title: Reduced incorporation of SARS-CoV spike protein into viral particles due to amino acid substitutions within the receptor binding domain.

text: Cell clone #21 is a long-term producer of the infectious SARS-coronavirus, although the incorporation rate of spike (S) protein into virions is significantly lower. Sequencing analysis of the viral structural proteins revealed four and one amino acid substitutions in the S and membrane (M) proteins, respectively. We demonstrated, using a viral-like particle formation system, that the S mutations were involved in the lower incorporation of the S protein into virions, although the M mutation that disrupts the glycosylation was not present in this phenotype. Further mutational experiments identified two substitutions, Y442C and L472F, within the receptor binding domain that could be critical for the reduced S incorporation, as well as reduced binding affinity between the S protein and ACE2 receptor. Thus, these two amino acid substitutions might lead to a conformational change in the S protein, resulting in reduced incorporation into viral particles.


title: Prolonged motor block following epidural anaesthesia: A proposed pathway for investigation and management to facilitate rapid MRI scanning to exclude vertebral canal haematoma.

text: Vertebral canal haematoma following epidural anaesthesia is a rare but potentially devastating occurence, which can lead to permanent neurological damage. Early clinical identification and diagnosis using MRI imaging with surgical decompression of the haematoma can lead to full recovery (Kebaish 2004). However surgical patients often have metal clips or staples, which are left inside the body, some of which are not MRI compatible. Currently there is no process to document which surgical clips have been used and their MRI compatibility in an accessible format. This can lead to a delay in performing MRI imaging. We propose a clinical pathway to expedite diagnosis of vertebral canal haematoma to incorporate a system to allow rapid identification of MRI compatibility of surgical clips used during surgery.


title: Catheter Related Right Atrial Thrombus in Patients on Maintenance Hemodialysis: Results of a Single Centre Retrospective Study from a Tertiary Care Hospital.

text: Objective To study the magnitude of the complication of catheter associated right atrial thrombus (CRAT) in patients with tunnelled central venous hemodialysis catheters (THC) for maintenance hemodialysis (MHD). Material and methods A retrospective study was conducted among patients with end stage kidney disease (ESKD) with THC for MHD who had undergone screening for CRAT with a 2D-echo (2DE) just before removal of the THC. The occurrence of CRAT and other clinical parameters were documented in these patients. Results A total of 28 patients (mean [SD] age 51 [15.2] years; females 17 [60.7%]) were included in the study. CRAT was observed in 5 (17.9%) patients. There was no difference in mean age in patients with or without thrombus (48±13.02 vs 51.61 ± 15.78 years; p = 0.61). History of diabetes and hypertension was present in 2 and all 5 patients respectively. There was no significant difference in the period the THC was in place in patients with or without CRAT (13±7.8 months vs 10.57±5.66 months; p = 0.54). There was no association between catheter related blood stream infection (CRBSI) and CRAT (p= 0.29). Conclusion The incidence of CRAT in patients with THC for MHD was 17.9%. Patients with THC for MHD should be examined for presence of CRAT before removal of THC to prevent fatal pulmonary thromboembolism.


title: Push-Ups vs. Bench Press Differences in Repetitions and Muscle Activation between Sexes.

text: Push-ups are an ubiquitous resistance training exercise. While exhibiting a relatively similar upper body motion to the bench press, there are substantial differences in repetitions when employing similar relative loads. The objective was to examine sex-related differences in repetitions and muscle activation associated with push-ups and bench press exercises. Twenty resistance-trained participants (10 men [22 ± 6.1 years] and 10 [24 ± 5.7 years] women) performed maximum push-up and bench press repetitions with loads relative to the body mass during a push-up. Electromyographic (EMG) electrodes were positioned on the middle and anterior deltoids, triceps and biceps brachii, and pectoralis major muscles and their relative (normalized to a maximum voluntary contraction) activity was compared between the two exercises performed to task failure. Both females (3.5 ± 3.9 vs.15.5 ± 8.0 repetitions; p = 0.0008) and males (12.0 ± 6.3 vs. 25.6 ± 5.2 repetitions; p < 0.0001) performed 77.4% and 53.1% less bench press than push-up repetitions respectively. Males significantly exceeded females with both push-ups (p = 0.01) and bench press (p = 0.004) repetitions. Significant linear regression equations were found for females (r2 = 0.55; p = 0.03), and males (r2 = 0.66; p < 0.0001) indicating that bench press repetitions increased 0.36 and 0.97 for each push-up repetition for females and males respectively. Triceps (p = 0.002) and biceps brachii (p = 0.03) EMG mean amplitude was significantly lower during the push-up concentric phase, while the anterior deltoid (p = 0.03) exhibited less activity during the bench press eccentric phase. The sex disparity in repetitions during these exercises indicates that a push-up provides a greater challenge for women than men and regression equations may be helpful for both sexes when formulating training programs.


title: Synthetic peptides outside the spike protein heptad repeat regions as potent inhibitors of SARS-associated coronavirus.

text: A novel severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) has been identified as the aetiological agent of SARS. We previously isolated and characterized SARS-CoV and SARS-CoV-like viruses from human and animals, respectively, suggesting that SARS could be transmitted from wild/farmed animals to humans. Comparison of the viral genomes indicated that sequence variation between animal and human isolates existed mainly in the spike (S) gene. We hypothesized that these variations may underlie a change of binding specificity of the S protein to the host cells, permitting viral transmission from animals to humans. Here we report that four 20-mer synthetic peptides (S protein fragments), designed to span these sequence variation hotspots, exhibited significant antiviral activities in a cell line. SARS-CoV infectivity was reduced over 10 000-fold through pre-incubation with two of these peptides, while it was completely inhibited in the presence of three peptides. Molecular modelling of the SARS-CoV peplomer suggests that three of these antiviral peptides map to the interfaces between the three monomers of the trimeric peplomer rather than the heptad repeat region from which short peptides are known to inhibit viral entry. Our results revealed novel regions in the spike protein that can be targeted to inhibit viral infection. The peptides identified in this study could be further developed into antiviral drugs.


title: [Side effects of glucocorticosteroids in the management of 1291 patients of SARS].

text: OBJECTIVE To analysis the relationship between glucocorticosteroids (GCS) usage and side effects in the treatment of severe acute respiratory syndrome (SARS). METHODS All clinical records of probable SARS patients in Beijing were collected and input into an Epi6 database, in which 1 291 patients had entire information and met the clinical criteria of SARS. The usage of GCS and GCS associated side effects were analyzed retrospectively. RESULTS Patients accepted GCS therapy were 83.96% (n=1 084), whereas 16.04%(n=207) did not take GCS. The average dosage of GCS was 160 mg/d in the first week, and then reduced to 80 mg/d and 40 mg/d in the second and the third weeks, respectively. Initial blood glucose, systolic pressure (SBP), and diastolic pressure (DBP) were no significant difference between GCS group and non-GCS group. The highest blood glucose during the treatment in GCS group was markedly higher than that in non-GCS group [(8.68+/-4.80) mmol/L vs (6.39+/-3.71) mmol/L, P<0.05)]. The highest blood glucose and average blood glucose after initiation of GCS therapy were elevated in GCS group. The levels of blood glucose were correlated with the initial, maximum, and cumulative GCS dosages. Average blood glucose was increased markedly in groups with MP(Initial) > or =80 mg/d (Methyprednisonlone), MP(Maximal) > or =160 mg/d, or MP(Cumulative) > or = 3000 mg. The blood glucose grew up significantly in the first and the second weeks in the treatment with GCS, and then returned to normal level gradually. Hyperglycemia duration in the group with MP(Cumulative) > or =3000 mg persisted longer than that in the other groups (P< 0.05). The lowest serum potassium during the treatment and the duration of hypokalemia in GCS group were significantly different from that in non-GCS group [(3.66+/-0.50) mmol/L vs (4.01+/-0.51) mmol/L, P< 0.001 ;1(1, 75) days vs 1(1, 9) days, P<0.05, respectively]. Average serum potassium and the duration of hypokalemia were related to the dosages of GCS. Serum potassium reached its nadir in the first week of GCS treatment and then grew up in the second week. In groups with MP(Initial) > or =320 mg/d, MP(Maximal) > or =320 mg/d, and MP(Cumulative) > or =3000 mg, the level of serum potassium was lower and the duration of hypokalemia was longer than that in other groups. They began to returned to normal level in the third week. Administration of GCS prolonged the time of hypocalcemia[19 (1, 74) days in GCS group vs 8 (1, 32) days in non-GCS group, P< 0.05]. The duration of hypocalcemia was prolonged according to the increasing of the maximal or the cumulative dosage of GCS. However, the duration of hypocalcemia in group with MP(Cumulative) <999 mg was similar to that in non-GCS group (P > 0.05). After GCS administration, SBP and DBP were increased gradually, and reached their peaks in the fourth week [SBP(117.2+/-14.0) mm Hg and DBP (72.5+/-9.1) mm Hg vs SBP (120.0+/-12.5) mm Hg and DBP (74.5+/-8.7) mm Hg, P< 0.05, 1 mm Hg=0.133 kPa]. CONCLUSION Hyperglycemia and hypokalemia are correlated with GCS dosage and duration. Administration with GCS influences SBP, DBP, and duration of hypocalcemia. Appropriate low dosage of GCS (MP(Initial) and MP(Maximal) < 159 mg/d, MP(Cumulative)< 2999 mg) causes few changes of blood glucose, serum potassium, and blood calcium. It is important to monitor laboratory findings during the treatment with GCS.


title: Single-incision laparoscopic distal pancreatectomy.

text: Single-incision laparoscopic surgery (SILS) is a novel area of minimally invasive surgery, and rapidly evolving in the field of abdominal surgery. However, SILS is not a common procedure for pancreatic surgery. We describe our first experience with a SILS approach in a patient with a metastatic pancreatic tumor. We performed a SILS distal pancreatectomy (SILS-DP) using 3 trocars placed through a single port at an umbilical incision. In our case, the most important surgical technique was the stomach-hanging method to obtain a favorable laparoscopic view of the pancreas. Although SILS-DP is a safe and feasible procedure, further studies are required to determine the advantages of this procedure in comparison with the standard laparoscopic method.


Shaping the future of global health.


Q&A with Albert S. Woo, MD Forging a New Frontier in 3D Printing during COVID-19 Pandemic.


