title: Confronting an influenza pandemic: ethical and scientific issues.

text: The prolonged concern over the potential for a global influenza pandemic to cause perhaps many millions of fatalities is a chilling one. After the SARS (severe acute respiratory syndrome) scares [1], attention has turned towards the possibility of an avian influenza virus hybridizing with a human influenza virus to create a highly virulent, as yet unknown, killer, on a scale unseen since the Spanish flu outbreak of 1918, which produced more fatalities than the Great War. In deciding how countries should react to this potential pandemic, individually and collectively, a reasonable and practical balance must be struck between the rights and obligations of individual citizens and protection of the wider community and, indeed, society as a whole. In this communication, ethical issues are discussed in the context of some of the scientific questions relating to a potential influenza pandemic. Among these issues are the rights and obligations of healthcare professionals, difficulties surrounding resource allocation, policies that have an impact on liberty and trade, when and how to introduce any vaccine or other form of mass treatment, global governance questions and the role of health policies in contemporary society. By considering these issues and questions in advance of an influenza, or indeed any other, pandemic commencing, countries can be better prepared to deal with the inevitably difficult decisions required during such events, rather than dusting down outdated previous plans, or making and implementing policy in an ad hoc manner with a resultant higher risk of adverse consequences.


title: Comparative evaluation of two severe acute respiratory syndrome (SARS) vaccine candidates in mice challenged with SARS coronavirus.

text: Two different severe acute respiratory syndrome (SARS) vaccine strategies were evaluated for their ability to protect against live SARS coronavirus (CoV) challenge in a murine model of infection. A whole killed (inactivated by beta-propiolactone) SARS-CoV vaccine and a combination of two adenovirus-based vectors, one expressing the nucleocapsid (N) and the other expressing the spike (S) protein (collectively designated Ad S/N), were evaluated for the induction of serum neutralizing antibodies and cellular immune responses and their ability to protect against pulmonary SARS-CoV replication. The whole killed virus (WKV) vaccine given subcutaneously to 129S6/SvEv mice was more effective than the Ad S/N vaccine administered either intranasally or intramuscularly in inhibiting SARS-CoV replication in the murine respiratory tract. This protective ability of the WKV vaccine correlated with the induction of high serum neutralizing-antibody titres, but not with cellular immune responses as measured by gamma interferon secretion by mouse splenocytes. Titres of serum neutralizing antibodies induced by the Ad S/N vaccine administered intranasally or intramuscularly were significantly lower than those induced by the WKV vaccine. However, Ad S/N administered intranasally, but not intramuscularly, significantly limited SARS-CoV replication in the lungs. Among the vaccine groups, SARS-CoV-specific IgA was found only in the sera of mice immunized intranasally with Ad S/N, suggesting that mucosal immunity may play a role in protection for the intranasal Ad S/N delivery system. Finally, the sera of vaccinated mice contained antibodies to S, further suggesting a role for this protein in conferring protective immunity against SARS-CoV infection.


title: Cell surface phenotype and human T lymphotropic virus type 1 antigen expression in 12 T cell lines derived from peripheral blood and cerebrospinal fluid of West Indian, Guyanese and African patients with tropical spastic paraparesis.

text: Twelve long-term cell lines were established from peripheral blood mononuclear cells (PBMC) or cerebrospinal fluid cells of patients with human T lymphotropic virus type I (HTLV-1) seropositive tropical spastic paraparesis (TSP) originating from the French West Indies, French Guyana or the Central African Republic. Most of these long-term interleukin-2-dependent cell lines exhibited a pattern characteristic of CD4(+)-activated T cells with high expression of CD2, CD3 and CD4 antigens, associated with a strong density of TAC and DR molecules. Nevertheless, in five cases CD8 expression was present at a significant level. HTLV-I antigens were never detected in uncultured PBMC, but they were expressed in a few cells after short-term culture and after 4 months the majority of the cells were HTLV-I positive, as demonstrated by indirect immunofluorescence (IF) using polyclonal or monoclonal anti-p19 and anti-p24 antibodies. Low and variable levels of reverse transcriptase activity were detected in supernatant fluids of these cell lines only after 4 months of culture, when at least 50% of the cells exhibited HTLV-I antigens by IF. However, numerous type C HTLV-I-like viral particles were detected, mostly in the extracellular spaces, with rare budding particles. Similar findings were found in three T cell lines derived from West Indian and African patients with adult T-cell leukaemia/lymphoma (ATLL). Differences in high Mr polypeptides were detected by Western blot in cell lysates when comparing TSP- or ATLL-derived T cell lines. Thus a signal of 62K was easily detectable in all the TSP lines, but not in the ATLL lines. In all cell lines bands corresponding to p53, p24 and p19 viral core polypeptides were present, as was the env gene-coded protein p46.


title: Gene profiles during root canal treatment in experimental rat periapical lesions.

text: The purpose of this study was to profile gene expression in periapical lesions during root canal treatment (RCT). Periapical lesions were induced experimentally by exposing the pulp in Sprague-Dawley rats. After 3 wk, the animals received root canal filling (RCF) and were sacrificed 1 or 4 wk later. From the periapical tissues, total RNA was extracted and processed for cDNA-microarray analysis. The lesions were histologically and radiographically confirmed to expand 4 wk after pulp exposure (inflammation phase) and to stabilize 4 wk after RCF (healing phase). In approximately 30,000 genes on the microarray, 203 genes were up-regulated to more than 5-fold (e.g., IL-1beta), and 864 genes were down-regulated to less than 20% of baseline level (e.g., caspase 8) in inflammation phase. Compared with inflammation phase, we found that 133 genes were up-regulated (e.g., IL-1alpha) and 50 genes were down-regulated (e.g., defensin alpha5) in healing phase. Corresponding to the gene expression profiles, accumulation of IL-1alpha and IL-1beta was observed in the periapical lesions by immunohistochemistry. These gene profiles might be useful in diagnosing the healing process of periapical lesions.


title: An outbreak of meningococcal disease in a secondary school--implications for public health practice.

text: This report describes briefly the management of three cases of meningococcal disease which all occurred within one week at a secondary school on the Central Coast of New South Wales in late winter 2003. The Central Coast health area has a population of approximately 300,000. Between 10 and 15 cases of meningococcal disease are notified to the Central Coast Public Health Unit each year. The three cases all presented to Gosford Hospital, Cases 1 and 2, both in Year 9, on Thursday 14 August 2003 and Case 3 in Year 8 on Friday 15 August 2003.


title: Dissection of Extracranial Internal Carotid Artery Due to Balloon Guiding Catheter Resulting in Asymptomatic Internal Carotid Artery Occlusion.

text: Dissection of the internal carotid artery (ICA) is a rare condition that accounts for a significant proportion of ischemic strokes in young adults. Iatrogenic dissection as a complication of neurointerventional procedures is a traumatic dissection which has been reported relatively rare in the literature. In this report, a case of dissection of the ICA is reported that was caused by repetitive movement of the balloon guiding catheter during stent-assisted thrombectomy (SAT), resulting in occlusion of the ICA.


title: Current developments in hernia repair; meshes, adhesives, and tacking.

text: Open and laparoscopic hernia surgery continues to evolve with new products allowing surgeons multiple choices in treating their patients. The evolution towards tension-free techniques in dealing with hernias requires that today's surgeons know the options available in meshes as well as fixation methods in order to have the best outcomes. In recent years, there has been a rapid expansion in the number of meshes available. Currently, there are numerous uncoated, coated, and biologic meshes in production that can be used in hernia repair. This paper will focus on the latest developments in coated meshes that allow for intra-abdominal placement as well as the different types of biologic meshes and their typical uses. Tacking devices for laparoscopic hernia repair now come in titanium as well as absorbable devices. AbsorbaTack™ (Covidien, Norwalk, CT) and Sorbafix™ (Davol, Warwick, RI) are two of the newest absorbable tacking devices thought to possibly benefit patients with decreased pain and long-term complications as compared with their titanium counterparts. Adhesives continue to be used more and more for hernia repair, especially in inguinal and paraesophageal hernia repairs. Tissucol™/Tisseel™ (Baxter, Deerfield, IL) and Evicel™ (Ethicon, Somerville, NJ) are two types of fibrin glues that are available for use in hernia repair. Practitioners using these biologic adhesives think there is less pain compared with tacking.


title: High-sensitivity detection methods for low-abundance RNA species: applications for functional genomics research.

text: Gene expression analysis has facilitated a more complete understanding of the molecular biology of cellular processes and how variations of RNA expression are useful for the classification of various diseases. Furthermore, recent analysis of a variety of noncoding RNAs, such as microRNAs, has demonstrated that these RNAs play an important role in many cellular events, including cell differentiation and death, and may also serve as biological markers for disease. Besides helping in the understanding of diseases, RNA analysis is used in drug discovery, patient prognosis and treatment evaluation. One obstacle left to overcome is the amount of material required for the analysis, particularly when trying to extract information from precious, limited, clinical samples. Here we review the many approaches scientists take to either amplify the amount of RNA or amplify the signal generated from small amounts of RNA.


title: [SARS (Severe Acute Respiratory Syndrome). Emergent transmissible disease].

text: Of the reemergent transmissible diseases of the past decades, SARS is probably not the last to express the alterations occurring in the relationships of the human being with its global ecosystem. The life of contemporary man is characterized, among others, by a huge thirst for traveling, for varied reasons, consequence of the globalization process. SARS virus, mutant belonging to Coronaviridae, occurred in one of the most densely populated areas of the world. There are two main moments marking the reemergence and evolution of SARS: firstly, the onset of the epidemic in China in November 2002 followed by the worldwide spread of the epidemiological process, and secondly the discovery of SARS virus as a mutant of coronaviruses in March-April 2003 in USA, Canada, and Hong Kong. The possibilities of general and special prevention, and particularly vaccine prevention are likely to bring this disease under control.


