sequence from mouse hepatitis virus (MHV) genomic RNA could be used for MHV subgenomic defective interfering (DI) RNA transcription was examined. This was tested by using helper viruses and DI RNAs that were easily distinguishable. MHV JHM variant JHM(2), which synthesizes a subgenomic mRNA encoding the HE gene, and variant JHM(3-9), which does not synthesize this mRNA, were used. An MHV DI RNA, DI(J3-9), was constructed to contain a JHM(3-9)-derived leader sequence and an inserted intergenic region derived from the region preceding the MHV JHM HE gene. DI(J3-9) replicated efficiently in JHM(2)- or JHM(3-9)-infected cells, whereas synthesis of subgenomic DI RNAs was observed only in JHM(2)-infected cells. Sequence analyses demonstrated that the 5' regions of both helper virus genomic RNAs and genomic DI RNAs maintained their original sequences in DI RNA-replicating cells, indicating that the genomic leader sequences derived from JHM(2) functioned for subgenomic DI RNA transcription. Replication and transcription of DI(J3-9) were observed in cells infected with an MHV A59 strain whose leader sequence was similar to that of JHM(2), except for one nucleotide substitution within the leader sequence. The 5' region of the helper virus genomic RNA and that of the DI RNA were the same as their original structures in virus-infected cells, and the leader sequence of DI(J3-9) subgenomic DI RNA contained the MHV A59-derived leader sequence. The leader sequence of subgenomic DI RNA was derived from that of helper virus; therefore, the genomic leader sequence had a trans-acting property indicative of a discontinuous step in coronavirus transcription.


Care of the well newborn.


title: COVID-19 Comes 40 Years After AIDS - Any Lesson?

text: The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has hit health-care systems and societies in an unprecedented manner. In 1981, the first cases of AIDS were reported and wide diagnostic testing helped to characterize high-risk groups and the global burden of the epidemic. With Coronavirus Disease (COVID)-19, everything has happened too fast and both cases and fatalities are huge but still uncertain in most places. Diagnostic testing of active and past SARS-CoV-2 infections needs to expand rapidly, ideally using rapid tests. COVID-19 deaths are highly concentrated in the elderly population, with a large proportion of fatalities being "with" rather than "by" SARS-CoV-2 infection. They are often the result of inadequate health care due to overwhelming demands. To date, there is no specific therapy for SARS-CoV-2 infection. Several antivirals are being tested clinically, including remdesivir, at this time the most promising. For others such as lopinavir/ritonavir, neither significant virological nor clinical benefit has been shown. Given the characteristic pulmonary cytokine storm underlying the pathogenic mechanism of severe COVID-19 pneumonia and acute respiratory distress, antiinflammatory agents are being investigated. The benefit of orticosteroids, hydroxychloroquine, etc., is limited. Monoclonal antibodies targeting different pro-inflammatory cytokines, such as tocilizumab, an anti-interleukin 6 agent, are being tried with encouraging results. Ultimately a protective vaccine will be the best response for controlling the COVID-19 pandemic.


title: Functional respiratory imaging, regional strain, and expiratory time constants at three levels of positive end expiratory pressure in an ex vivo pig model.

text: Heterogeneity in regional end expiratory lung volume (EELV) may lead to variations in regional strain (ε). High ε levels have been associated with ventilator-associated lung injury (VALI). While both whole lung and regional EELV may be affected by changes in positive end-expiratory pressure (PEEP), regional variations are not revealed by conventional respiratory system measurements. Differential rates of deflation of adjacent lung units due to regional variation in expiratory time constants (τE) may create localized regions of ε that are significantly greater than implied by whole lung measures. We used functional respiratory imaging (FRI) in an ex vivo porcine lung model to: (i) demonstrate that computed tomography (CT)-based imaging studies can be used to assess global and regional values of ε and τE and, (ii) demonstrate that the manipulation of PEEP will cause measurable changes in total and regional ε and τE values. Our study provides three insights into lung mechanics. First, image-based measurements reveal egional variation that cannot be detected by traditional methods such as spirometry. Second, the manipulation of PEEP causes global and regional changes in R, E, ε and τE values. Finally, regional ε and τE were correlated in several lobes, suggesting the possibility that regional τE could be used as a surrogate marker for regional ε.