Pathogenesis of rhinovirus common colds in human voluteers.


title: Outbreak of febrile respiratory illness caused by adenovirus at a South Korean military training facility: clinical and radiological characteristics of adenovirus pneumonia.

text: Adenovirus often causes respiratory disease outbreaks in military training soldiers. Compared with adenovirus pneumonia in young military adults, symptoms arising from febrile respiratory illness (FRI) caused by adenovirus have not been previously evaluated in Korean soldiers. We conducted an event-based outbreak investigation involving 712 male soldiers aged 19 to 21 years from March 14 to 30, 2012 to evaluate the epidemiological and clinical characteristics of patients with pneumonia and FRI caused by adenovirus. We described the laboratory and radiological characteristics of patients with adenovirus pneumonia. Among these, 407 cases of FRI and 15 cases of pneumonia were identified through active surveillance (attack rate of FRI, 57.16%; attack rate of pneumonia, 2.11%). Fire training and march training may present environmental risk factors for adenovirus-associated outbreaks. Most symptoms were mild. The most frequent symptom in patients with pneumonia and FRI was cough. Patients with pneumonia were associated with an increased incidence of dizziness (crude odds ratio [cOR], 9.65; 95% confidence interval [CI], 2.38-37.15) and a decreased incidence of rhinorrhea (cOR, 0.15; 95% CI, 0.04-0.53) during adenovirus-associated outbreaks. Differential leukocyte count revealed high monocytes, low lymphocytes, and low eosinophils, and chest computed tomography revealed a consolidation pattern and right lobar pneumonia. These findings warrant a high level of suspicion for adenovirus pneumonia.


Polypeptide structure of bovine enteritic coronavirus: comparison between a wild strain purified from feces and a HRT 18 cell adapted strain.


title: A new lead for nonpeptidic active-site-directed inhibitors of the severe acute respiratory syndrome coronavirus main protease discovered by a combination of screening and docking methods.

text: The coronavirus main protease, M(pro), is considered to be a major target for drugs suitable for combating coronavirus infections including severe acute respiratory syndrome (SARS). An HPLC-based screening of electrophilic compounds that was performed to identify potential M(pro) inhibitors revealed etacrynic acid tert-butylamide (6a) as an effective nonpeptidic inhibitor. Docking studies suggested a binding mode in which the phenyl ring acts as a spacer bridging the inhibitor's activated double bond and its hydrophobic tert-butyl moiety. The latter is supposed to fit into the S4 pocket of the target protease. Furthermore, these studies revealed etacrynic acid amide (6b) as a promising lead for nonpeptidic active-site-directed M(pro) inhibitors. In a fluorimetric enzyme assay using a novel fluorescence resonance energy transfer (FRET) pair labeled substrate, compound 6b showed a K(i) value of 35.3 muM. Since the novel lead compound does not target the S1', S1, and S2 subsites of the enzyme's substrate-binding pockets, there is room for improvement that underlines the lead character of compound 6b.


title: Bioinformatics research on the SARS coronavirus (SARS_CoV) in China.

text: Severe acute respiratory syndrome (SARS) first appeared in 2002 in China, which fastly affected about 8000 patients over 29 countries and caused 774 fatalities. As its pathogen was identified as a new kind of coronavirus (SARS_CoV), its genome was quickly sequenced on several isolates. Studies on its functional genomics were performed by combinatorial application of all the available bioinformatics tools and the development of new programs. In this way, it was found that the four proteins were absolutely responsible for nosogenesis of SARS, i.e. spike (S) protein; small envelop (E) protein; membrane (M) protein; and nucleocaspid (N) protein. Molecular evolution studies have revealed that SARS must be originated from wild animals, and it was demonstrated that the major genetic variations in some critical genes, particularly the Spike gene, was essential for the transition from animal-to-human transmission to human-to-human transmission. Theoretical models, either Logistic model or SIR model, were developed to describe the transmission of SARS. The recorded difference of SARS spreading in Beijing and Hong Kong was also reasonably analyzed according to these models. The whole process of fruitful bioinformatics studies, along with other related scientific investigations have set up an unprecedented paradigm for human of how to battle against sudden-breaking and catastrophic epidemics.


title: Preparing for avian influenza.

text: PURPOSE OF REVIEW Pandemic influenza is inevitable, will have significant impact on the population and will challenge healthcare delivery. Planning for an influenza pandemic will improve the ability of healthcare facilities to respond successfully. RECENT FINDINGS Planning activities involve determining the role of the facility during an influenza pandemic, estimating resources necessary to fulfil this role and developing the infrastructure, material and human resources surge capacity necessary to meet the estimated needs. Plans to perform surveillance for potentially contagious individuals, handle clinical specimens, increase social distancing during a pandemic and the development of personal preparedness plans may be beneficial. SUMMARY The goal of this review is to provide a framework for healthcare facility pandemic influenza planning.


title: Colocalization and interaction of the porcine arterivirus nucleocapsid protein with the small nucleolar RNA-associated protein fibrillarin.

text: Porcine reproductive and respiratory syndrome virus (PRRSV) replicates in the cytoplasm of infected cells, but its nucleocapsid (N) protein localizes specifically to the nucleus and nucleolus. The mechanism of nuclear translocation and whether N associates with particular nucleolar components are unknown. In the present study, we show by confocal microscopy that the PRRSV N protein colocalizes with the small nucleolar RNA (snoRNA)-associated protein fibrillarin. Direct and specific interaction of N with fibrillarin was demonstrated in vivo by the mammalian two-hybrid assay in cells cotransfected with the N and fibrillarin genes and in vitro by the glutathione S-transferase pull-down assay using the expressed fibrillarin protein. Using a series of deletion mutants, the interactive domain of N with fibrillarin was mapped to a region of amino acids 30 to 37. For fibrillarin, the first 80 amino acids, which contain the glycine-arginine-rich region (the GAR domain), was determined to be the domain interactive with N. The N protein was able to bind to the full-length genomic RNA of PRRSV, and the RNA binding domain was identified as the region overlapping with the nuclear localization signal situated at positions 41 to 47. These results suggest that the N protein nuclear transport may be controlled by the binding of RNA to N. The PRRSV N protein was also able to bind to both 28S and 18S ribosomal RNAs. The protein-protein interaction between N and fibrillarin was RNA dependent but independent of N protein phosphorylation. Taken together, our studies demonstrate a specific interaction of the PRRSV nucleocapsid protein with the host cell protein fibrillarin in the nucleolus, and they imply a potential linkage of viral strategies for the modulation of host cell functions, possibly through rRNA precursor processing and ribosome biogenesis.


The role of public health and occupational physicians in the new millennium.


title: Feline immunodeficiency virus infection in a population of pet cats from southeastern Florida.

text: Blood samples from 95 randomly selected pet cats that were brought to veterinarians in southeastern Florida were tested for antibodies to feline immunodeficiency virus (FIV). Virus-specific antibodies (indicative of virus infection) were found in 8 of the 95 (8.4%) cats tested. All of the virus-infected cats were males (statistically significant, P less than or equal to 0.016) and were at least 1 year of age. The 3 most severely ill cats infected with FIV were also infected with feline leukemia virus.


title: Outbreak of necrotizing enterocolitis associated with Enterobacter sakazakii in powdered milk formula.

text: We describe an outbreak of necrotizing enterocolitis (NEC) that occurred in the neonatal intensive care unit of our hospital. A total of 12 neonates developed NEC in June-July 1998. For two of them, twin brothers, the NEC turned out to be fatal. Enterobacter sakazakii, a known contaminant of powdered milk formula, was isolated from a stomach aspirate, anal swab, and/or blood sample for 6 of the 12 neonates. A review of feeding procedures revealed that 10 of the 12 patients were fed orally with the same brand of powdered milk formula. E. sakazakii was isolated from the implicated prepared formula milk as well as from several unopened cans of a single batch. Molecular typing by arbitrarily primed PCR (AP-PCR) confirmed, although partially, strain similarity between milk and patient isolates. No further cases of NEC were observed after the use of the contaminated milk formula was stopped. With this outbreak we show that intrinsic microbiological contamination of powdered milk formula can be a possible contributive factor in the development of NEC, a condition encountered almost exclusively in formula-fed premature infants. The use of sterilized liquid milk formula in neonatal care could prevent problems with intrinsic and extrinsic contamination of powdered milk formula.


title: Human metapneumovirus: a ubiquitous and long-standing respiratory pathogen.

text: BACKGROUND Human metapneumovirus (hMPV) is a recently described human pathogen first identified in respiratory specimens of young children suffering from respiratory syndromes ranging from mild to severe. METHODS AND RESULTS Virological studies have reported the presence of hMPV infections in many countries from all continents. Seroprevalence studies have indicated that the virus has been circulating in humans for more than 50 years and that it infects virtually all children by the ages of 5-10 years. In young children, hMPV has been mainly associated with bronchiolitis but also with pneumonitis, otitis media and acute exacerbation of asthma. The contribution of hMPV in respiratory syndromes of adults has been studied considerably less; initial studies have indicated a role for this pathogen in flu-like syndromes and in significant percentages of chronic obstructive pulmonary disease exacerbations and cases of community-acquired pneumonia during the winter-spring period. Both primate and rodent experimental models have been used to characterize the pathogenesis of this respiratory virus. In some of these models, intranasal hMPV inoculation has elicited not only important viral replication but also significant pulmonary inflammation and clinical disease. Recently a few groups have developed reverse genetic systems for hMPV, allowing a better understanding of viral pathogenesis and generation of attenuated viral strains for immunization. CONCLUSIONS Recent studies on hMPV have provided a better understanding of the epidemiology and pathogenesis associated with this viral infection, and have enhanced the prospect of developing efficient therapeutic agents and vaccine candidates.


title: [Evaluation of tracheal intubation in critical SARS patients].

text: OBJECTIVE To analyze tracheal intubation and respiratory treatment in the critical severe acute respiratory syndrome (SARS) patients. METHODS Review and analyze tracheal intubation and respiratory treatment in critical SARS patients in intensive care unit (ICU). RESULTS Three of thirteen patients had been intubated or received tracheotomy before they entered into ICU, the other patients received treatment of nasal cannula or oxygen mask. With the development of the disease, two patients had been intubated because of respiratory failure or tracheotomy. Tracheal intubation was twice made in two patients in order to replace tracheal tubes. CONCLUSIONS The patient should be intubated or received tracheotomy if non-invasive respiratory support has no effect. Standard protection could protect medical staff from infection under tracheal intubation.


title: Confronting zoonoses through closer collaboration between medicine and veterinary medicine (as 'one medicine').