title: Severe acute respiratory syndrome: clinical and laboratory manifestations.

text: Severe acute respiratory syndrome (SARS) is a recently emerged infectious disease with significant morbidity and mortality. An epidemic in 2003 affected 8,098 patients in 29 countries with 774 deaths. The aetiological agent is a new coronavirus spread by droplet transmission. Clinical and general laboratory manifestations included fever, chills, rigor, myalgia, malaise, diarrhoea, cough, dyspnoea, pneumonia, lymphopenia, neutrophilia, thrombocytopenia, and elevated serum lactate dehydrogenase (LD), alanine aminotransferase (ALT) and creatine kinase (CK) activities. Treatment has been empirical; initial potent antibiotic cover, followed by simultaneous ribavirin and corticosteroids, with or without pulse high-dose methylprednisolone, have been used. The postulated disease progression comprises (1) active viral infection, (2) hyperactive immune response, and (3) recovery or pulmonary destruction and death. We investigated serum LD isoenzymes and blood lymphocyte subsets of SARS patients, and found LD1 activity as the best biochemical prognostic indicator for death, while CD3+, CD4+, CD8+ and natural killer cell counts were promising predictors for intensive care unit (ICU) admission. Plasma cytokine and chemokine profiles showed markedly elevated Th1 cytokine interferon (IFN)-gamma, inflammatory cytokines interleukin (IL)-1beta, IL-6 and IL-12, neutrophil chemokine IL-8, monocyte chemoattractant protein-1 (MCP-1), and Th1 chemokine IFN-gamma-inducible protein-10 (IP-10) for at least two weeks after disease onset, but there was no significant elevation of inflammatory cytokine tumor necrosis factor (TNF)-alpha and anti-inflammatory cytokine IL-10. Corticosteroid reduced IL-8, MCP-1 and IP-10 concentrations from 5-8 days after treatment. Measurement of biochemical markers of bone metabolism demonstrated significant but transient increase in bone resorption from Day 28-44 after onset of fever, when pulse steroid was most frequently given. With tapering down of steroid therapy, there was a decrease in bone resorption marker together with an increase in bone formation markers round Day 50, suggesting that some of the bone loss might be reversed. Our research studies on the chemical pathology and clinical immunology of SARS should have implications for the pathophysiology and therapy of this potentially lethal infection.


title: Recommendations for a standardised educational program in robot assisted gynaecological surgery: Consensus from the Society of European Robotic Gynaecological Surgery (SERGS).

text: Background The Society of European Robotic Gynaecological Surgery (SERGS) aims at developing a European consensus on core components of a curriculum for training and assessment in robot assisted gynaecological surgery. Methods A Delphi process was initiated among a panel of 12 experts in robot assisted surgery invited through the SERGS. An online questionnaire survey was based on a literature search for standards in education in gynaecological robot assisted surgery. The survey was performed in three consecutive rounds to reach optimal consensus. The results of this survey were discussed by the panel and led to consensus recommendations on 39 issues, adhering to general principles of medical education. Results On review there appeared to be no accredited training programs in Europe, and few in the USA. Recommendations for requirements of training centres, educational tools and assessment of proficiency varied widely. Stepwise and structured training together with validated assessment based on competencies rather than on volume emerged as prerequisites for adequate and safe learning. An appropriate educational environment and tools for training were defined. Although certification should be competence based, the panel recommended additional volume based criteria for both accreditation of training centres and certification of individual surgeons. Conclusions Consensus was reached on minimum criteria for training in robot assisted gynaecological surgery. To transfer results into clinical practice, experts recommended a curriculum and guidelines that have now been endorsed by SERGS to be used to establish training programmes for robot assisted surgery.


title: What role for human rhinoviruses in the lower respiratory tract?

text: Human rhinoviruses (HRV) usually cause upper airway infections. However, viral replication in the tracheobronchial tree has been disclosed, although its clinical role is poorly known. We evaluated the prevalence of HRV in 159 bronchoalveolar lavages from 88 patients and describe a lung transplant recipient with a high HRV load in association with acute rejection. HRV was detected in 22/88 patients (25.0%): 7/18 lung transplant recipients, 11/41 immunocompetent, and 4/29 immunocompromised (p = n.s.). No lung disease was significantly associated with HRV positivity. It should be recommended to include HRV in the virological diagnostic work-up of lower respiratory specimens to elucidate their role.


title: Elicitation of immunity in mice after immunization with the S2 subunit of the severe acute respiratory syndrome coronavirus.

text: The S2 domain of the severe acute respiratory syndrome coronavirus (SARS-CoV) spike (S) protein is responsible for fusion between virus and target cell membranes, and is expected to be immungenic. In this study, we investigated the immune responses against the S2 subunit in BALB/c mice, which were vaccinated either with plasmid DNA encoding the S2 domain (residues 681-1120), the recombinant S2 fragment (residues 681-980) in incomplete Freund's adjuvant, or with inactivated SARS-CoV. The increased number of specific cytotoxic cells (CTLs) and the high titer of specific antibody showed stimulation of both arms of the immune system in these groups. The shift in cytokines suggested that Th1-polarized immune response was induced by plasmid pCoVS2, meanwhile the Th2-dominant response was induced by recombinant S2 fragment and inactivated vaccine. However, the titer of neutralizing antibodies was only detectable in mice immunized with inactivated virus, but not with pCoVS2 plasmid. Taken together, the S2 domain could induce specific cellular immune response and a high level of total IgG but little neutralizing antibodies against infection by SARSCoV.


title: Laparoscopic sleeve gastrectomy for morbid obesity with natural orifice specimen extraction (NOSE).

text: OBJECTIVE An experience with laparoscopic sleeve gastrectomy using the natural orifice specimen extraction (NOSE) technique. BACKGROUND Bariatric surgery is nowadays the only long term effective obesity treatment method. METHODS Twenty one consecutive patients underwent laparoscopic sleeve gastrectomy with the use of natural orifice specimen extraction (NOSE) in the Surgical Clinic of Faculty Hospital Ostrava between May 2012 and August 2012. Inclusion criteria were the body mass index (BMI) higher than 35 kg/m2 or higher than 32 kg/m2 accompanied with relevant comorbidities. RESULTS Among 21 patients in this series, there were three men (14.3%) and 18 women (85.7%). Their mean age was 40.9±10.2 years. Their mean preoperative BMI was 40.4±4.6 kg/m2. No patient had previous bariatric surgery, one patient had laparoscopic fundoplication. All operations were completed laparoscopically with no conversions to an open procedure. In two cases, laparoscopic cholecystectomy was performed and the gallbladder was extracted along with the gastric specimen by transgastric approach. CONCLUSION Laparoscopic sleeve gastrectomy is a safe and effective bariatric procedure with low morbidity and mortality. Based on our initial experiences it could be an indication for NOSE with transgastric approach. Obese patients would benefit from this approach due to the elimination of wound complications (Tab. 2, Fig. 3, Ref. 22).


title: The 2016 Infusion Therapy Standards of Practice.

text: Approximately every 5 years, the Infusion Nurses Society publishes evidence-based practice standards. This article provides an overview of the process used in standards development, describes the format of the standards, and provides a short summary of selected standards as applied to home care. The Standards are an important document that should be available to every home care organization that provides home infusion therapy.


title: Molecular evolution analysis and geographic investigation of severe acute respiratory syndrome coronavirus-like virus in palm civets at an animal market and on farms.

text: Massive numbers of palm civets were culled to remove sources for the reemergence of severe acute respiratory syndrome (SARS) in Guangdong Province, China, in January 2004, following SARS coronavirus detection in market animals. The virus was identified in all 91 palm civets and 15 raccoon dogs of animal market origin sampled prior to culling, but not in 1,107 palm civets later sampled at 25 farms, spread over 12 provinces, which were claimed to be the source of traded animals. Twenty-seven novel signature variation residues (SNVs) were identified on the spike gene and were analyzed for their phylogenetic relationships, based on 17 sequences obtained from animals in our study and from other published studies. Analysis indicated that the virus in palm civets at the live-animal market had evolved to infect humans. The evolutionary starting point was a prototype group consisting of three viral sequences of animal origin. Initially, seven SNV sites caused six amino acid changes, at positions 147, 228, 240, 479, 821, and 1080 of the spike protein, to generate low-pathogenicity viruses. One of these was linked to the first SARS patient in the 2003-2004 period. A further 14 SNVs caused 11 amino acid residue changes, at positions 360, 462, 472, 480, 487, 609, 613, 665, 743, 765, and 1163. The resulting high-pathogenicity groups were responsible for infections during the so-called early-phase epidemic of 2003. Finally, the remaining six SNVs caused four amino acid changes, at positions 227, 244, 344, and 778, which resulted in the group of viruses responsible for the global epidemic.


title: Expression of the recombinant anchorless N-terminal domain of mouse hepatitis virus (MHV) receptor makes hamster of human cells susceptible to MHV infection.

text: Mouse hepatitis virus (MHV) receptor, the receptor for the murine coronavirus MHV, was expressed in MHV-resistant hamster and human cells as a series of mutant, recombinant glycoproteins with carboxy-terminal deletions lacking the cytoplasmic tail, transmembrane domain, and various amounts of the immunoglobulin constant-region-like domains. The soluble receptor glycoproteins containing the N-terminal virus-binding domain were released into the supernatant medium and inactivated the infectivity of MHV-A59 virions in a concentration-dependent manner. Surprisingly, some of the anchorless glycoproteins were found on the plasma membranes of transfected cells by flow cytometry, and these cells were rendered susceptible to infection with three strains of MHV. Thus, in the cells in which the anchorless, recombinant receptor glycoprotein is synthesized, some of the protein is bound to an unidentified moiety on the plasma membrane, which allows it to serve as a functional virus receptor.