title: Elimination of phosphorylation sites of Semliki Forest virus replicase protein nsP3.

text: nsP3 is one of the four RNA replicase subunits encoded by alphaviruses. The specific essential functions of nsP3 remain unknown, but it is known to be phosphorylated on serine and threonine residues. Here we have completed mapping of the individual phosphorylation sites on Semliki Forest virus nsP3 (482 amino acids) by point mutational analysis of threonine residues. This showed that threonines 344 and 345 represented the major threonine phosphorylation sites in nsP3. Experiments with deletion variants suggested that nsP3 itself had no kinase activity; instead, it was likely to be phosphorylated by multiple cellular kinases. Phosphorylation was not necessary for the peripheral membrane association of nsP3, which was mediated by the N-terminal region preceding the phosphorylation sites. Two deletion variants of nsP3 with either reduced or undetectable phosphorylation were studied in the context of virus infection. Cells infected with mutant viruses produced close to wild type levels of infectious virions; however, the rate of viral RNA synthesis was significantly reduced in the mutants. A virus totally defective in nsP3 phosphorylation and exhibiting a decreased rate of RNA synthesis also exhibited greatly reduced pathogenicity in mice.


title: Colostral and milk antibody titers in cows vaccinated with a modified live-rotavirus-coronavirus vaccine.

text: Colostral and milk whey rotavirus (RV) and coronavirus (CV) antibody titers stimulated in 15 beef heifers by vaccination with a modified live-RV-CV vaccine were compared with titers in 15 nonvaccinated heifers. Geometric mean antibody titers to RV in colostral and 3-day milk whey from vaccinated heifers were 2,807 and 92, respectively, and in control heifers were 1,613 and 71, respectively. Geometric mean antibody titers to CV in colostral and 3-day milk whey of vaccinated heifers were 877 and 13, respectively, compared with titers were 877 and 13, respectively, compared with titers in nonvaccinated heifers of 731 and 7, respectively. Differences in antibody titers between vaccinated and nonvaccinated heifers were not significantly different.


title: [Viruses as causal agents of gastroenteritis in infants and young children (author's transl)].

text: Electron microscopic examination of stool samples from 195 infants and young children with acute gastroenteritis revealed rota virus in 31 samples (= 16%), corona virus in 44 samples (= 23%) and adenovirus in 13 samples (= 7%). Viruses were excreted for between 4 and 8 days in patients infected with rota virus and corona virus respectively. An outbreak of acute gastroenteritis in a premature baby unit with 24 cases is reported: corona virus was found in the stools of 15 infants. It is suggested that corona virus as well as rota virus may cause gastroenteritis in infants and small children.


title: Role of rhinovirus in hospitalized infants with respiratory tract infections in Spain.

text: BACKGROUND Rhinovirus is a recognized cause of common cold, proven to cause asthma exacerbations in children. In Spain, no description exists, as yet, as to the degree of burden rhinovirus infections represent among hospitalized infants. Our aim was to describe rhinovirus infections in hospitalized children, under 2 years of age, and to compare these with patients infected with respiratory syncytial virus (RSV). PATIENTS AND METHODS The prospective study was performed between September 2003 and July 2005, in children <2 years of age, admitted at the Severo Ochoa Hospital (Leganés, Madrid) with fever or respiratory tract infection and with positive rhinovirus detection in the nasopharyngeal aspirate samples. Virologic diagnosis was made by multiplex reverse transcription-polymerase chain reaction and for some virus by direct immunofluorescent assay in nasopharyngeal samples. Demographic and clinical data of those patients with rhinovirus infection were described and compared with a group of 86 patients, infected only with RSV, randomly selected from the same population. RESULTS We detected 85 children admitted to hospital with rhinovirus infection. Rhinovirus was the cause of 25% of all admissions, among the total of 340 under 2-year olds diagnosed with fever or respiratory tract infection. Rhinovirus was the second viral agent identified, after RSV. Clinical diagnosis was recurrent wheezing in 48.2%; bronchiolitis in 36.5%; and pneumonia in 3.5%. Fever was present in 60% of the patients. Radiologic infiltrates were found in 22.4% of the children. In 50.6% of the infants, oxygen saturation under 95% was detected, at the time of admission. Hypoxia was present in RSV-infected children more frequently (P = 0.005). Also, in this group, final diagnosis was, most frequently, bronchiolitis (P = 0.0001), and rhinovirus-infected patients were most frequently males (P = 0.004). CONCLUSIONS Rhinovirus was detected in hospitalized infants with respiratory tract disease and was the second most common virus after RSV. In our experience, it was the second etiologic agent associated with recurrent wheezing in hospitalized children, under the age of 2 years.