text: In the 19th century, the concept of 'one medicine' was embraced by leaders in the medical and veterinary medical communities. In the 20th century, collaborative efforts between medicine and veterinary medicine diminished considerably. While there have been some notable exceptions, such as Calvin W. Schwabe's proposal for unifying human and veterinary medicine and joint efforts by the Food and Agriculture Organization and World Health Organization to control zoonotic diseases, 'one medicine' has languished in the modern milieu of clinical care, public health, and biomedical research. Risks of zoonotic disease transmission are rarely discussed in clinical care which is of particular concern if humans and/or animals are immunosuppressed. Physicians and veterinarians should advise their patients and pet-owning clients that some animals should not be pets. The risk of zoonotic disease acquisition can be considerable in the occupational setting. Collaborative efforts in biomedical research could do much to improve human and animal health. As the threat of zoonotic diseases continues to increase in the 21st century, medicine and veterinary medicine must revive 'one medicine' in order to adequately address these challenges. 'One medicine' revival strategies must involve medical and veterinary medical education, clinical care, public health and biomedical research.


title: Laryngeal Mask Airway and Valsalva Maneuver During Ophthalmic Surgery: A Case Report.

text: The use of a supraglottic airway, also called laryngeal mask airway (LMA), is gaining in popularity for delivery of general anesthesia in certain situations and is shown to be safe. Use of infraglottic airways (tracheal tubes) allows for positive pressure ventilation and maneuvers, such as holding continuous positive pressure in the airway to check for pleural and dural leaks. We describe the successful use of the LMA (Teleflex Inc) for general anesthesia, in which continuous positive airway pressure of 15 cm H2O held for 20 seconds allowed the neurosurgeon to identify the site of a cerebrospinal fluid (CSF) leak and successfully repair the leak. A well-seated LMA can be successfully used to produce continuous positive pressure in the airway not exceeding 20 cm H2O for 20 seconds to facilitate surgical identification of CSF leaks.


title: Neutralizing antibodies against feline herpesvirus type 1 in captive wild felids of Brazil.

text: Feline herpesvirus type 1 infection affects domestic cats, causing mainly upper respiratory tract diseases. Although this infection has been described in captive and free-ranging wild felids from Europe, Asia, North America, and Africa, no information is available on its occurrence among wild felids of Brazil. In this study, 250 serum samples of six species of Brazilian captive wild felids (Leopardus tigrinus, Leopardus wiedii, Herpailurus yaguarondi, Puma concolor, Leopardus pardalis, and Panthera onca) were examined for neutralizing antibodies to feline herpesvirus type 1. Positive sera were found in 72% of L. tigrinus samples, 15% of L. wiedii, 6% of L. pardalis, 8% of H. yaguarondi, 18% of P. concolor, and 14% of P. onca. The relatively low percentages of seropositivity and low antibody titers found among the last five species suggest that feline herpesvirus type 1 does not circulate extensively among these animals. Nevertheless, quarantine, serologic screening, and vaccination of newly introduced felids is recommended in zoos in order to prevent virus transmission and outbreaks of the disease among wild felids kept in captivity.


title: Thick lung wedge resection for acute life-threatening massive hemoptysis due to aortobronchial fistula.

text: Massive hemoptysis from an aortobronchial fistula due to thoracic aortic dissection is an extremely rare symptom, but is a potentially life-threatening condition. We report a case of acute massive hemoptysis due to aortobronchial fistula that was successfully controlled by a simple and rapid thick wedge resection of the lung with hematoma by using the black cartilage stapler. A 65-year-old man was admitted to our hospital with acute massive hemoptysis. After tracheal intubation, chest computed tomography revealed hematoma in the left lung and ruptured aortic dissection from the distal arch to the descending aorta. He was diagnosed with aortobronchial fistula and underwent an emergency surgery on the same day. We performed posterolateral thoracotomy. A dissecting aortic aneurysm (diameter, ~80 mm) with adhesion of the left upper lobe and the superior segment of the lower lobe was found. The lung parenchyma expanded with the hematoma. We stapled the upper and lower lobes by using the black cartridge stapler along the aortopulmonary window. Massive hemoptysis disappeared, and the complete aortic dissection appeared. Aortic dissection with adherent lung was excised, and graft replacement of the distal arch and descending thoracic aorta was performed. Proximal lung wedge resection using black cartridge stapler is a simple and quick method to control massive hemoptysis from aortic dissection; hence, this procedure is an effective option to control massive hemoptysis due to aortobronchial fistula. This technique could rapidly stop massive hemoptysis and prevent dissection of the adherent lung tissue and intra-thoracic bleeding.


title: Multiple vitamin K-dependent coagulation zymogens promote adenovirus-mediated gene delivery to hepatocytes.

text: Upon local delivery, adenovirus (Ad) serotype 5 viruses use the coxsackie and Ad receptor (CAR) for cell binding and alpha(v) integrins for internalization. When administered systemically, however, their role in liver tropism is limited because CAR-permissive and mutated viruses show similar biodistribution, a finding recently attributed to blood coagulation factor (F) IX or complement protein C4BP binding to the adenovirus fiber and "bridging" to either low-density lipoprotein receptor-related protein or heparan sulfate proteoglycans. Here, we show that hepatocyte transduction in vitro can be enhanced by the vitamin K-dependent factors FX, protein C, and FVII in addition to FIX but not by prothrombin (FII), FXI, and FXII. This phenomenon was not dependent on proteolytic activation or cell signaling activity and for FX was mediated by direct virus-factor binding. Human FX substantially enhanced hepatocyte transduction by CAR-permissive and mutated viruses in an ex vivo liver perfusion model. In vivo, global down-regulation of vitamin K-dependent zymogens by warfarin significantly diminished liver uptake of CAR-deleted Ads; however, this phenomenon was fully rescued by acute infusion of human FX. Our results indicate a common and pivotal role for distinct vitamin K-dependent coagulation factors in mediating hepatocyte transduction by adenoviruses in vitro and in vivo.


title: Design of multichannel functional near-infrared spectroscopy system with application to propofol and sevoflurane anesthesia monitoring.

text: Monitoring the changes of cerebral hemodynamics and the state of consciousness during general anesthesia (GA) is clinically important. There is a great need for developing advanced detectors to investigate the physiological processes of the brain during GA. We developed a multichanneled, functional near-infrared spectroscopy (fNIRS) system device and applied it to GA operation monitoring. The cerebral hemodynamic data from the forehead of 11 patients undergoing propofol and sevoflurane anesthesia were analyzed. The concentration changes of oxygenated hemoglobin, deoxygenated hemoglobin, total hemoglobin, and cerebral tissue heart rate were determined from the raw optical information based on the discrete stationary wavelet transform. This custom-made device provides an easy-to-build solution for continuous wave-fNIRS system, with customized specifications. The developed device has a potential value in cerebral monitoring in clinical settings.


title: Understanding helicases as a means of virus control.

text: Helicases are promising antiviral drug targets because their enzymatic activities are essential for viral genome replication, transcription, and translation. Numerous potent inhibitors of helicases encoded by herpes simplex virus, severe acute respiratory syndrome coronavirus, hepatitis C virus, Japanese encephalitis virus, West Nile virus, and human papillomavirus have been recently reported in the scientific literature. Some inhibitors have also been shown to decrease viral replication in cell culture and animal models. This review discusses recent progress in understanding the structure and function of viral helicases to help clarify how these potential antiviral compounds function and to facilitate the design of better inhibitors. The above helicases and all related viral proteins are classified here based on their evolutionary and functional similarities, and the key mechanistic features of each group are noted. All helicases share a common motor function fueled by ATP hydrolysis, but differ in exactly how the motor moves the protein and its cargo on a nucleic acid chain. The helicase inhibitors discussed here influence rates of helicase-catalyzed DNA (or RNA) unwinding by preventing ATP hydrolysis, nucleic acid binding, nucleic acid release, or by disrupting the interaction of a helicase with a required cofactor.


title: Management of Fever in Infants and Young Children.

text: Despite dramatic reductions in the rates of bacteremia and meningitis since the 1980s, febrile illness in children younger than 36 months continues to be a concern with potentially serious consequences. Factors that suggest serious infection include age younger than one month, poor arousability, petechial rash, delayed capillary refill, increased respiratory effort, and overall physician assessment. Urinary tract infections are the most common serious bacterial infection in children younger than three years, so evaluation for such infections should be performed in those with unexplained fever. Abnormal white blood cell counts have poor sensitivity for invasive bacterial infections; procalcitonin and C-reactive protein levels, when available, are more informative. Chest radiography is rarely recommended for children older than 28 days in the absence of localizing signs. Lumbar puncture is not recommended for children older than three months without localizing signs; it may also be considered for those from one to three months of age with abnormal laboratory test results. Protocols such as Step-by-Step, Laboratory Score, or the Rochester algorithms may be helpful in identifying low-risk patients. Rapid influenza testing and tests for coronavirus disease 2019 (COVID-19) may be of value when those diseases are circulating. When empiric treatment is appropriate, suggested antibiotics include ceftriaxone or cefotaxime for infants one to three months of age and ampicillin with gentamicin or with cefotaxime for neonates. For children three months to three years of age, azithromycin or amoxicillin is recommended if pneumonia is suspected; for urinary infections, suggested antibiotics are cefixime, amoxicillin/clavulanate, or trimethoprim/sulfamethoxazole. Choice of antibiotics should reflect local patterns of microbial resistance.


title: [Application of noninvasive positive pressure ventilation for sleep disordered breathing in China].

text: After more than 30 years of development, noninvasive positive pressure ventilation (NPPV) has become one of the main treatments of sleep-disordered breathing (SDB) in the world. It is the wide application of this technology that makes breakthrough in SDB treatment, and has given rise to a new interdisciplinary area sleep medicine. In China, sleep medicine started in late of 1980's, in recent years, with the development of economy and the improvement of recognization, the application of NPPV in Chinese market has become one of the fastest growing in the world. NPPV technology also extends gradually to the treatments of patients with respiratory failure due to different causes, such as severe acute respiratory syndrome and COPD.


title: The role of viruses in the aetiology of community-acquired pneumonia in adults.