title: Human metapneumovirus: a newly emerging respiratory pathogen.

text: PURPOSE OF REVIEW The present review focuses on the recent discovery, and clinical and epidemiological features of a virus associated with respiratory tract infections. RECENT FINDINGS In June 2001, researchers in The Netherlands reported the discovery of a virus associated with respiratory tract disease in infants and children. Genetic studies of this newly discovered viral pathogen revealed that it was a paramyxovirus. This virus is the first human pathogen in the Genus Metapneumovirus and was called human metapneumovirus (hMPV). hMPV may be a common respiratory virus. Seroepidemiologic studies in The Netherlands suggested that, by age 5 years, nearly all individuals have been exposed to hMPV. The results of several studies suggest that hMPV may account for about 10% of respiratory tract infections in which a common respiratory virus, such as respiratory syncytial virus, or influenza or parainfluenza viruses, could not be detected. hMPV has been detected in patients with either upper or lower respiratory tract disease, or both. Symptoms associated with hMPV include cough, dyspnea, wheeze, and hypoxia. This newly recognized pathogen has been detected in children and adults. Epidemiological findings suggest that it may circulate worldwide and may have a seasonal distribution. SUMMARY hMPV is a newly emerging respiratory pathogen and may be the cause of a significant proportion of both upper and lower respiratory tract infection in infants, children, and adults.


title: [Detection of severe acute respiratory syndrome probable patients' virus RNA in Hangzhou by using a two loci and a modified nested real-time reverse transcription polymerase chain reaction].

text: OBJECTIVE To detect the RNA of severe acute respiratory syndrome virus (SARS-CoV) by using reverse transcription polymerase chain reaction (RT-PCR) targeted for a two loci and a modified nested real-time RT-PCR as to improving the reliability and sensitivity of tests. METHODS A nested RT-PCR was used for detecting one fragment of SARS-CoV RNA in oropharyngeal swabs from 3 SARS probable patients, 4 SARS suspect patients and other 27 patients with fever in Hangzhou, and the nested RT-PCR product from one SARS probable patient was sequenced. Meanwhile in these 3 SARS probable patients, other three RT-PCR methods, including a hemi-nested RT-PCR targeted for another fragment of SARS-CoV RNA, a real-time RT-PCR and a modified nested real-time RT-PCR, were employed to detect SARS-CoV RNA. RESULTS Two positives were found in the 3 SARS probable patients, and none positive in 4 SARS suspect patients and other 27 patients with fever, using the nested RT-PCR. The sequence of the nested RT-PCR product from one SARS probable patient was identified with the counterpart of SARS-CoV genomes published in public database. The results of the hemi-nested RT-PCR, the real-time RT-PCR and the modified nested real-time RT-PCR in the 3 SARS patients were consistent with the one of the nested RT-PCR. During detecting specimen with low copies of RNA, a weak positive signal was produced after about 35 cycles in the real-time RT-PCR, but a strong positive signal was found only after 10 cycles in the modified nested real-time RT-PCR. CONCLUSION It might improve the reliability of test by employing RT-PCR targeted for two or more fragments in SARS-CoV genome. The modified nested real-time RT-PCR might have higher sensitivity than the routine real-time RT-PCR.


Assessment of neonatal tetanus elimination in Malawi.


title: An Ubiquitin-like Motif in ASK1 Mediates its Association with and Inhibition of the Proteasome.

text: Linear polyubiquitin is processed at LRLRGG sequences by deubiquitinating enzymes to make free monomeric ubiquitin. This LRLRGG ubiquitin-like motif is found in a limited number of mammalian non-ubiquitin proteins, including the MAP3K Apoptosis Signal-Regulating Kinase-1 (ASK1), which activates MAPK signaling pathways. The c-terminus of ASK1 binds to the 19S cap of the proteasome allowing ASK1 to phosphorylate and inhibit proteasomal activity. We investigated whether the ubiquitin-like sequence in the c-terminus of ASK1 mediates its association with and inhibition of the proteasome. To test this we generated ASK1 with substitutions or deletions in this ubiquitin-like domain and examined the activation of cellular signaling and the association of ASK1 with the 19S cap of the proteasome. We show that ASK1 mutants have reduced association with the 19S cap of the proteasome, reduced capacity to inhibit the proteasome, and diminished ability to inhibit TNF-induced NF-κB activation. Mutant forms of ASK1 also had reduced capacity to activate JNK signaling, suggesting that the ubiquitin-like motif in ASK1 is also important for coordinating the balance between JNK and NF-κB signaling. Together these results demonstrate that the ubiquitin-like sequence of ASK1 is important for binding to and inhibition of the proteasome, and for the coordinated activation of cellular NF-κB and JNK signaling.


Article.


title: Serological survey of virus infection among wild house mice (Mus domesticus) in the UK.

text: The serological prevalence of 13 murine viruses was surveyed among 103 wild-caught and 51 captive-bred house mice (Mus domesticus), originating from several trapping locations in northwest England, using blood samples obtained during routine health screening of an established wild mouse colony. A high proportion of recently caught wild mice were seropositive for mouse hepatitis virus (86%), mouse cytomegalovirus (79%), mouse thymic virus (78%), mouse adenovirus (68%), mouse parvovirus (59%) and minute virus of mice (41%). Seroprevalences of lymphocytic choriomeningitis virus (LCMV), orthopoxvirus, reovirus-3 and murid herpesvirus 4 (MuHV-4, also called murine gamma-herpesvirus [MHV-68]) were low (3-13%), and no animals were seropositive to Sendai virus, pneumonia virus or polyomavirus. Seroprevalence in wild-caught animals that had been in captivity for over six months was generally consistent with the range found in recently caught wild animals, while seroprevalence was generally much lower in captive-bred mice despite no attempt to prevent viral spread. A notable exception to this was LCMV, which appeared to have spread efficiently through the captive population (both captive-bred and wild-caught animals). Given the known viral life cycles in laboratory mice, it appears that viral persistence in the host was an important contributing factor in the spread of infection in captivity.


title: Differential effects of human and porcine interleukin 2 on natural killing (NK) activity of newborn piglets and adult pigs lymphocytes.

text: The in vitro effects of human or porcine Interleukin 2 (IL2) on Natural Killing (NK) activity were studied with blood lymphocytes of newborn piglets and of adult pigs. Large volumes of porcine IL2 were prepared by PHA stimulation of irradiated mesenteric lymph node cells and, following purification by gel-filtration chromatography, the apparent molecular weight of IL2 was 15,000 Da. Purified human and porcine IL2 as well as recombinant human IL2 were found to increase markedly NK activity of lymphocytes derived from adult animals. However, although newborn piglets-derived lymphocytes are sensitive to crude porcine IL2 supernatants (and to Interferon alpha as previously shown) their low NK activity is unaffected by purified IL2. These data suggest therefore the existence of differences in the ontogenic development of the lymphokines responsiveness of porcine NK cells.


title: Trends and prediction in daily incidence of novel coronavirus infection in China, Hubei Province and Wuhan City: an application of Farr's law.

text: BACKGROUND The recent outbreak of novel coronavirus (2019-nCoV) has infected tens of thousands of patients in China. Studies have forecasted future trends of the incidence of 2019-nCoV infection, but appeared unsuccessful. Farr's law is a classic epidemiology theory/practice for predicting epidemics. Therefore, we used and validated a model based on Farr's law to predict the daily-incidence of 2019-nCoV infection in China and 2 regions of high-incidence. METHODS We extracted the 2019-nCoV incidence data of China, Hubei Province and Wuhan City from websites of the Chinese and Hubei health commissions. A model based on Farr's law was developed using the data available on Feb. 8, 2020, and used to predict daily-incidence of 2019-nCoV infection in China, Hubei Province and Wuhan City afterward. RESULTS We observed 50,995 (37,001 on or before Feb. 8) incident cases in China from January 16 to February 15, 2020. The daily-incidence has peaked in China, Hubei Providence and Wuhan City, but with different downward slopes. If no major changes occur, our model shows that the daily-incidence of 2019-nCoV will drop to single-digit by February 25 for China and Hubei Province, but by March 8 for Wuhan city. However, predicted 75% confidence intervals of daily-incidence in all 3 regions of interest had an upward trend. The predicted trends overall match the prospectively-collected data, confirming usefulness of these models. CONCLUSIONS This study shows the daily-incidence of 2019-nCoV in China, Hubei Province and Wuhan City has reached the peak and was decreasing. However, there is a possibility of upward trend.


title: Long-term haemodialysis survival.

text: Haemodialysis (HD) treatment for end-stage renal disease bears a poor prognosis. We present a case of a patient who, apart from two transplant periods lasting 8 months in all, was treated with conventional in-centre HD three times a week and who survived for 41 years. Patients should be aware that there is no theoretical upper limit for patient survival on HD.


title: Immunization of mice with a DNA vaccine based on severe acute respiratory syndrome coronavirus spike protein fragment 1.

text: According to data in GenBank, a gene encoding SARS spike protein fragment 1 (S1) was synthesized. After recombination with an immunostimulatory sequence (ISS), the gene was cloned into the plasmid pIRES to produce pIRES-ISS-S1. On confirmation of the expression of S1 protein by indirect immunofluorescence assay (IFA), after the transfection of pIRES-ISS-S1 into BHK-21 cells, the DNA vaccine was repeatedly administrated to BALB/c mice. CD4+ and CD8+ spleen T lymphocytes were analyzed by flow cytometry (FCM) to evaluate T cell-mediated immune responses, the antigen-specific responses of T cells were evaluated by cytotoxic T lymphocyte (CTL) assay, and the level of IgG in antisera from immunized mice was determined by enzyme-linked immunosorbent assay. Results showed that the counts of spleen CD4+ and CD8+ T lymphocytes were increased, that the T cell-mediated immune responses showed antigen specificity, and that IgG was significantly induced with DNA vaccines pIRES-ISS-S1 and pIRES-S1 at titers of 1:320 and 1:160, respectively. These results are promising for the protective immunization of humans.