title: [Anti-virus research of triterpenoids in licorice].

text: Licorice is a leguminous plant of glycyrrhiza. It is a traditional Chinese herbal medicine. Triterpenoid is one of the mainly active components of licorice. In recent years, the broad-spectrum antiviral activity of many triterpenoids in licorice was confirmed, and these findings have become a hot spot of antiviral immunity. The triterpenoids of licorice has the potential to become a novel broad-spectrum antiviral medicine and will be widely used in the clinical treatment. This review provided a summary of the recent anti-virus research progress on several triterpenoids in licorice, such as glycyrrhizic acid, glycyrrhizin, glycyrrhetinic acid and its derivatives. The antiviral roles of triterpenoids in licorice against herpes virus, HIV, hepatitis virus, SARS coronavirus and influenza virus were briefly summarized.


title: Stress-induced mutagenesis in bacteria.

text: The evolutionary significance of stress-induced mutagenesis was evaluated by studying mutagenesis in aging colonies (MAC) of Escherichia coli natural isolates. A large fraction of isolates exhibited a strong MAC, and the high MAC variability reflected the diversity of selective pressures in ecological niches. MAC depends on starvation, oxygen, and RpoS and adenosine 3',5'-monophosphate regulons; thus it may be a by-product of genetic strategies for improving survival under stress. MAC could also be selected through beneficial mutations that it generates, as shown by computer modeling and the patterns of stress-inducible and constitutive mutagenesis. We suggest that irrespective of the causes of their emergence, stress-induced mutations participate in adaptive evolution.


title: "Exposing the cracks": Severe Acute Respiratory Syndrome and the Hong Kong health system.

text: PURPOSE Hong Kong was particularly affected by the 2003 outbreak of Severe Acute Respiratory Syndrome (SARS). During the epidemic, it seemed as if the Hong Kong government and health system were barely coping, leading to calls of mismanagement and governance incapacity. In the wake of the SARS outbreak, two inquiries were conducted. The purpose of this article is to review the Hong Kong's response to SARS from the perspective of two inquiries. DESIGN/METHODOLOGY/APPROACH An historical analysis of the institutional arrangements for health care delivery in Hong Kong is undertaken, followed by a chronology of developments in the SARS outbreak. The article then reviews outbreak management and the findings of the two inquiries. Finally, it considers whether the Hong Kong health system can be reformed to manage any future infectious disease epidemic better. FINDINGS Both leadership and coherency were lacking in Hong Kong's response to SARS. These are age-old problems in the Hong Kong health sector. The prospects for mending the health system appear limited, given that leadership and coherency have been consistently absent features of post-1997 governance in Hong Kong. RESEARCH LIMITATIONS/IMPLICATIONS This article reviews events in the immediate period following the SARS outbreak. A future follow-up study of the Hong Kong government and health system's capacity to respond to infectious disease outbreaks would be useful. PRACTICAL IMPLICATIONS This article provides a review that will be useful to policymakers and researchers. ORIGINALITY/VALUE No other article reviews the Hong Kong health system's SARS response.