text: BACKGROUND The role of viruses in community-acquired pneumonia may have been previously underestimated. We aimed to study the incidence and clinical characteristics of community-acquired pneumonia (CAP) due to respiratory viruses in adults adding PCR to routine conventional laboratory tests. METHODS Consecutive adult patients diagnosed of CAP from January 2003 to March 2004 were included. Conventional tests including cultures of blood, sputum, urine antigen detection of Streptococcus pneumoniae and Legionella pneumophila, and paired serologies were routinely performed. Nasopharyngeal swabs were processed for study of respiratory viruses through antigen detection by indirect immunofluorescence assay, isolation of viruses in cell culture and detection of nucleic acids by two independent multiplex RT-PCR assays. According to the aetiology, patients were categorized in 4 groups: group 1, only virus detected; group 2, only bacteria detected; group 3, viral and bacterial; and group 4, unkown aetiology. RESULTS Of 340 patients diagnosed with CAP, 198 had nasopharyngeal swabs available and were included in this study. Aetiology was established in 112 (57%) patients: group 1, n=26 (13%); group 2, n=66 (33%); group 3, n=20 (10%). The most common aetiological agent was S. neumoniae (58 patients, 29%), followed by respiratory viruses (46 patients, 23%). Forty-eight respiratory viruses were identified: influenza virus A (n=16), respiratory syncytial virus A (n=5), adenovirus (n=8), parainfluenza viruses (n=5), enteroviruses (n=1), rhinoviruses (n=8) and coronavirus (n=5). There were two patients coinfected by two respiratory viruses. Serology detected 6 viruses, immunofluorescence 8, viral culture 12, and PCR 45. For the viruses that could be diagnosed with conventional methods, the sensitivity and specificity of RT-PCR was 85% and 92%, respectively. The only clinical characteristic that significantly distinguished viral from bacterial aetiology was a lower number of leukocytes (P=0.004). CONCLUSION PCR revealed that viruses represent a common aetiology of CAP. There is an urgent need to reconsider routine laboratory tests for an adequate diagnosis of respiratory viruses, as clinical characteristics are unable to reliably distinguish viral from bacterial aetiology.


title: N-terminal domain of the murine coronavirus receptor CEACAM1 is responsible for fusogenic activation and conformational changes of the spike protein.

text: The mouse hepatitis virus (MHV) receptor (MHVR), CEACAM1, has two different functions for MHV entry into cells: binding to MHV spike protein (S protein) and activation of the S protein to execute virus-cell membrane fusion, the latter of which is accompanied by conformational changes of the S protein. The MHVR comprising the N-terminal and fourth domains [R1(1,4)] displays these two activities, and the N domain is thought to be critical for binding to MHV. In this study, we have addressed whether or not the N domain alone is sufficient for these activities. We examined three types of soluble form MHVR (soMHVR), one consisting of the N domain alone [soR1(1)], one with the N and second domains [soR1(1,2)], and one [soR1(1,4)] expressed by recombinant baculoviruses. We assessed the abilities of these three types of soMHVR to bind to MHV, activate fusogenicity, and induce conformational changes of the S protein. All three types of soMHVR similarly bound to MHV, as examined by a solid-phase binding assay and neutralized MHV infectivity. They also activated S protein fusogenicity and induced its conformational changes with similar levels of efficiency. However, R1(1) expressed on the BHK cell surface failed to serve as a receptor in spite of a sufficient level of expression. The inability of expressed R1(1) to work as a receptor was due to the inaccessibility of virions to R1(1); however, these were accessible using the MHVR-specific monoclonal antibody CC1. These results collectively indicated that the N domain retains all biological activities necessary for receptor function.


title: Requirement of the 5'-end genomic sequence as an upstream cis-acting element for coronavirus subgenomic mRNA transcription.

text: We have developed a defective interfering (DI) RNA containing a chloramphenicol acetyltransferase reporter gene, placed behind an intergenic sequence, for studying subgenomic mRNA transcription of mouse hepatitis virus (MHV), a prototype coronavirus. Using this system, we have identified the sequence requirement for MHV subgenomic mRNA transcription. We show that this sequence requirement differs from that for RNA replication. In addition to the previously identified requirement for an intergenic (promoter) sequence, additional sequences from the 5' end of genomic RNA are required for subgenomic mRNA transcription. These upstream sequences include the leader RNA and a spacer sequence between the leader and intergenic sequence, which is derived from the 5' untranslated region and part of gene 1. The spacer sequence requirement is specific, since only the sequence derived from the 5' end of RNA genome, but not from other MHV genomic regions or heterologous sequences, could initiate subgenomic transcription from the intergenic sequence. These results strongly suggest that the wild-type viral subgenomic mRNAs (mRNA2 to mRNA7) and probably their counterpart subgenomic negative-sense RNAs cannot be utilized for mRNA amplification. Furthermore, we have demonstrated that a partial leader sequence present at the 5' end of genome, which lacks the leader-mRNA fusion sequence, could still support subgenomic mRNA transcription. In this case, the leader sequences of the subgenomic transcripts were derived exclusively from the wild-type helper virus, indicating that the MHV leader RNA initiates in trans subgenomic mRNA transcription. Thus, the leader sequence can enhance subgenomic transcription even when it cannot serve as a primer for mRNA synthesis. These results taken together suggest that the 5'-end leader sequence of MHV not only provides a trans-acting primer for mRNA initiation but also serves as a cis-acting element required for the transcription of subgenomic mRNAs. The identification of an upstream cis-acting element for MHV subgenomic mRNA synthesis defines a novel sequence requirement for regulating mRNA synthesis in RNA viruses.


Feline immunodeficiency virus pathogenesis and development of a dual-subtype feline-immunodeficiency-virus vaccine.


title: Clinical Characterization of 162 COVID-19 patients in Israel: Preliminary Report from a Large Tertiary Center.

text: BACKGROUND In February 2020, the World Health Organisation designated the name COVID-19 for a clinical condition caused by a virus identified as a cause for a cluster of pneumonia cases in Wuhan, China. The virus subsequently spread worldwide, causing havoc to medical systems and paralyzing global economies. The first COVID-19 patient in Israel was diagnosed on 27 February 2020. OBJECTIVES To present our findings and experiences as the first and largest center for COVID-19 patients in Israel. METHODS The current analysis included all COVID-19 patients treated in Sheba Medical Center from February 2020 to April 2020. Clinical, laboratory, and epidemiological data gathered during their hospitalization are presented. RESULTS Our 162 patient cohort included mostly adult (mean age of 52 ± 20 years) males (65%). Patients classified as severe COVID-19 were significantly older and had higher prevalence of arterial hypertension and diabetes. They also had significantly higher white blood cell counts, absolute neutrophil counts, and lactate dehydrogenase. Low folic acid blood levels were more common amongst severe patients (18.2 vs. 12.9 vs. 9.8, P = 0.014). The rate of immune compromised patients (12%) in our cohort was also higher than in the general population. The rate of deterioration from moderate to severe disease was high: 9% necessitated non-invasive oxygenation and 15% were intubated and mechanically ventilated. The mortality rate was 3.1. CONCLUSIONS COVID-19 patients present a challenge for healthcare professionals and the whole medical system. We hope our findings will assist other providers and institutions in their care for these patients.


title: Making G8 leaders deliver: an analysis of compliance and health commitments, 1996-2006.

text: International health policy-makers now have a variety of institutional instruments with which to pursue their global and national health goals. These instruments range from the established formal multilateral organizations of the United Nations to the newer restricted-membership institutions of the Group of Eight (G8). To decide where best to deploy scarce resources, we must systematically examine the G8's contributions to global health governance. This assessment explores the contributions made by multilateral institutions such as the World Health Organization, and whether Member States comply with their commitments. We assessed whether G8 health governance assists its member governments in managing domestic politics and policy, in defining dominant normative directions, in developing and complying with collective commitments and in developing new G8-centred institutions. We found that the G8's performance improved substantially during the past decade. The G8 Member States function equally well, and each is able to combat diseases. Compliance varied among G8 Member States with respect to their health commitments, and there is scope for improvement. G8 leaders should better define their health commitments and set a one-year deadline for their delivery. In addition, Member States must seek WHO's support and set up an institution for G8 health ministers.


title: Global infectious disease surveillance and health intelligence.

text: Current concerns about the spread of infectious diseases, especially unexpected ("emerging") infections such as pandemic influenza or severe acute respiratory syndrome (SARS), have renewed focus on the critical importance of global early warning and rapid response. Although considerable progress has been made, many gaps remain. A number of the gaps can be addressed through increased political will, resources for reporting, improved coordination and sharing of information, raising clinicians' awareness, and additional research to develop more rigorous triggers for action. The increasing availability of communications and information technologies worldwide offers new opportunities for reporting even in low-capacity settings.


title: Is intralesional cidofovir worthwhile in juvenile recurrent respiratory papillomatosis?

text: OBJECTIVE To investigate the efficacy of intralesional cidofovir in the treatment of recurrent respiratory papillomatosis (RRP) in children. METHODS Prospective observational study of four consecutive children with RRP treated at an academic tertiary children's hospital. Laryngo-bronchoscopy was performed at three- to five-weekly intervals. Photodocumentation was obtained and disease severity assessed using an anatomical RRP severity score. Surgical debulking of large papillomas was then performed, and cidofovir (5 mg/ml) injected into any remaining papillomas as well as submucosally at the sites of resected papillomas. The efficacy of cidofovir was assessed by the change in papilloma severity score over the course of the treatment. RESULTS Complete disease remission was obtained in one patient, with a partial response seen in two others. One patient showed no significant response. The greatest beneficial effect was seen after the fourth cidofovir injection; however, two patients demonstrated a deterioration in severity scores after treatment was withheld at this point. Both responded well to further cidofovir injections. However, a clear plateau in the response to cidofovir was seen in all patients by the eighth injection. CONCLUSION Intralesional cidofovir may help control papilloma regrowth and reduce disease severity in many children with RRP. In most cases, cidofovir would appear to be less efficacious in causing disease eradication. There appears to be little evidence to support prolonged treatment regimes (i.e. more than eight treatments).


title: Chlorhexidine--pharmaco-biological activity and application.

text: OBJECTIVES Chlorhexidine (CHX) is one of the most widely used antiseptic, especially in dentistry. At low concentrations CHX is bacteriostatic and at high concentrations acts bactericidal causing cell death by cytolysis. In this study, we performed a systematic review of pharmaco-biological activity and application of CHX. MATERIALS AND METHODS Articles for inclusion in this review were retrieved from online databases PubMed/Medline. The selected papers were included in the present manuscript according to their relevance for the topic. RESULTS Totally 75 papers were enrolled in this research. CHX has strong biocidal activity against Gram-positive bacteria and weaker activity against Gram-negative bacteria. It is also active against yeasts, some dermatophytes and some lipophilic viruses. The most widely application CHX has found in dentistry and antisepsis. Numerous studies have confirmed the beneficial effects of CHX in reducing of plaque accumulation, in tooth caries, gingivitis, periodontitis and in alveolar osteitis. Unfortunately, CHX exhibits cytotoxic activity on human cells, can cause colorization of teeth and fillings, and its activity depends on the pH of the environment and the presence of organic substances. CONCLUSIONS CHX play a valuable role in the dentistry and antisepsis. However, it can also cause side effects, limiting its application time.