title: Comparison of dexmedetomidine and propofol used for drug-induced sleep endoscopy in patients with obstructive sleep apnea syndrome.

text: UNLABELLED Backround: Surgical operations are alternative treatments in persons with Obstructive Sleep Apnea Syndrome who cannot tolerate continuous positive airway pressure therapy. Drug-Induced Sleep Endoscopy is a method with which somnolence is pharmacologically induced and collapse is evaluated through nasal endoscopy in patients with Obstructive Sleep Apnea Syndrome. AIMS We aimed to evaluate efficiency of dexmedetomidine or propofol used for sedation in patients undergoing drug-induced sleep endoscopy. METHODS A total of 40 patients aged between 18 and 65 years old in the ASA STATUS I-II group were included in the study. After premedicatıon wıth midazolam 0.05 mg/kg intravenously, patients were randomly divided into two groups and administered intravenous (iv) propofol with the loading dose of 0.7 mg/kg for 10 minutes, followed 0.5 mg/kg/h infusion (Group P); or dexmedetomidine with the loading dose of 1 mcg/kg for 10 minutes, followed by 0.3 mcg/kg/h infusion (Group D). Haemodynamic and respiratuary parameters, Bispectral index score, Ramsey sedation score, time to achieve sufficient sedation, surgeon's and patients' satisfaction, postoperative Aldrete score and side effects were recorded. RESULTS Time to achieve sufficient sedation, Bispectral index scores at 5, 10 and 15th. minutes intraoperatively, first Aldrete score in the recovery room, SpO2 values and respiratory rates all over the surgical procedure and in the recovery room were found lower in Group P (P<0.05). Bispectral index scores, mean arterial pressure and heart rate in the recovery room were significantly lower in Group D (P<0.05). CONCLUSION Dexmedetomidine may be preferred as a safer agent with respecting to respiratory function compared with propofol in obstructive sleep apnea patients who known to be susceptible to hypoxia and hypercarbia.


title: Identification of two epitopes in the carboxyterminal 15 amino acids of the E1 glycoprotein of mouse hepatitis virus A59 by using hybrid proteins.

text: cDNA fragments coding for the carboxy terminus of the E1 envelope glycoprotein from mouse hepatitis virus A59, a coronavirus, were cloned into the bacterial expression vector pEX. Clones expressing E1 antigenic determinants were selected with a polyclonal anti-E1 antibody and used for immunization of rabbits and for affinity purification of existing polyclonal antisera. Immunofluorescence testing and immunoperoxidase labeling of coronavirus-infected cells showed that these reagents were monospecific for E1. In addition, by using hybrid proteins containing different lengths of the E1 carboxy terminus to affinity-purify a polyclonal antiserum against E1, we have been able to define two epitopes within the last 15 amino acid residues of the protein. These epitope-specific antibodies bind to E1 in Golgi and perinuclear membranes as well as to budding viruses; they do not, however, label the plasma membrane or the membranes of post-Golgi vesicles transporting virions to the cell surface.


title: Persistent memory CD4+ and CD8+ T-cell responses in recovered severe acute respiratory syndrome (SARS) patients to SARS coronavirus M antigen.

text: The membrane (M) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) is a major glycoprotein with multiple biological functions. In this study, we found that memory T cells against M protein were persistent in recovered SARS patients by detecting gamma interferon (IFN-gamma) production using ELISA and ELISpot assays. Flow cytometric analysis showed that both CD4+ and CD8+ T cells were involved in cellular responses to SARS-CoV M antigen. Furthermore, memory CD8+ T cells displayed an effector memory cell phenotype expressing CD45RO- CCR7- CD62L-. In contrast, the majority of IFN-gamma+ CD4+ T cells were central memory cells with the expression of CD45RO+ CCR7+ CD62L-. The epitope screening from 30 synthetic overlapping peptides that cover the entire SARS-CoV M protein identified four human T-cell immunodominant peptides, p21-44, p65-91, p117-140 and p200-220. All four immunodominant peptides could elicit cellular immunity with a predominance of CD8+ T-cell response. This data may have important implication for developing SARS vaccines.


title: Bilayer dermal matrix for the treatment of painful burn scars.

text: UNLABELLED  Background. Outcomes following burn injury have improved considerably in recent years due to early excision and skin grafting. Despite a reduction in late burn complications, up to 70% of patients experience long-term pain at both the injured area and the skin-grafted scar. Current therapies do not ameliorate these symptoms for a majority of these patients. This report presents initial results of a new technique using a bilayer dermal substitute (Integra™, [Integra LifeSciences, Plainsboro, NJ]) for revision of painful scars. METHODS This is a prospective cohort of six patients treated at Grady Memorial Hospital Burn Center (Atlanta, GA) from 2008 to 2009. Burn patients undergoing multi-modality pain therapy for chronic, painful burns were identified and consented for intervention. All patients underwent operative excision of the painful scar and placement of a bilayer dermal matrix (Integra). After 3 weeks the silicone mesh was removed, excessive granulation tissue was debrided, and a new split-thickness skin graft was placed on the wound. Prescription history and a patient questionnaire were used to collect data. RESULTS All patients noticed improvement of symptoms post-operatively. Two patients (33%) experienced complete reduction of scar pain and three (50%) discontinued consumption of pain medications. At a mean follow up of 13 months, the average amount of narcotic-based pain medications decreased from 63.3 MED ([morphine equivalent per day], range: 30-135) before treatment to 7.5 MED (range: 0-20) postoperatively (P < 0.05). CONCLUSION Scar excision, interval placement of a bilayer dermal matrix, and subsequent skin grafting is a new technique that can improve, and in some cases ameliorate, burn scar related pain. .


title: Zika Virus Disease: Case Report and Review of Literature.

text: Zika virus (ZIKV) infection and its associated complications have become a significant public health concern. Zika virus is a Flavivirus, and is transmitted to humans by Aedes species mosquitoes. In May 2015, the World Health Organization reported the first locally acquired transmission of ZIKV in Brazil, the first case in the Western Hemisphere. There have also been reports of increased incidence of microcephaly and other neurologic complications associated with ZIKV infection, as well as a 20-fold increase in the incidence of Guillain-Barre Syndrome during ZIKV outbreaks. In light of the clusters of microcephaly and neurologic manifestations in areas of ZIKV transmission, the World Health Organization declared ZIKV and associated complications an international public health emergency. The clinical presentations of ZIKV disease resemble many common infections, making the diagnosis of ZIKV disease a clinical challenge. Patients with ZIKV infection may have other coinfections that can distract clinicians from suspecting ZIKV disease in their patients. Complications from ZIKV, although uncommon, may be severe. Therefore, it is important for health care providers to remain alert and up to date in the efforts to control this emerging global health crisis. It is critical for physicians and other providers to know the growing list of countries and territories on the US Centers for Disease Control and Prevention list of nations with endemic ZIKV disease and to work closely with local health departments to prevent spread. In this article, we present a case of ZIKV disease in a pediatric patient and review of the current literature.


title: BSGIE survey on COVID-19 and gastrointestinal endoscopy in Belgium : results and recommendations.

text: Background and aims With the first wave of the COVID-19 pandemic declining, activities in the gastrointestinal clinic are being recommenced after a period of stringent measures. Since a second COVID-19 wave is not entirely ruled out health care professionals might remain faced with the need to perform endoscopic procedures in patients with a confirmed positive or unknown COVID-19 status. With this report we aim to provide a practical relevant overview of preparation and protective measures for gastroenterologists based on the currently available guidelines and our local experience and results of a national Belgian survey, to guarantee a fast recall of an adequate infection prevention if COVID-19 reoccurs. Methods From the 23rd of March 2020 and the 13th of May 2020 we performed a Pubmed, Embase and Medline search, resulting in 37 papers on COVID-19 and endoscopy. Additionally, we combined these data with data acquired from the national BSGIE survey amongst Belgian gastroenterologists. Results Based on 72 completed surveys in both university and non-university hospitals, the results show (1) a dramatic (<20%) or substantial (<50%) decrease of normal daily endoscopy in 74% and 22% of the units respectively, (2) a difference in screening and protective measures between university and non-university hospitals. These findings were subsequently compared with the current guidelines. Conclusion Based on new data from the BSGIE survey and current guidelines we tried to realistically represent the current COVID-19 trends in protective measures, screening and indications for endoscopy and to provide a practical overview as preparation for a possible second wave.


title: Stable association of herpes simplex virus with target membranes is triggered by low pH in the presence of the gD receptor, HVEM.

text: Using a liposome-binding assay, we investigated the requirements for activation of herpes simplex virus (HSV) into a state capable of membrane interaction. Virions were mixed with liposomes along with the ectodomain of one of three gD receptors (HVEMt, nectin-1t, or nectin-2t) and incubated under different pH and temperature conditions. Virions failed to associate with liposomes in the presence of nectin-1 or nectin-2 at any temperature or pH tested. In contrast, HVEMt triggered association of HSV with liposomes at pH 5.3 or 5.0 when incubated at 37 degrees C, suggesting that HVEM binding and mildly acidic pH at a physiological temperature provide coactivation signals, allowing virus association with membranes. Virions incubated with HVEMt at 37 degrees C without liposomes rapidly lost infectivity upon exposure to pH 5.0, suggesting that these conditions lead to irreversible virus inactivation in the absence of target membranes. Consistent with the idea that soluble receptor molecules provide a trigger for HSV entry, HVEMt promoted virus entry into receptor-deficient CHO K1 cells. However, in B78H1 cells, HVEMt promoted virus entry with markedly lower efficiency. Interestingly, HSV entry into receptor-bearing CHO K1 cells has been shown to proceed via a pH-dependent manner, whereas HSV entry into receptor-bearing B78H1 cells is pH independent. Based on these observations, we propose that the changes triggered by HVEM and mildly acidic pH that allow liposome association are similar or identical to changes that occur during pH-dependent HSV entry.