title: Mechanism of action of glycyrrhizic acid in inhibition of Epstein-Barr virus replication in vitro.

text: We report here that glycyrrhizic acid (GL), a component of licorice root (Glycyrrhiza radix), is active against EBV replication in superinfected Raji cells in a dose-dependent fashion. The IC(50) values for viral inhibition and cell growth were 0.04 and 4.8mM, respectively. The selectivity index (ratio of IC(50) for cell growth to IC(50) for viral DNA synthesis) was 120. Time of addition experiments suggested that GL interferes with an early step of EBV replication cycle (possibly penetration). GL had no effect on viral adsorption, nor did it inactivate EBV particles. Thus, GL represents a new class of anti-EBV compounds with a mode of action different from that of the nucleoside analogs that inhibit viral DNA polymerase.


title: Detection and differentiation of porcine epidemic diarrhoea virus and transmissible gastroenteritis virus in clinical samples by multiplex RT-PCR.

text: A multiplex reverse-transcriptase-PCR (RT-PCR) procedure was developed for the simultaneous detection of porcine epidemic diarrhoea virus (PEDV) and transmissible gastroenteritis virus (TGEV) in preweaning pigs with diarrhoea. The membrane gene of PEDV and the nucleocapsid gene of TGEV were chosen as targets. The PCR products of PEDV and TGEV had molecular sizes of 412 and 612 base pairs, respectively. Primers from PEDV did not react with any TGEV tested and vice versa. In addition, the primers did not react with other pig viruses. The multiplex RT-PCR was able to detect 10 tissue culture-infective doses 50 per cent (TCID50)/ml of PEDV or TGEV with each of the primer sets for PEDV and TGEV, respectively. The RNAS of PEDV and TGEV were detected directly in intestinal and faecal samples from pigs infected experimentally with either virus. The results of the assay correlated well with the results of virus isolation. None of the five control specimens was positive. PEDV was detected in 10 intestinal and nine faecal samples, and among the nine positive faecal samples two were culture-negative. TGEV was also detected in 10 intestinal and nine faecal samples, and among the nine positive faecal samples, three were culture-negative.


title: Prior infection and passive transfer of neutralizing antibody prevent replication of severe acute respiratory syndrome coronavirus in the respiratory tract of mice.

text: Following intranasal administration, the severe acute respiratory syndrome (SARS) coronavirus replicated to high titers in the respiratory tracts of BALB/c mice. Peak replication was seen in the absence of disease on day 1 or 2, depending on the dose administered, and the virus was cleared within a week. Viral antigen and nucleic acid were detected in bronchiolar epithelial cells during peak viral replication. Mice developed a neutralizing antibody response and were protected from reinfection 28 days following primary infection. Passive transfer of immune serum to naïve mice prevented virus replication in the lower respiratory tract following intranasal challenge. Thus, antibodies, acting alone, can prevent replication of the SARS coronavirus in the lung, a promising observation for the development of vaccines, immunotherapy, and immunoprophylaxis regimens.


title: Circulation of infectious bronchitis virus strains from Italy 02 and QX genotypes in Slovenia between 2007 and 2009.

text: Fourteen infectious bronchitis viruses (IBVs) detected in broiler, broiler breeder, and layer flocks in Slovenia between 2007 and 2009 were molecularly analyzed by reverse transcription-PCR and direct sequencing of the S1 gene. Phylogenetic analyses based on partial S1 gene sequences revealed that seven strains were classified as the Italy 02 genotype, a genetic group of IBV that has not previously been detected in Slovenia. Another seven strains were classified as the QX genotype. The results of phylogenetic analyses and epidemiologic investigations revealed that the genetic classification correlates with the geographic origins of the IBV strains. Greater genetic variability (maximum nucleotide and amino acid divergences were 4.8% and 9.9%, respectively) was observed within the Slovenian strains from the Italy 02 genotype detected between 2007 and 2009 than within strains from the QX genotype isolated in 2008 and 2009 (1.2% and 1.3%, respectively). The Slovenian strains classified as the Italy 02 genotype form a separate genetic cluster. These strains shared five specific amino acid substitutions that became fixed during the period of surveillance. Strains from the QX genotype that shared one specific amino acid substitution also form a separate genetic cluster.


title: [Early experience in laparoscopic radical prostatectomy using the laparoscopic device for umbilical access SILS Port].

text: Since 2007, various urological procedures have been performed with laparoendoscopic single-site surgery (LESS surgery), including nephrectomy, pyeloplasty, simple prostatectomy and, with the refinement of laparoscopic instrumentation, radical prostatectomy. This paper reports our initial experience in radical prostatectomy using the SILS Port from Covidiem and two lateral 5-mm trocars for triangulation. The SILS Port allows for accurate, simple insertion through a Hadson incision. The flexible port accommodates three 5-mm cannulas or two 5-mm cannulas and a 12-mm port for easier instrument exchange through a single incision. This approach decreases morbidity from bleeding, hernia and/or internal organ damage and improves cosmetic. One-port single-incision laparoscopy is part of the natural development of minimally invasive surgery. Future research is required to assess the intraoperative and postoperative benefits of LESS surgery as compared to standard laparoscopy.


title: Prevalence of abnormal lipid levels among youths --- United States, 1999-2006.

text: Cardiovascular disease (CVD) is the leading cause of death among adults in the United States. CVD risk factors, including abnormal lipid levels and elevated body mass index (BMI), often emerge during childhood and adolescence. In 2008, the American Academy of Pediatrics (AAP) established recommendations for targeted screening of youths aged >or=2 years for abnormal blood lipid levels. To provide prevalence data on abnormal lipid levels among youths, eligibility for lipid screening based on BMI, and eligibility for therapeutic lifestyle counseling among overweight youths, CDC analyzed results from the National Health and Nutrition Examination Survey (NHANES) for 1999-2006. This report describes the results of that analysis, which found that the prevalence of abnormal lipid levels among youths aged 12-19 years was 20.3%. This prevalence varied by BMI; 14.2% of normal weight youths, 22.3% of overweight and 42.9% of obese had at least one abnormal lipid level. Among all youths, 32% had a high BMI and therefore would be candidates for lipid screening under AAP recommendations. Given the high prevalence of abnormal lipid levels among youths who are overweight and obese in this study, clinicians should be aware of lipid screening guidelines, especially recommendations for screening youths who are overweight or obese.


title: Differential susceptibility of cultured neural cells to the human coronavirus OC43.

text: By using cell-type-specific markers and neural cultures derived from various areas of the nervous system, it has been possible to identify various interactions between OC43 virus and mouse oligodendrocytes, neurons, astrocytes, and fibroblasts. Neurons derived from dorsal root ganglia produced viral antigen and infectious virus. Astrocytes and fibroblasts both produced viral antigen but not infectious virus. Oligodendrocytes produced neither infectious virus nor viral antigen. Human embryo brain cells, including astrocytes, were susceptible to OC43 infection but did not produce infectious virus.


Newly identified respiratory viruses.


title: Evaluation of neck masses in adults.

text: Neck masses are often seen in clinical practice, and the family physician should be able to determine the etiology of a mass using organized, efficient diagnostic methods. The first goal is to determine if the mass is malignant or benign; malignancies are more common in adult smokers older than 40 years. Etiologies can be grouped according to whether the onset/duration is acute (e.g., infectious), subacute (e.g., squamous cell carcinoma), or chronic (e.g., thyroid), and further narrowed by patient demographics. If the history and physical examination do not find an obvious cause, imaging and surgical tools are helpful. Contrast-enhanced computed tomography is the initial diagnostic test of choice in adults. Computed tomography angiography is recommended over magnetic resonance angiography for the evaluation of pulsatile neck masses. If imaging rules out involvement of underlying vital structures, a fine-needle aspiration biopsy can be performed, providing diagnostic information via cytology, Gram stain, and bacterial and acid-fast bacilli cultures. The sensitivity and specificity of fine-needle aspiration biopsy in detecting a malignancy range from 77% to 97% and 93% to 100%, respectively.


Implications of infectious agents on results of animal experiments. Report of the Working Group on Hygiene of the Gesellschaft für Versuchstierkunde--Society for Laboratory Animal Science (GV-SOLAS).


title: High level transient expression of the murine coronavirus haemagglutinin-esterase.

text: We have expressed the murine coronavirus haemagglutinin-esterase protein in a vaccinia virus/T7 RNA polymerase system. The levels of expression observed are significantly higher than those found in virus-infected cells. The expressed protein has both receptor-destroying (esterase) and receptor-binding (haemadsorption) activities. The use of this system will greatly facilitate analysis of the structure-function relationships of this protein.


Trends in collection, use and disclosure of personal information in contemporary health research: challenges for research governance.


title: Biologic, antigenic, and full-length genomic characterization of a bovine-like coronavirus isolated from a giraffe.

text: Coronaviruses (CoVs) possess large RNA genomes and exist as quasispecies, which increases the possibility of adaptive mutations and interspecies transmission. Recently, CoVs were recognized as important pathogens in captive wild ruminants. This is the first report of the isolation and detailed genetic, biologic, and antigenic characterization of a bovine-like CoV from a giraffe (Giraffa camelopardalis) in a wild-animal park in the United States. CoV particles were detected by immune electron microscopy in fecal samples from three giraffes with mild-to-severe diarrhea. From one of the three giraffe samples, a CoV (GiCoV-OH3) was isolated and successfully adapted to serial passage in human rectal tumor 18 cell cultures. Hemagglutination assays, receptor-destroying enzyme activity, hemagglutination inhibition, and fluorescence focus neutralization tests revealed close biological and antigenic relationships between the GiCoV-OH3 isolate and selected respiratory and enteric bovine CoV (BCoV) strains. When orally inoculated into a BCoV-seronegative gnotobiotic calf, GiCoV-OH3 caused severe diarrhea and virus shedding within 2 to 3 days. Sequence comparisons and phylogenetic analyses were performed to assess its genetic relatedness to other CoVs. Molecular characterization confirmed that the new isolate belongs to group 2a of the mammalian CoVs and revealed closer genetic relatedness between GiCoV-OH3 and the enteric BCoVs BCoV-ENT and BCoV-DB2, whereas BCoV-Mebus was more distantly related. Detailed sequence analysis of the GiCoV-OH3 spike gene demonstrated the presence of a deletion in the variable region of the S1 subunit (from amino acid 543 to amino acid 547), which is a region associated with pathogenicity and tissue tropism for other CoVs. The point mutations identified in the structural proteins (by comparing GiCoV-OH3, BCoV-ENT, BCoV-DB2, and BCoV-Mebus) were most conserved among GiCoV-OH3, BCoV-ENT, and BCoV-DB2, whereas most of the point mutations in the nonstructural proteins were unique to GiCoV-OH3. Our results confirm the existence of a bovine-like CoV transmissible to cattle from wild ruminants, namely, giraffes, but with certain genetic properties different from those of BCoVs.


Are community-based interventions effective in identifying and responding to emerging zoonotic infectious diseases?


title: Infection of HeLa cells by avian infectious bronchitis virus is dependent on cell status.

text: To investigate the adaptation of avian infectious bronchitis virus (IBV) in a human cell line may be beneficial to understanding the potential mechanisms of coronavirus interspecies infection. The current study addressed the poor replication of IBV in the HeLa human cell line demonstrated in previous reports. We showed that IBV strains M41, H52, H120 and Gray could be propagated in HeLa cells with distinct cytopathic effect. The virus titre in freshly dispersed HeLa cells was 1000-fold higher than in cell monolayers. Trypsin was not the determinant for the viral replication, suggesting that the restriction of IBV replication in HeLa cells is the result of intracellular events rather than the binding to or fusion with host cells. These IBV strains replicated to an average titre of 10(3.4+/-0.2)/0.1 ml median tissue culture infectious doses in freshly dispersed HeLa cells and maintained this titre for the first 12 passages. Then an approximately 10-fold increase (10(4.20+/-0.19)/0.1 ml) occurred in passage 13, which was maintained to passage 16, after which there was another, bigger rise to 10(6.6+/-0.3)/0.1 ml in passage 17. This titre was maintained until passage 24 when the experiment was terminated. The IBV M41 S1 gene was amplified and sequenced for passages 0, 5 and 21. There was only one amino acid replacement in the S1 protein, in passage 21. The presence of sialic acid on HeLa cells contributed to efficient virus replication, while human aminopeptidase N was not involved in the infection. Haemagglutinin activity gradually reduced with increased passages. These results indicated that the virus adaptation would probably be determined by host cell modification such as receptor glycosylation and different receptor utilization instead of viral gene mutation.


title: Concurrent experimentally induced infection with Eimeria bovis and coronavirus in unweaned dairy calves.

text: Over a period of 3 summers, 21 colostrum-fed Holstein bull calves, 1 to 3 days old, were assigned to 7 replicates, each consisting of 3 calves. Within each replicate of 3 calves, 2 were selected at random, to be given 100,000 to 146,000 sporulated coccidia oocysts (principally Eimeria bovis) orally 60 hours after arrival at the college research farm. On the thirteenth day after coccidia inoculation, 1 of the 2 calves that had been given coccidia and the third calf that had not been inoculated, were given coronavirus by intranasal and oral routes. Calves were observed daily, and consistency of feces was scored visually. Nasal swab specimens for indirect immunofluorescent antibody testing for coronavirus and fecal samples for oocyst determination were obtained approximately every third day. Of 7 calves that were given only coronavirus, 3 developed diarrhea of short duration. Of 7 calves that were given only coccidia oocysts, 6 developed diarrhea. All 7 calves inoculated initially with coccidia and subsequently with coronavirus developed diarrhea. For 5 of 7 replicates, calves that were given coccidia and coronavirus developed diarrhea first. When overall severity, measured by fecal score and by blood in the feces, was compared, calves inoculated with coccidia followed by coronavirus were more severely affected (P less than 0.05) than were calves that were given only coronavirus. Calves that were given only coccidia oocysts appeared more severely affected than calves that were given only coronavirus, but differences were not significant.(ABSTRACT TRUNCATED AT 250 WORDS)


A Consequential Eight Years for Health Care and Public Health.


title: Intracellular location and translocation of silent and active poliovirus replication complexes.

text: Replication of poliovirus (PV) genomic RNA in HeLa cells has previously been found to start at distinct sites at the nuclear periphery. In the present study, the earliest steps in the virus replication cycle, i.e. the appearance and intracellular translocation of viral protein and negative-strand RNA prior to positive-strand RNA synthesis, were followed. During translation, positive-strand RNA and newly synthesized viral protein presented as a dispersed endoplasmic reticulum (ER)-like pattern. Concomitant with translation, individual PV vesicle clusters emerged at the ER and formed nascent replication complexes, which contained newly synthesized negative-strand RNA. The complexes rapidly moved centripetally, in a microtubule-dependent way, to the perinuclear area to engage in positive-strand viral RNA synthesis. Replication complexes made transcriptionally silent with guanidine/HCl followed the anterograde membrane pathway to the Golgi complex within the microtubule-organizing centre (MTOC), whereas replication complexes active in positive-strand RNA synthesis were retained at the nuclear periphery. If the silent replication complexes that had accumulated at the MTOC were released from the guanidine block, transcription was not readily resumed. Rather, positive-strand RNA was redistributed back to the ER to start, after a lag phase, translation, followed by negative- and positive-strand RNA synthesis in replication complexes migrating to the nuclear periphery. As some of the findings appear to be in contrast to events reported in cell-free guanidine-synchronized translation/transcription systems, implications for the comparison of in vitro systems with the living cell are discussed.


title: [First detection of human metapneumovirus in a child with respiratory syndrome in Hungary].

text: INTRODUCTION Human metapneumovirus was identified in 2001 as a respiratory-tract pathogen that has been classified as a new genera in family Paramyxoviridae. AIMS Molecular detection of human metapneumovirus in Hungary. MATERIALS AND METHODS Human metapneumovirus was identified in nasopharyngeal aspirate amplification of the viral fusion and nucleocapsid genes by reverse transcription-polymerase chain reaction followed by sequencing and phylogenetic analysis. RESULTS A 4 years-old girl with chronic respiratory syndrome chronically treated with anti-asthma drugs was admitted to hospital in November 2005 with acute respiratory syndrome and atelectasis. Nasopharyngeal aspirate was negative for common bacteria by culture and for influenza and coronavirus by reverse transcription-polymerase chain reaction. By contrast, specimen was positive by reverse transcription-polymerase chain reaction and was confirmed by sequencing both genes (nucleocapsid and fusion) of human metapneumovirus. Human metapneumovirus (HUN 05-L20) clustered into the subgroup B1 has the closest nucleotide similarity (98%) to JPS03-194 (AY530094) detected in Japan. CONCLUSIONS Human metapneumovirus contributes as an etiological agent of acute lower and upper respiratory tract infection especially in winter season in children with bronchiolitis, pneumonia or episodes of asthma exacerbation in Hungary, too.


Non-pharmaceutical interventions.


title: Rotavirus and coronavirus outbreak: etiology of annual diarrhea in Costa Rican children.

text: In Costa Rica, an annual outbreak of infant diarrheal disease (December and January) was reported since 1976, and rotavirus was incriminated later as the main etiological agent (1976-1981). Apparently the disease has not been systematically studied in Costa Rica after 1981. For that reason the occurrence of the outbreak was retrospectively documented for 1993-1995 and etiology was studied in 48 children treated for diarrhea at the Nacional Children Hospital (capital city of San Jose) during December, 1994 and January, 1995. Rotavirus (33%) and coronavirus (27%) were the main agents. To our knowledge, this is the first time that these viruses are incriminated in an outbreak of diarrhea.


NICE recommends photodynamic therapy for certain patients with macular degeneration.


title: Summary of Notifiable Diseases - United States, 2011.

text: The Summary of Notifiable Diseases - United States, 2011 contains the official statistics, in tabular and graphic form, for the reported occurrence of nationally notifiable infectious diseases in the United States for 2011. Unless otherwise noted, the data are final totals for 2011 reported as of June 30, 2012. These statistics are collected and compiled from reports sent by state health departments and territories to the National Notifiable Diseases Surveillance System (NNDSS), which is operated by CDC in collaboration with the Council of State and Territorial Epidemiologists (CSTE).


title: Immunofluorescence assay for detection of the nucleocapsid antigen of the severe acute respiratory syndrome (SARS)-associated coronavirus in cells derived from throat wash samples of patients with SARS.

text: An antigen detection assay for severe acute respiratory syndrome (SARS) coronavirus was established in this study by an indirect immunofluorescence test, which utilized cells derived from throat wash samples of patients with SARS and a rabbit serum that recognized the nucleocapsid protein of SARS-associated coronavirus (SARS-CoV) but not that of other human coronavirus tested. It detected SARS-CoV in 11 of 17 (65%) samples from SARS patients as early as day 2 of illness but in none of the 10 samples from healthy controls. Compared with other diagnostic modalities for detecting SARS-CoV, this assay is simpler, more convenient, and economical. It could be an alternative for early and rapid diagnosis, should SARS return in the future.


title: Evaluation of human monoclonal antibody 80R for immunoprophylaxis of severe acute respiratory syndrome by an animal study, epitope mapping, and analysis of spike variants.

text: In this report, the antiviral activity of 80R immunoglobulin G1 (IgG1), a human monoclonal antibody against severe acute respiratory syndrome coronavirus (SARS-CoV) spike (S) protein that acts as a viral entry inhibitor in vitro, was investigated in vivo in a mouse model. When 80R IgG1 was given prophylactically to mice at doses therapeutically achievable in humans, viral replication was reduced by more than 4 orders of magnitude to below assay limits. The essential core region of S protein required for 80R binding was identified as a conformationally sensitive fragment (residues 324 to 503) that overlaps the receptor ACE2-binding domain. Amino acids critical for 80R binding were identified. In addition, the effects of various 80R-binding domain amino acid substitutions which occur in SARS-like-CoV from civet cats, and which evolved during the 2002/2003 outbreak and in a 2003/2004 Guangdong index patient, were analyzed. The results demonstrated that the vast majority of SARS-CoVs are sensitive to 80R. We propose that by establishing the susceptibility and resistance profiles of newly emerging SARS-CoVs through early S1 genotyping of the core 180-amino-acid neutralizing epitope of 80R, an effective immunoprophylaxis strategy with 80R should be possible in an outbreak setting. Our study also cautions that for any prophylaxis strategy based on neutralizing antibody responses, whether by passive or active immunization, a genotyping monitor will be necessary for effective use.