docid: fsvueakk

title: Proinflammatory cytokines and viral respiratory disease in pigs.

text: Swine influenza virus (SIV), porcine respiratory coronavirus (PRCV) and porcine reproductive and respiratory syndrome virus (PRRSV) are enzootic viruses causing pulmonary infections in pigs. The first part of this review concentrates on known clinical and pathogenetic features of these infections. SIV is a primary respiratory pathogen; PRCV and PRRSV, on the contrary, tend to cause subclinical infections if uncomplicated but they appear to be important contributors to multifactorial respiratory diseases. The exact mechanisms whereby these viruses cause symptoms and pathology, however, remain unresolved. Classical studies of pathogenesis have revealed different lung cell tropisms and replication kinetics for each of these viruses and they suggest the involvement of different lung inflammatory responses or mediators. The proinflammatory cytokines interferon-alpha (IFN-alpha), tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) have been shown to play key roles in several respiratory disease conditions. The biological effects of these cytokines and their involvement in human viral respiratory disease are discussed in the second part of this review. The third part summarises studies that were recently undertaken in the authors' laboratory to investigate the relationship between respiratory disease in pigs and bioactive lung lavage levels of IFN-alpha, TNF-alpha and IL-1 during single and combined infections with the above viruses. In single SIV infections, typical signs of swine "flu" were tightly correlated with an excessive and coordinate production of the 3 cytokines examined. PRCV or PRRSV infections, in contrast, were subclinical and did not induce production of all 3 cytokines. Combined infections with these 2 subclinical respiratory viruses failed to potentiate disease or cytokine production. After combined inoculation with PRCV followed by bacterial lipopolysaccharide, both clinical respiratory disease and TNF-alpha/IL-1 production were markedly more severe than those associated with the respective single inoculations. Taken together, these data are the first to demonstrate that proinflammatory cytokines can be important mediators of viral respiratory diseases in pigs.


title: Lack of antibodies to coronaviruses in a captive cheetah (Acinonyx jubatus) population.

text: Cheetahs (Acinonyx jubatus) (n = 40) were tested by means of an immunofluorescent test (IFT) for the presence of antibodies to the feline coronavirus group. All cheetahs tested negatively and this was further confirmed by virus serum neutralisation.


title: Viral myocarditis and dilated cardiomyopathy: mechanisms, manifestations, and management.

text: Viral infection of the heart is relatively common and usually of little consequence. It can, however, lead to substantial cardiac damage and severe acute heart failure. It can also evolve into the progressive syndrome of chronic heart failure. Recent studies have gone some way towards unravelling the complex mechanisms underlying the heart muscle damage that occurs after viral infection. These studies have lent support to both immune and viral mediated (independent of an immune response) cardiac damage. Acute myocarditis can present in various ways, and it may be a cause of sudden death in an otherwise healthy young adult. New treatments for viral heart disease are awaited. In the meanwhile, the haemodynamic support of patients with acute left ventricular failure caused by myocarditis should be aggressive, to allow for the possibility of spontaneous recovery. Contemporary trials of treatment in chronic heart failure secondary to dilated cardiomyopathy support the use of angiotensin converting enzyme inhibitors, beta adrenoceptor blockers, and spironolactone in such patients.


A multistate outbreak of Salmonella enterica serotype Saintpaul infections linked to mango consumption: a recurrent theme.


Why China needs an NIH.


title: Comparative host gene transcription by microarray analysis early after infection of the Huh7 cell line by severe acute respiratory syndrome coronavirus and human coronavirus 229E.

text: The pathogenesis of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) at the cellular level is unclear. No human cell line was previously known to be susceptible to both SARS-CoV and other human coronaviruses. Huh7 cells were found to be susceptible to both SARS-CoV, associated with SARS, and human coronavirus 229E (HCoV-229E), usually associated with the common cold. Highly lytic and productive rates of infections within 48 h of inoculation were reproducible with both viruses. The early transcriptional profiles of host cell response to both types of infection at 2 and 4 h postinoculation were determined by using the Affymetrix HG-U133A microarray (about 22,000 genes). Much more perturbation of cellular gene transcription was observed after infection by SARS-CoV than after infection by HCoV-229E. Besides the upregulation of genes associated with apoptosis, which was exactly opposite to the previously reported effect of SARS-CoV in a colonic carcinoma cell line, genes related to inflammation, stress response, and procoagulation were also upregulated. These findings were confirmed by semiquantitative reverse transcription-PCR, reverse transcription-quantitative PCR for mRNA of genes, and immunoassays for some encoded proteins. These transcriptomal changes are compatible with the histological changes of pulmonary vasculitis and microvascular thrombosis in addition to the diffuse alveolar damage involving the pneumocytes.


docid: rgzxihbp

title: Rapid and sensitive method using multiplex real-time PCR for diagnosis of infections by influenza a and influenza B viruses, respiratory syncytial virus, and parainfluenza viruses 1, 2, 3, and 4.

text: Laboratory diagnosis of viral respiratory infections is generally performed by virus isolation in cell culture and immunofluorescent assays. Reverse transcriptase PCR is now recognized as a sensitive and specific alternative for detection of respiratory RNA viruses. A rapid real-time multiplex PCR assay was developed for the detection of influenza A and influenza B viruses, human respiratory syncytial virus (RSV), parainfluenza virus 1 (PIV1), PIV2, PIV3, and PIV4 in a two-tube multiplex reaction which used molecular beacons to discriminate the pathogens. A total of 358 respiratory samples taken over a 1-year period were analyzed by the multiplex assay. The incidence of respiratory viruses detected in these samples was 67 of 358 (19%) and 87 of 358 (24%) by culture and real-time PCR, respectively. Culture detected 3 influenza A virus, 2 influenza B virus, 57 RSV, 2 PIV1, and 2 PIV3 infections. All of these culture-positive samples and an additional 5 influenza A virus, 6 RSV, 2 PIV1, 1 PIV2, 1 PIV3, and 3 PIV4 infections were detected by the multiplex real-time PCR. The application of real-time PCR to clinical samples increases the sensitivity for respiratory viral diagnosis. In addition, results can be obtained within 6 h, which increases clinical relevance. Therefore, use of this real-time PCR assay would improve patient management and infection control.


Department of Health.


title: The coronavirus transmissible gastroenteritis virus causes infection after receptor-mediated endocytosis and acid-dependent fusion with an intracellular compartment.

text: Aminopeptidase N is a species-specific receptor for transmissible gastroenteritis virus (TGEV), which infects piglets, and for the 229E virus, which infects humans. It is not known whether these coronaviruses are endocytosed before fusion with a membrane of the target cell, causing a productive infection, or whether they fuse directly with the plasma membrane. We have studied the interaction between TGEV and a cell line (MDCK) stably expressing recombinant pig aminopeptidase N (pAPN). By electron microscopy and flow cytometry, TGEV was found to be associated with the plasma membrane after adsorption to the pAPN-MDCK cells. TGEV was also observed in endocytic pits and apical vesicles after 3 to 10 min of incubation at 38 degrees C. The number of pits and apical vesicles was increased by the TGEV incubation, indicating an increase in endocytosis. After 10 min of incubation, a distinct TGEV-pAPN-containing population of large intracellular vesicles, morphologically compatible with endosomes, was found. A higher density of pAPN receptors was observed in the pits beneath the virus particles than in the surrounding plasma membrane, indicating that TGEV recruits pAPN receptors before endocytosis. Ammonium chloride and bafilomycin A1 markedly inhibited the TGEV infection as judged from virus production and protein biosynthesis analyses but did so only when added early in the course of the infection, i.e., about 1 h after the start of endocytosis. Together our results point to an acid intracellular compartment as the site of fusion for TGEV.


The Cancer Problem.


title: RNA-cleaving properties of human apurinic/apyrimidinic endonuclease 1 (APE1).

text: We have recently identified apurinic/apyrimidinic endonuclease 1 (APE1) as an endoribonuclease that cleaves c-myc mRNA in vitro and regulates c-myc mRNA levels and half-life in cells. This study was undertaken to further unravel the RNA-cleaving properties of APE1. Here, we show that APE1 cleaves RNA in the absence of divalent metal ions and, at 2 mM, Zn(2+), Ni(2+), Cu(2+), or Co(2+) inhibited the endoribonuclease activity of APE1. APE1 is able to cleave CD44 mRNA, microRNAs (miR-21, miR-10b), and three RNA components of SARS-corona virus (orf1b, orf3, spike) suggesting that, when challenged, it can cleave any RNAs in vitro. APE1 does not cleave strong doublestranded regions of RNA and it has a strong preference for 3' of pyrimidine, especially towards UA, CA, and UG sites at single-stranded or weakly paired regions. It also cleaves RNA weakly at UC, CU, AC, and AU sites in single-stranded or weakly paired regions. Finally, we found that APE1 can reduce the ability of the Dicer enzyme to process premiRNAs in vitro. Overall, this study has revealed some previously unknown biochemical properties of APE1 which has implications for its role in vivo.


title: Motor vehicle-related death rates--United States, 1999-2005.

text: In 2005, the most recent year for which data are available, 45,520 deaths in the United States were related to motor vehicles. A Healthy People 2010 objective calls for reducing the rate of deaths related to motor vehicles to 9.2 per 100,000 population from a baseline of 15.6 in 1998. To assess progress toward the Healthy People objective and to examine characteristics of motor vehicle--related death rates, CDC analyzed data from the National Vital Statistics System (NVSS) for the period 1999--2005. This report summarizes the results of that analysis, which determined that, during 1999--2005, although annual age-adjusted motor vehicle--related death rates overall were nearly unchanged (range: 15.2--15.7 per 100,000 population), substantial differences were observed by state, U.S. Census region, sex, race, and age group. Among states, the average annual death rate ranged from 7.9 per 100,000 population in Massachusetts to 31.9 in Mississippi. Among regions, the rate ranged from 9.8 per 100,000 population in the Northeast to 19.5 in the South. The rate for men (21.7 per 100,000 population) was more than double the rate for women (9.4); the rate for American Indians/Alaska Natives (27.2) was nearly twice the rate for whites (15.7) and blacks (15.2), and the rate for persons aged 15--24 years (26.8) was 74% higher than the average annual rate overall (15.4). Additional analysis and research to determine the causes of geographic and demographic variations in motor vehicle--related deaths might result in more effective targeted interventions among the states, regions, and populations at greatest risk.


title: Tracing the origin and co-phylogeny of the caliciviruses.

text: Caliciviruses infect a wide range of mammalian hosts and include the genus Norovirus, the major cause of food-borne viral gastroenteritis in humans. Using publicly available sequence data and phylogenetic analysis tools, the origins and virus-host co-phylogeny of these viruses were investigated. Here, evidence is presented in support of host switching by caliciviruses, but showing that zoonotic transfer does not appear to have occurred in the history of these viruses. The age or demography of the caliciviruses cannot yet be estimated with any firm degree of support, but further studies of this family, as new dated sequences become available, could provide key information of importance to human health and in understanding the emergence of food-borne disease.


title: Molecular analysis of Brazilian infectious bronchitis field isolates by reverse transcription-polymerase chain reaction, restriction fragment length polymorphism, and partial sequencing of the N gene.

text: Molecular analysis of 15 Brazilian infectious bronchitis virus (IBV) isolates, obtained from clinical outbreaks of the disease in chickens (broilers or layers) in the state of Minas Gerais (Brazil) between 1972 and 1989, is reported. Using the N protein gene as target, IBVs were analyzed by reverse transcription-polymerase chain reaction/restriction fragment length polymorphism (RT-PCR/RFLP) with the restriction enzymes AvaII, HphI, Sau96I, and Tsp509I and cDNA sequencing. Results obtained from those isolates were compared to 19 sequences available in GenBank. N gene RFLP profiles, cDNA sequences, and predicted amino acid composition were used for the construction of dendrograms. Brazilian isolates were grouped into one distinct group. Identity of predicted N protein amino acid composition varied from 45% (between isolates G and 208) up to 99% (PM 1 and PM2), and, when compared to the other IBVs, the amino acid identity was from 42% (Q3/88 and G) up to 97% (D41 and PM1). The great genetic diversity was shown to occur before the official use of vaccination in Brazil and has remained thereafter.


title: Mouse hepatitis virus nucleocapsid protein as a translational effector of viral mRNAs.

text: The mouse hepatitis virus (MHV) nucleocapsid protein stimulated translation of a chimeric reporter mRNA containing an intact MHV 5'-untranslated region and the chloramphenicol acetyltransferase (CAT) coding region. The nucleocapsid protein binds specifically the tandemly repeated-UCYAA- of the MHV leader. This RNA sequence is the same as the intergenic motif found in the genome RNA. Preferential translation of viral mRNA in MHV infected cells is stimulated in part by this interaction and represents a specific, positive translational control mechanism employed by coronaviruses.


Lowered tuberculosis notifications and deterred health care seeking during the SARS epidemic in Hong Kong.


title: Sialic acid functions in enterovirus 70 binding and infection.

text: The interaction of viruses with host cell receptors is the initial step in viral infection and is an important determinant of virus host range, tissue tropism, and pathogenesis. The complement regulatory protein decay-accelerating factor (DAF/CD55) is an attachment receptor for enterovirus 70 (EV70), a member of the Picornaviridae, commonly associated with an eye infection in humans known as acute hemorrhagic conjunctivitis. In early work, the EV70 receptor on erythrocytes, responsible for its hemagglutinating activity, was shown to be sensitive to neuraminidase, implying an essential role for sialic acid in virus attachment. Here, we extend these results to show that cell surface sialic acid is required for EV70 binding to nucleated cells susceptible to virus infection and that sialic acid binding is important in productive infection. Through the use of site-directed mutagenesis to eliminate the single N-linked glycosylation site of DAF and of a chimeric receptor protein in which the O-glycosylated domain of DAF was replaced by a region of the HLA-B44 molecule, a role in EV70 binding for the sialic acid residues of DAF was excluded, suggesting the existence of at least one additional, sialylated EV70-binding factor at the cell surface. Treatment of cells with metabolic inhibitors of glycosylation excluded a role for the N-linked oligosaccharides of glycoproteins but suggested that O-linked glycosylation is important for EV70 binding.


title: SARS update: Winter, 2003 to 2004.

text: Transmission of severe acute respiratory syndrome (SARS) related coronavirus (CoV) appears to be heterogeneous. Most transmission occurs through large droplets, but there is some evidence of spread through aerosol transmission. Proper use of personal protective equipment (PPE) and scrupulous attention to hand hygiene are critical to reducing transmission of SARS, especially in health care facilities. Surgical masks have been shown to be highly effective in reducing transmission when N-95 respirators are unavailable. Health care workers bore a large burden of illness during the 2002 to 2003 SARS epidemic. Occupational health nurses working in health care agencies will play a key role in preparing for a re-emergence of the disease. Occupational health nurses should "bookmark" the Centers for Disease Control and Prevention website for Public Health Guidance for Community-Level Preparedness and Response to SARS: www.cdc.gov/ncidod/sars/clinicalguidance.htm for easy retrieval and reference.


title: Disease-resistant genetically modified animals.

text: Infectious disease adversely affects livestock production and animal welfare, and has impacts upon both human health and public perception of livestock production. The authors argue that the combination of new methodology that enables the efficient production of genetically-modified (GM) animals with exciting new tools to alter gene activity makes the applications of transgenic animals for the benefit of animal (and human health) increasingly likely. This is illustrated through descriptions of specific examples. This technology is likely to have specific application where genetic variation does not exist in a given population or species and where novel genetic improvements can be engineered. These engineered animals would provide valuable models with which to investigate disease progression and evaluate this approach to controlling the disease. The authors propose that the use of GM animals will complement the more traditional tactics to combat disease, and will provide novel intervention strategies that are not possible through the established approaches.


title: Identification of canine coronavirus strains from feces by S gene nested PCR and molecular characterization of a new Australian isolate.

text: A nested PCR (nPCR) assay for the detection of canine coronavirus (CCV) in fecal samples is described. The target sequence for the assay was a 514-bp fragment within the spike (S) glycoprotein gene. The sensitivity of the assay is extremely high, detecting as little as 25 50% tissue culture infective doses per g of unprocessed feces. A clinical trial using dogs challenged orally with CCV SA4 and CCV NVSL was used to compare viral isolation and the nPCR assay as detection techniques over a 2-week period of infection. Virus isolation detected CCV shedding from day 4 to 9 postchallenge, while the nPCR assay detected CCV shedding from day 4 to 13 postchallenge. Cloning and sequencing of the nPCR assay product enabled investigation of the evolutionary relationships between strains within the S gene. The simple and rapid procedure described here makes this assay an ideal alternative technique to electron microscopy and viral isolation in cell culture for detection of CCV shedding in feces. The described assay also provides a method of identifying new strains of CCV without the complicated and time-consuming practice of raising antibodies to individual strains. This is illustrated by the identification, for the first time, of an Australian isolate of CCV (UWSMN-1).


title: [Physiological and pathological consequences of a presence of germ line stem cells in adult tissues].

text: Various therapheutic strategies employing stem cells have been proposed as the alternative, effective methods for therapy of multitude diseases, difficult to treat using standard, well-established methods. Advancing regenerative medicine, which is becoming a novel branch of clinical medicine, has high hopes of stem cells which could be used in treatment of injuried organs such as myocardium after heart infarction, brain after stroke, spinal cord after mechanical injury as well as in treatment of diabetes and Parkinson disease. Application of embryonic stem cells, harvested from developing embryos, is highly controversial. Hence, the stem/primitive cells isolated from adult tissuses are considered to be an optimal source of cells for therapy. Recently our research team has isolated a population of very primitive stem cells from adult tissues (very small embryonic-like stem cells - VSELs) that show several embryonic-like features. These cells can become an alternative and more ethical source of the stem cells for therapy when compared to those isolated from the developing embryos.


title: Severe acute respiratory syndrome: a preliminary review.

text: Severe acute respiratory syndrome (SARS) is a new disease of unknown aetiology and probably a new strain of coronavirus is thought to be responsible for the disease. After originating from south-east China it has spread to several countries across the world. Patients of SARS suffer from fever with cough and dyspnoea. The virus spreads by droplet to nearby contacts and has high tendency to spread to the healthcare providers. Since the aetiology is not yet clear exact treatment is not yet defined and hence prevention is of utmost importance.


docid: omsuolly

title: [Post-traumatic stress disorder in convalescent patients of severe acute respiratory syndrome: (1)H-MRS study].