Considerations on Colorectal Cancer Care in a COVID-19 Pandemic Epicenter.


title: A region of the coronavirus infectious bronchitis virus 1a polyprotein encoding the 3C-like protease domain is subject to rapid turnover when expressed in rabbit reticulocyte lysate.

text: In order to investigate the mechanisms involved in the processing of infectious bronchitis virus polyproteins, several candidate regions of the genome have been cloned and expressed in vitro. During these studies it was observed that the translation product encoded by one of these clones (pKT205) was poorly expressed. Biochemical and genetic analyses revealed that the basis for the poor expression was a post-translational event involving ubiquitination of the protein and degradation by an ATP-dependent system operating in the reticulocyte lysate used for the in vitro expression. Two independently acting regions which conferred instability were identified, one of which mapped to the predicted 3C protease domain, contained within the 5' end of the clone, while the other, more C-terminal region, was effective in conferring instability upon a heterologous protein to which it had been transferred. These regions may influence the stability of the authentic viral protein(s) in vivo and hence allow for the control of their expression and/or function at the level of proteolysis by cellular protease(s).


title: Structural studies on the Ebola virus matrix protein VP40 indicate that matrix proteins of enveloped RNA viruses are analogues but not homologues.

text: Matrix proteins are the driving force of assembly of enveloped viruses. Their main function is to interact with and polymerize at cellular membranes and link other viral components to the matrix-membrane complex resulting in individual particle shapes and ensuring the integrity of the viral particle. Although matrix proteins of different virus families show functional analogy, they share no sequence or structural homology, Their diversity is also evident in that they use a variety of late domain motifs to commit the cellular vacuolar protein sorting machinery to virus budding. Here, we discuss the structural and functional aspects of teh filovirus matrix protein VP40 and compare them to other known matrix protein structures from vesicular stomatitis virus adn retroviral matrix protein.


title: Concomitant cryptosporidia, coronavirus and parvovirus infection in a raccoon (Procyon lotor).

text: A juvenile raccoon (Procyon lotor) was found moribund near Fort Collins, Colorado (USA). Upon examination, the raccoon was dehydrated, had a mucopurulent oculonasal discharge and diarrhea, and was euthanized. Postmortem examination revealed emaciation, severe fibrinous gastroenteritis and a small, firm liver. Histopathological findings included blunting of villi, infiltration of lamina propria with neutrophils and plasma cells, and mild bronchopneumonia. Cryptosporidium sp. was demonstrated on intestinal villi and coronavirus and parvovirus were identified in feces. Fluorescent antibody test for rabies was negative and no evidence of canine distemper was found.


title: Crossing the nuclear envelope: hierarchical regulation of nucleocytoplasmic transport.

text: Transport of macromolecules between the nucleus and cytoplasm is a critical cellular process for eukaryotes, and the machinery that mediates nucleocytoplasmic exchange is subject to multiple levels of control. Regulation is achieved by modulating the expression or function of single cargoes, transport receptors, or the transport channel. Each of these mechanisms has increasingly broad impacts on transport patterns and capacity, and this hierarchy of control directly affects gene expression, signal transduction, development, and disease.


title: Phylogenetic and evolutionary relationships among torovirus field variants: evidence for multiple intertypic recombination events.

text: Toroviruses (family Coronaviridae, order Nidovirales) are enveloped, positive-stranded RNA viruses that have been implicated in enteric disease in cattle and possibly in humans. Despite their potential veterinary and clinical relevance, little is known about torovirus epidemiology and molecular genetics. Here, we present the first study into the diversity among toroviruses currently present in European swine and cattle herds. Comparative sequence analysis was performed focusing on the genes for the structural proteins S, M, HE, and N, with fecal specimens serving as sources of viral RNA. Sequence data published for animal and human torovirus variants were included. Four genotypes, displaying 30 to 40% divergence, were readily distinguished, exemplified by bovine torovirus (BToV) Breda, porcine torovirus (PToV) Markelo, equine torovirus Berne, and the putative human torovirus. The ungulate toroviruses apparently display host species preference. In phylogenetic analyses, all PToV variants clustered, while the recent European BToVs mostly resembled the New World BToV variant Breda, identified 19 years ago. However, we found ample evidence for recurring intertypic recombination. All newly characterized BToV variants seem to have arisen from a genetic exchange, during which the 3' end of the HE gene, the N gene, and the 3' nontranslated region of a Breda virus-like parent had been swapped for those of PToV. Moreover, some PToV and BToV variants carried chimeric HE genes, which apparently resulted from recombination events involving hitherto unknown toroviruses. From these observations, the existence of two additional torovirus genotypes can be inferred. Toroviruses may be even more promiscuous than their closest relatives, the coronaviruses and arteriviruses.


title: Persevering through a difficult time during the SARS outbreak in Toronto.

text: The purpose of this study was to describe the experience of persevering through a difficult time for patients, family members of patients, nurses, and allied health professionals during the severe acute respiratory syndrome outbreak. van Kaam's phenomenological research method, with the human becoming theory as the theoretical perspective, was used to gather and analyse data from 63 participants who agreed to describe a situation that illuminated their experience of persevering through a difficult time (either online or using a voicemail system). Data gathering occurred in early April 2003 in the midst of the severe acute respiratory syndrome outbreak in Toronto, Canada. The finding was the structural definition, persevering through a difficult time is dispiriting trepidation arising with witnessing suffering. It is a smothering connectedness with sequestering protection as unsettling contentment emerges amid unburdening hope. It sheds light on what is important for preparing for possible future outbreaks of this and other infectious diseases.


title: Self-reported frequent mental distress among adults--United States, 1993-2001.

text: Poor mental health is a major source of distress, disability, and social burden; in any given year, as many as one in five adults in the United States has a mental disorder. To identify differences among populations and factors contributing to poor mental health, CDC examined the prevalence of frequent mental distress (FMD) among U.S. adults by race/ethnicity, socioeconomic status (SES), and sex, by using aggregate data from Behavioral Risk Factor Surveillance System (BRFSS) surveys for 1993-2001. This report describes the results of that analysis, which indicated that the prevalence of FMD varied among racial/ethnic populations and increased substantially among whites and blacks. In addition, FMD was reported more frequently by women and by persons with low SES within each racial/ethnic population. Targeting adverse socioeconomic risk factors and improving access to mental health services might decrease FMD among adults and reduce racial/ethnic disparities in mental health.


title: A unique case of reversible myocardial ischemia in a hyperthyroid neonate.

text: A case of an 8-day-old preterm neonate with heart failure, pulmonary hypertension, and myocardial ischemia due to hyperthyroidism is reported. Treatment of the disease initially with b-blockers and, upon establishment of hyperthyroidism, with propylthiouracil reversed all cardiac abnormalities. Contrary to the rule, diagnosis of hyperthyroidism in the mother was established following the diagnosis of the condition in her baby. On long-term follow-up (10 years), the child has developed normally, remains euthyroid with normal electrocardiogram. To our knowledge, diagnosis and reversal of the above-mentioned ischemic abnormalities have not been previously reported in neonates.


title: Towards elimination of tuberculosis in New Zealand.

text: New Zealand could be the first country in the world to eliminate tuberculosis (TB). We propose a TB elimination strategy based on the eight-point World Health Organization (WHO) action framework for low incidence countries. Priority actions recommended by the WHO include 1) ensure political commitment, funding and stewardship for planning and essential services; 2) address the most vulnerable and hard-to-reach groups; 3) address special needs of migrants and cross-border issues; 4) identify active TB and undertake screening for latent tuberculosis infection (LTBI) in recent TB contacts and selected high-risk groups, and provide appropriate treatment; 5) optimise the prevention and care of drug-resistant TB; 6) ensure continued surveillance, programme monitoring and evaluation and case-based data management; 7) invest in research and new tools; and 8) support global TB prevention, care and control. In New Zealand, central government needs to take greater responsibility for TB policy and programme governance. Urgent action is required to prevent TB in higher risk groups including Māori communities, and to enable immigration screening to detect and treat LTBI. Clinical services need to be supported to implement new guidelines for LTBI that enable better targeting of screening and shorter, safer treatment regimens. Access to WHO recommended treatment regimens needs to be guaranteed for drug-resistant TB. Better use of existing data could better define priority areas for action and assist in the evaluation of current control activities. Access to GeneXpert® MTB-RIF near the point of care and whole genome sequencing nationally would greatly improve clinical and public health management through early identification of drug resistance and outbreaks. New Zealand already has a world-class TB research community that could be better deployed to assist high-incidence countries through research and training.


title: Gastrojejunal strictures after Roux-en-Y gastric bypass with a 21-MM circular stapler.

text: INTRODUCTION Intraluminal staplers for gastrojejunostomy construction during Roux-en-Y gastric bypass (RYGBP) may be associated with postoperative strictures. We analyzed outcomes of a transabdominal circular-stapled RYGBP with evaluation of short- and long-term anastomotic complications. METHODS All laparoscopic RYGBPs performed between January 2004 and December 2005 at an academic institution were reviewed. The gastrojejunostomy was created by using the transabdominal passage of a 21-mm intraluminal circular stapler into an antecolic, antegastric Roux limb. This retrospective chart review analyzes patient demographics, anastomotic complications, and weight loss. RESULTS Between January 2004 and December 2005, 159 patients underwent transabdominal circular-stapled RYGBP. Fifteen patients developed a stenosis at the gastrojejunostomy, all requiring endoscopic balloon dilatation. One of these patients required laparoscopic revision of the gastrojejunostomy. Eleven strictures occurred after 30 days, whereas only 4 strictures occurred within 30 days of surgery. Two marginal ulcerations were seen within 1 year of surgery. CONCLUSION Our 9.4% stricture rate parallels what has been reported in the literature. The majority of strictures were amenable to one endoscopic treatment session. Transabdominal circular-stapled gastrojejunostomy is a reproducible construct for use in bariatric surgery.


title: Piezoelectric immunosensor for SARS-associated coronavirus in sputum.

text: A piezoelectric immunosensor was developed for the detection of SARS-associated coronavirus (SARS-CoV) in sputum in the gas phase. Horse polyclonal antibody against SARS-CoV was bound onto the PZ crystal surface in an ordered orientation through protein A. The antigen sample was atomized into aerosol by an ultrasonator, by which the antibody on the crystal could specifically adsorb SARS antigen and the changed mass of crystal would lead a frequency shift. A frequency counter was employed to record the admittance frequency, and the plot of changed frequency was displayed on the computer. Under the optimized conditions, the frequency shifts were linearly dependent on antigen concentration in the range of 0.6-4 microg/mL. The device has good reproducibility (could be reused 100 times without detectable loss of activity), stability (the immunosensor was stable for more than two months when stored over silica gel blue at 4-6 degrees C), short analyzing time (less than 2 min), and specificity.