text: OBJECTIVE To ascertain the metabolic changes in patients of severe acute respiratory syndrome (SARS) and SARS-related post-traumatic stress disorder (PTSD) and elucidate the relationship between PTSD, SARS and brain metabolism. METHODS A total of 58 convalescent SARS patients were evaluated by the scores of impact of event scale (IES), self-rating anxiety scale (SAS) and self-rating depression scale (SDS), etc. And 44 of them participated in (1)H-MRS study. There were 18 patients (M:F = 6:12) with SARS-related PTSD and 26 convalescent SARS patients without PTSD. And the procedures were repeated in 18 age, gender and education-matched normal control subjects. In all patients and controls, the regions of interest on magnetic resonance spectroscopy (MRS) were anterior cingulate gyrus (ACG), posterior cingulate gyrus (PCG), left anterior periventricular white matter (LAPWM) and left posterior periventricular white matter (LPPWM). And the correlations between MRS findings and the scores of SAS, SDS and IES were evaluated. RESULTS In comparisons with the control subjects (n = 18) and convalescent SARS patients without PTSD (n = 26), the SARS-related PTSD group (n = 18) had significantly lower N-acetylaspartate/creatine (NAA/Cr) in 4 regions of interest while the NAA/Cr of the convalescent SARS patients was significantly lower than controls in 3 regions of interest (ACG, PCG and LAPWM). The ratios of NAA/Cr showed no close correlation with the scores of IES, SAS and SDS in convalescent SARS patients. CONCLUSION The changes of brain metabolism in PTSD are caused by SARS. But their exact relationship awaits further explorations. SARS may also lead to the changes of brain metabolism.


title: Outbreak of histoplasmosis among travelers returning from El Salvador--Pennsylvania and Virginia, 2008.

text: Histoplasmosis is a fungal disease caused by infection with Histoplasma capsulatum. Histoplasmosis, which can be acquired from soil contaminated with bird or bat droppings, occurs worldwide and is one of the most common pulmonary and systemic mycoses in the United States. However, among international travelers returning from areas in which histoplasmosis is endemic, histoplasmosis is rare, accounting for <0.5% of all diseases diagnosed in this group. During February-March 2008, the Pennsylvania and Virginia departments of health investigated a cluster of respiratory illness among three mission groups that had traveled separately to El Salvador to renovate a church. This report summarizes the results of the investigation. Of 33 travelers in the three mission groups for whom information was available, 20 (61%) met the case definition for histoplasmosis. Persons who reported sweeping and cleaning outdoors (relative risk [RR] = 2.1, 95% confidence interval [CI] = 1.3-3.6), digging (RR = 2.6, CI = 1.1-6.1), or working in a bird or bat roosting area (RR = 1.8, CI = 1.3-2.4) had a greater risk for illness. The findings emphasize the need for travelers and persons involved in construction activities to use personal protective equipment and decrease dust-generation when working in areas where histoplasmosis is endemic. Clinicians should consider histoplasmosis as a possible cause of acute respiratory or influenza-like illness in travelers returning from areas in which histoplasmosis is endemic.


title: Role of nucleotides immediately flanking the transcription-regulating sequence core in coronavirus subgenomic mRNA synthesis.

text: The generation of subgenomic mRNAs in coronavirus involves a discontinuous mechanism of transcription by which the common leader sequence, derived from the genome 5' terminus, is fused to the 5' end of the mRNA coding sequence (body). Transcription-regulating sequences (TRSs) precede each gene and include a conserved core sequence (CS) surrounded by relatively variable sequences (5' TRS and 3' TRS). Regulation of transcription in coronaviruses has been studied by reverse-genetics analysis of the sequences immediately flanking a unique CS in the Transmissible gastroenteritis virus genome (CS-S2), located inside the S gene, that does not lead to detectable amounts of the corresponding mRNA, in spite of its canonical sequence. The transcriptional inactivity of CS-S2 was genome position independent. The presence of a canonical CS was not sufficient to drive transcription, but subgenomic synthesis requires a minimum base pairing between the leader TRS (TRS-L) and the complement of the body TRS (cTRS-B) provided by the CS and its adjacent nucleotides. A good correlation was observed between the free energy of TRS-L and cTRS-B duplex formation and the levels of subgenomic mRNA S2, demonstrating that base pairing between the leader and body beyond the CS is a determinant regulation factor in coronavirus transcription. In TRS mutants with increasing complementarity between TRS-L and cTRS-B, a tendency to reach a plateau in DeltaG values was observed, suggesting that a more precise definition of the TRS limits might be proposed, specifically that it consists of the central CS and around 4 nucleotides flanking 5' and 3' the CS. Sequences downstream of the CS exert a stronger influence on the template-switching decision according to a model of polymerase strand transfer and template switching during minus-strand synthesis.


Improving physician flow and efficiency in the emergency department.


title: Characterization of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike glycoprotein-mediated viral entry.

text: Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is a rapidly emerging pathogen with potentially serious consequences for public health. Here we describe conditions that result not only in the efficient expression of the SARS-CoV spike (S) protein on the surface of cells, but in its incorporation into lentiviral particles that can be used to transduce cells in an S glycoprotein-dependent manner. We found that although some primate cell lines, including Vero E6, 293T and Huh-7 cells, could be efficiently transduced by SARS-CoV S glycoprotein pseudoviruses, other cells lines were either resistant or very poorly permissive to virus entry. Infection by pseudovirions could be inhibited by several lysosomotropic agents, suggesting a requirement for acidification of endosomes for efficient S-mediated viral entry. In addition, we were able to develop a cell-cell fusion assay that could be used to monitor S glycoprotein-dependent membrane fusion. Although proteolysis did not enhance the infectivity of cell-free pseudovirions, trypsin activation is required for cell-cell fusion. Additionally, there was no apparent pH requirement for S glycoprotein-mediated cell-cell fusion. Together, these studies describe important tools that can be used to study SARS-CoV S glycoprotein structure and function, including approaches that can be used to identify inhibitors of the entry of SARS-CoV into target cells.


title: Insurance and epidemics: SARS, West Nile virus and Nipah virus.

text: Severe acute respiratory syndrome (SARS) reminds us that sudden disease emergence is a permanent part of our world--and should be anticipated in our planning. Historically the emergence of new diseases has had little or no impact beyond a small, localized cluster of infections. However, given just the right conditions, a highly virulent pathogen can suddenly spread across time and space with massive consequences, as has occurred on several occasions in human history. In the wake of the SARS outbreak, we are now forced to confront the unpleasant fact that human activities are increasing the frequency and severity of these kinds of emergences. The idea of more frequent biological "invasions" with economic and societal impacts comparable to SARS, presents stakeholders in and the global economy with unprecedented new risks, challenges and even opportunities. As a major contributor to economic stability, the insurance industry must follow these trends very closely and develop scenarios to anticipate these events.


title: Neurally adjusted ventilatory assist in patients with acute respiratory failure: study protocol for a randomized controlled trial.

text: BACKGROUND Patient-ventilator asynchrony is a common problem in mechanically ventilated patients with acute respiratory failure. It is assumed that asynchronies worsen lung function and prolong the duration of mechanical ventilation (MV). Neurally Adjusted Ventilatory Assist (NAVA) is a novel approach to MV based on neural respiratory center output that is able to trigger, cycle, and regulate the ventilatory cycle. We hypothesized that the use of NAVA compared to conventional lung-protective MV will result in a reduction of the duration of MV. It is further hypothesized that NAVA compared to conventional lung-protective MV will result in a decrease in the length of ICU and hospital stay, and mortality. METHODS/DESIGN This is a prospective, multicenter, randomized controlled trial in 306 mechanically ventilated patients with acute respiratory failure from several etiologies. Only patients ventilated for less than 5 days, and who are expected to require prolonged MV for an additional 72 h or more and are able to breathe spontaneously, will be considered for enrollment. Eligible patients will be randomly allocated to two ventilatory arms: (1) conventional lung-protective MV (n = 153) and conventional lung-protective MV with NAVA (n = 153). Primary outcome is the number of ventilator-free days, defined as days alive and free from MV at day 28 after endotracheal intubation. Secondary outcomes are total length of MV, and ICU and hospital mortality. DISCUSSION This is the first randomized clinical trial examining, on a multicenter scale, the beneficial effects of NAVA in reducing the dependency on MV of patients with acute respiratory failure. TRIAL REGISTRATION ClinicalTrials.gov website ( NCT01730794 ). Registered on 15 November 2012.


The top 10 hot topics in aging.


What's in this issue.


title: T cell antiviral effector function is not dependent on CXCL10 following murine coronavirus infection.

text: The chemokine CXCL10 is expressed within the CNS in response to intracerebral infection with mouse hepatitis virus (MHV). Blocking CXCL10 signaling results in increased mortality accompanied by reduced T cell infiltration and increased viral titers within the brain suggesting that CXCL10 functions in host defense by attracting T cells into the CNS. The present study was undertaken to extend our understanding of the functional role of CXCL10 in response to MHV infection given that CXCL10 signaling has been implicated in coordinating both effector T cell generation and trafficking. We show that MHV infection of CXCL10(+/+) or CXCL10(-/-) mice results in comparable levels of T cell activation and similar numbers of virus-specific CD4+ and CD8+ T cells. Subsequent analysis revealed no differences in T cell proliferation, IFN-gamma secretion by virus-specific T cells, or CD8+ T cell cytolytic activity. Analysis of chemokine receptor expression on CD4+ and CD8+ T cells obtained from MHV-immunized CXCL10(+/+) and CXCL10(-/-) mice revealed comparable levels of CXCR3 and CCR5, which are capable of responding to ligands CXCL10 and CCL5, respectively. Adoptive transfer of splenocytes acquired from MHV-immunized CXCL10(-/-) mice into MHV-infected RAG1(-/-) mice resulted in T cell infiltration into the CNS, reduced viral burden, and demyelination comparable to RAG1(-/-) recipients of immune CXCL10(+/+) splenocytes. Collectively, these data imply that CXCL10 functions primarily as a T cell chemoattractant and does not significantly influence T cell effector response following MHV infection.