title: Evasion of antibody neutralization in emerging severe acute respiratory syndrome coronaviruses.

text: Molecular characterization of the severe acute respiratory syndrome coronavirus has revealed genetic diversity among isolates. The spike (S) glycoprotein, the major target for vaccine and immune therapy, shows up to 17 substitutions in its 1,255-aa sequence; however, the biologic significance of these changes is unknown. Here, the functional effects of S mutations have been determined by analyzing their affinity for a viral receptor, human angiotensin-converting enzyme 2 (hACE-2), and their sensitivity to Ab neutralization with viral pseudotypes. Although minor differences among eight strains transmitted during human outbreaks in early 2003 were found, substantial functional changes were detected in S derived from a case in late 2003 from Guangdong province [S(GD03T0013)] and from two palm civets, S(SZ3) and S(SZ16). S(GD03T0013) depended less on the hACE-2 receptor and was markedly resistant to Ab inhibition. Unexpectedly, Abs that neutralized most human S glycoproteins enhanced entry mediated by the civet virus S glycoproteins. The mechanism of enhancement involved the interaction of Abs with conformational epitopes in the hACE-2-binding domain. Finally, improved immunogens and mAbs that minimize this complication have been defined. These data show that the entry of severe acute respiratory syndrome coronaviruses can be enhanced by Abs, and they underscore the need to address the evolving diversity of this newly emerged virus for vaccines and immune therapies.


title: Recanalization of popliteal and infrapopliteal chronic total occlusions using Viance and CrossBoss crossing catheters: a multicenter experience from the XLPAD Registry.

text: BACKGROUND Chronic total occlusions (CTOs) are reported in up to 40% of patients with symptomatic peripheral arterial disease. The Viance Crossing catheter (Covidien) and the CrossBoss catheter (Boston Scientific) are novel devices that facilitate either true lumen or subintimal navigation across CTOs. The purpose of this study was to examine the acute procedural success of these devices for popliteal and below-the-knee (BTK) CTOs. METHODS Clinical and procedural outcome data between Sept 2010 and October 2013 were analyzed from the ongoing multicenter XLPAD registry. Technical success was defined as true lumen passage of the catheter, while procedural success was defined as successful vessel revascularization inclusive of subintimal passage and reentry. RESULTS Twenty-nine patients underwent 31 procedures, in which 37 lesions were treated with the Viance and CrossBoss catheters. Mean age of the group was 69.1 ± 10.7 years and 26 patients were male (90%). All patients (100%) had hypertension, 62% were diabetic, and 48% were active smokers. Critical limb ischemia was the indication for the procedure in 58% of cases; mean Rutherford class was 4.2 ± 1.2 for the entire cohort. Target CTOs included 14 anterior tibial, 9 posterior tibial, 5 peroneal, 1 tibio-peroneal trunk, and 8 popliteal artery lesions. Mean lesion length was 81 ± 64 mm; 15 lesions (41%) were severely calcified. True lumen passage was obtained in 24 lesions (65% technical success). Subintimal entry was achieved in 5 lesions (14%). Procedural success was achieved in 26 lesions (70%). Procedural failure was related to longer lesion length (P<.001), and mean length of failed lesions was 136 ± 65 mm. CONCLUSION Viance and CrossBoss catheters were associated with an overall 70% procedural success with predominantly true lumen crossing in a BTK-CTO cohort. CTO length remains an important determinant of device success.


Antiviral agents and corticosteroids in the treatment of severe acute respiratory syndrome (SARS).


The impact of covid-19 on business: how to deal with it.


title: Molecular cloning and nucleotide sequencing of the 3'-terminal genomic RNA of the porcine reproductive and respiratory syndrome virus.

text: The genomic RNA of a porcine reproductive and respiratory syndrome virus (PRRSV) isolate from the U.S.A., VR 2385 (ATCC), was copied into cDNA after priming with oligo(dT) and cloned into phage lambda. The cDNA clones representing the 3'-terminal genomic RNA of the virus were isolated and sequenced. The genome is a positive-stranded, polyadenylated RNA with an estimated size of 15 kb. Analysis of the resulting sequence identified three complete open reading frames (ORFs) with the potential to encode polypeptides with predicted M(r)s of 22.2K (ORF 5), 19.1K (ORF 6) and 13.6K (ORF 7). ORF 7, which is closest to the 3' end, is predicted to encode a highly basic nucleocapsid protein displaying 58% amino acid identity to the corresponding protein of the Lelystad virus (LV), a European PRRSV isolate. ORFs 6 and 5, preceding ORF 7, are each predicted to encode proteins containing several hydrophobic domains that are thought to be membrane-associated. The VR 2385 ORF 6 protein is the most conserved structural protein. It has 78% amino acid identity to the equivalent LV protein, and ORF 5 shares only 54% of its amino acid sequence. Northern blot analysis revealed a 3'-coterminal nested set of six subgenomic RNAs in VR 2385 virus-infected CRL 11171 cells. Our results indicate that VR 2385, like LV, is a member of the newly proposed arterivirus group. However, the striking genetic variation and the difference in pathogenicity between LV and VR 2385 suggest that the viruses causing PRRS in the U.S.A. and Europe are highly variable and they may represent different genotypes.


title: Xenotransplantation: an ethical dilemma.

text: PURPOSE OF REVIEW Xenotransplantation is an attractive alternative to allogeneic transplantation because of the intractable shortage of donor organs. The latest published developments in this area are discussed as well as recent publications regarding the continuing debates on the ethics of xenotransplantation and the safeguards that should be imposed to assure the minimization of risk to the patient and to human societies. RECENT FINDINGS Several laboratories are concentrating on the development of genetically manipulated donor pigs in order to bypass the immunological barriers against transplantation of porcine organs and cells into humans. Preliminary experiments with these new pig strains suggest that this strategy has a high likelihood of success with respect to hyperacute rejection. Ethical considerations have been extended to the establishment of regulatory, legal, and ethical frameworks that will be necessary to minimize the risk of xenotransplantation to the recipients, their families, and to the worldwide community. SUMMARY Xenotransplantation, because of the complexity of the medical, ethical and legal issues, will likely remain a controversial issue. If the scientific problems are solved, the decision to proceed with clinical application of this technique will depend on a collective decision guided by ethical, regulatory, and legal frameworks established by consensus.


[Acute bronchitis].


SARS: lessons to learn for GPs when handling a public health crisis.


title: Basic principles of real-time quantitative PCR.

text: Real-time quantitative PCR allows the sensitive, specific and reproducible quantitation of nucleic acids. Since its introduction, real-time quantitative PCR has revolutionized the field of molecular diagnostics and the technique is being used in a rapidly expanding number of applications. This exciting technology has enabled the shift of molecular diagnostics toward a high-throughput, automated technology with lower turnaround times. This article reviews the basic principles of real-time PCR and describes the various chemistries available: the double-stranded DNA-intercalating agent SYBR Green 1, hydrolysis probes, dual hybridization probes, molecular beacons and scorpion probes. Quantitation methods are discussed in addition to the competing instruments available on the market. Examples of applications of this important and versatile technique are provided throughout the review.


title: Exercise capacity and pulmonary function in hospital workers recovered from severe acute respiratory syndrome.

text: BACKGROUND Persistent impaired pulmonary function and functional capacity are common among survivors of severe acute respiratory syndrome (SARS). Whether the impairment was caused by SARS or pre-existing physical condition remains unclear. OBJECTIVE This study investigated the influence of SARS on exercise capacity and pulmonary function of previously healthy medical staff. METHODS Pulmonary function tests, including spirometry and carbon monoxide diffusing capacity (DLCO), as well as symptom-limited cardiopulmonary exercise testing (CPET) with an incremental protocol using an electronically braked cycle ergometer, were performed by 13 previously healthy hospital workers 14 months after SARS recovery. Other 14 age- and sex-matched healthy medical workers completed CPET simultaneously, and exercise capacities of these two groups were compared. RESULTS Most values of spirometry performed were within normal range. Only one showed mildly restrictive abnormality with decreased forced expiratory volume in 1 s (72.2% predicted) and forced vital capacity (68.1% predicted). Eight subjects had decreased DLCO levels (mean 79.37 +/- 7.73%), and low exercise capacity was noted in 9 subjects. Discordance in impairment of the measured DLCO and exercise capacity was revealed by comparison. Besides, there was no significant difference in results of CPET between subjects recovered from SARS and those never infected. CONCLUSIONS Minor pulmonary function defects as well as decreased exercise capacity were detected in previously healthy medical staff after recovering from SARS. No significant correlation between exercise capacity and pulmonary function was found.


title: Role of the thymus in streptococcal cell wall-induced arthritis and hepatic granuloma formation. Comparative studies of pathology and cell wall distribution in athymic and euthymic rats.

text: Systemic administration of an aqueous suspension of group A streptococcal cell wall fragments to susceptible rats induces acute and chronic polyarthritis, as well as noncaseating hepatic granulomas. To gain insight into the role of the thymus in the pathogenesis of this experimental model, pathologic responses and cell wall tissue distribution were compared in congenitally athymic rats (rnu/rnu) and their euthymic littermates (NIH/rnu). Within 24 h, both rat strains developed acute arthritis, characterized by polymorphonuclear leukocytic exudate in the synovium and joint spaces. This acute process was maximal at day 3 and gradually subsided. Beginning 2-3 wk after injection, the euthymic, but not the athymic, rats developed the typical exacerbation of arthritis, characterized by synovial cell hyperplasia with villus formation and T helper/inducer lymphocyte-rich mononuclear cell infiltration. This process eventually resulted in marginal erosions and destruction of periarticular bone and cartilage. Parallel development of acute and chronic hepatic lesions was observed. Bacterial cell wall antigen distribution and persistence were similar in the athymic and euthymic rats. Cell wall antigens were demonstrated in the cytoplasm of cells within subchondral bone marrow, synovium, liver, and spleen, coincident with the development of the acute lesions, and persisted in these sites, although in decreasing amounts, for the duration of the experiment. Our findings provide evidence that the acute and chronic phases of the experimental model are mechanistically distinct. The thymus and functional thymus derived-lymphocytes appear not to be required for the development of the acute exudative disease but are essential for the development of chronic proliferative and erosive disease. Induction of disease is dependent upon cell wall dissemination to and persistence in the affected tissues.