Summaries for patients. Description of the 2003 severe acute respiratory syndrome outbreak in Hong Kong.


title: [Expression and purification of recombinant SARS coronavirus spike protein].

text: A novel coronavirus (SARS-coronavirus, SARS-CoV) was discovered in association with cases of severe acute respiratory syndrome (SARS) recently. The first step in coronavirus infection is binding of the viral spike protein to certain receptor on host cells. The spike protein is the main surface antigen of the coronavirus and there should be antibodies against spike protein in patients serum. Thus, to develop and expression protein fragment from spike protein gene are the purposes of this experiment. Partial spike gene fragments (751-1925 bp, 2005-3410 bp, 1-1925 bp and 32-3659 bp) and its intact gene were cloned into pET32 or pGEX vectors, and transformed into competent Escherichia coli BL21(DE3) (pLysS), respectively. 63, 78, 98, 160 and 164 kD fusion proteins were successfully expressed with amounts of 35%, 34%, 24%, 17% and 5% of total cell protein. The soluble parts of the cell crude extract were then partially purified by GST affinity chromatography.


title: Allergic contact dermatitis from formaldehyde textile resins.

text: Formaldehyde-based resins have been used to create permanent-press finishes on fabrics since the 1920s. These resins have been shown to be potent sensitizers in some patients, leading to allergic contact dermatitis. This review summarizes the history of formaldehyde textile resin use, the diagnosis and management of allergic contact dermatitis from these resins, and current regulation of formaldehyde resins in textiles.


title: [Study on the mimic epitopes screening program severe acute respiratory syndromes antigen with random phage peptide library].

text: OBJECTIVE To screen the severe acute respiratory syndromes (SARS) mimotopes with random phage peptide library and to investigate their immunogenicity. METHODS Using SARS sera as selective molecule, a 12 mer phage peptide library was biopanned and positive clones containing the mimic epitopes were selected. The immuno-characteriation of the epitopes were then investigated. RESULTS 2 positive clones that having specific affinity to SARS sera were obtained. The DNA sequencing data showed no homology between the sequences of the deduced amino acid of the two mimic antigen peptides and the sequence of SARS. CONCLUSION SARS mimotopes were obtained by phage peptide library screening. This method might provide a new approach for SARS therapy and vaccine development.


Human West Nile virus, France.


AIDS--acquired immune deficiency syndrome.


title: [Bovine coronavirus reproduction in the continuous cell cultures at a temperature of 34 degrees C].

text: Cultivation of bovine coronavirus at 34 degrees C showed the relationship of the degree of its reproduction to the cellular system. The possibility of obtaining the double yield of bovine coronavirus was demonstrated in MDBK and Vero cell cultures at 34 degrees C.


title: Selective replication of coronavirus genomes that express nucleocapsid protein.

text: The coronavirus nucleocapsid (N) protein is a structural protein that forms a ribonucleoprotein complex with genomic RNA. In addition to its structural role, it has been described as an RNA-binding protein that might be involved in coronavirus RNA synthesis. Here, we report a reverse genetic approach to elucidate the role of N in coronavirus replication and transcription. We found that human coronavirus 229E (HCoV-229E) vector RNAs that lack the N gene were greatly impaired in their ability to replicate, whereas the transcription of subgenomic mRNA from these vectors was easily detectable. In contrast, vector RNAs encoding a functional N protein were able to carry out both replication and transcription. Furthermore, modification of the transcription signal required for the synthesis of N protein mRNAs in the HCoV-229E genome resulted in the selective replication of genomes that are able to express the N protein. This genetic evidence leads us to conclude that at least one coronavirus structural protein, the N protein, is involved in coronavirus replication.


title: A new strategy in design of +RNA virus infectious clones enabling their stable propagation in E. coli.

text: Infectious clone methodology is a valuable tool of modern experimental virology. However, its use is often constrained by the instability of infectious clone constructs during propagation in E. coli. To circumvent this problem, we have devised a strategy that could be suitable for design of +RNA virus molecular clones in general. An infectious clone is assembled as "infectious DNA," and expression of problem regions present in the viral cDNA is prevented during propagation in E. coli by insertion of short introns. To demonstrate the feasibility of this approach, a highly unstable Japanese encephalitis flavivirus infectious clone has been successfully converted into a remarkably stable infectious DNA construct with the specific infectivity of 10(6) pfu/microg in cell culture. The proposed strategy may be useful in the design of self-amplifying gene therapy vectors and development of new immunization methodologies, and could facilitate creation of molecular repositories of existing viral vaccines.


The differential response to anti IL-6 treatment in COVID-19: the genetic counterpart.


title: Further analysis of the genome of porcine epidemic diarrhoea virus.

text: We report here the continued determination and analysis of the nucleotide sequence of both wild type (wt) and cell culture adapted (ca) porcine epidemic diarrhoea coronavirus (PEDV). These studies were undertaken with two objectives in mind: the identification of common and divergent features in the genomic sequences of wt and ca PEDV which can explain the differences in virulence of these isolates and the further exploration of the relationship of PEDV to other coronaviruses.


[Impact of the COVID-19 pandemic on heart failure hospitalizations].


title: Transmissible gastroenteritis.

text: Transmissible gastroenteritis (TGE) virus, a coronavirus, causes an acute infection of the small intestine of the pig. The disease is rarely fatal in adult animals but can cause extensive mortality in the neonate. Since its first description in Britain in the 1950s, the disease has been epizootic but recently it has become established on some farms and an antigenically related respiratory virus has become endemic in the national herd over the past two years. Piglet immunity to TGE, which relies on passive protection from milk, has been impossible to achieve with vaccines and research has aimed at understanding the nature of the interaction between virus and the pig. Following infection of the pregnant sow, antibody-producing cells migrate to the mammary gland where they secrete virus neutralising antibodies into the milk; prospective vaccines will need to stimulate a similar response. The location and number of antigenic sites on the virus particle associated with neutralisation have been established with monoclonal antibodies and the role of the other viral genes in pathogenesis and immunity is being studied with genetic engineering techniques.


title: New Small-Diameter Forceps for Thoracoscopic Surgery: Technical Adaptations and Initial Experiences.

text: In this report, we describe the use of new small-diameter forceps (Endo Relief forceps) for port-reduced thoracoscopic surgery. Forceps were designed with end that were the same size and shape as conventional 5-mm forceps, except that the diameter of the shaft was decreased to 2.4 mm. Endo Relief forceps were used for thoracoscopic surgery in 18 patients. We retrospectively compared the frequency of grasping error between conventional small-diameter forceps and Endo Relief group. The mean surgical time was 57.5 minutes (range, 45-75 minutes). There were no complications, no recurrences of pneumothorax and lung cancer, and no deaths after surgery. There were no intraoperative complications and no need for a second surgery to open additional ports. The frequency of grasping error was significantly lower in the Endo Relief group compared to the conventional small-diameter forceps group (0.17 ± 0.23 vs 1.33 ± 0.22; P = 0.022). Our experience thus far indicates that this technique has cosmetic benefit and is as effective as the classic 3-port technique for experienced thoracoscopic surgeons. Endo Relief forceps has a safety comparing conventional small-diameter forceps.


title: Papain-like protease (PLpro) inhibitory effects of cinnamic amides from Tribulus terrestris fruits.

text: Tribulus terrestris fruits are well known for their usage in pharmaceutical preparations and food supplements. The methanol extract of T. terrestris fruits showed potent inhibition against the papain-like protease (PLpro), an essential proteolylic enzyme for protection to pathogenic virus and bacteria. Subsequent bioactivity-guided fractionation of this extract led to six cinnamic amides (1-6) and ferulic acid (7). Compound 6 emerged as new compound possessing the very rare carbinolamide motif. These compounds (1-7) were evaluated for severe acute respiratory syndrome coronavirus (SARS-CoV) PLpro inhibitory activity to identify their potencies and kinetic behavior. Compounds (1-6) displayed significant inhibitory activity with IC50 values in the range 15.8-70.1 µM. The new cinnamic amide 6 was found to be most potent inhibitor with an IC50 of 15.8 µM. In kinetic studies, all inhibitors exhibited mixed type inhibition. Furthermore, the most active PLpro inhibitors (1-6) were proven to be present in the native fruits in high quantities by HPLC chromatogram and liquid chromatography with diode array detection and electrospray ionization mass spectrometry (LC-DAD-ESI/MS).


title: Monoclonal antibody-based immunohistochemical detection of porcine epidemic diarrhea virus antigen in formalin-fixed, paraffin-embedded intestinal tissues.

text: An immunohistochemistry technique was developed for the diagnosis of porcine epidemic diarrhea virus (PEDV). The technique was tested on formalin-fixed, paraffin-embedded intestinal tissues from piglets naturally infected with PEDV. Five different monoclonal antibodies (MAbs) were tested in this study. PEDV antigen was consistently detected in the PLP (4% paraformaldehyde, 100 mM L-lysine dihydrochloride, 10 mM sodium m-periodate in phosphate-buffered saline)-fixed PEDV-infected Vero cells or formalin-fixed, paraffin-embedded intestinal tissues from piglets naturally infected with PEDV. The C9-2-2 MAb gave the strongest reactivity and least background staining, detecting 10 of 10 infected pigs. The positive reaction was cytoplasmic. Positive enterocytes were distributed over the tip and along the sides of atrophied or fused villi in the jejunum and ileum. Positive-staining cells were not detected in the crypts. No staining was observed in cecum and colon. No positive cells were observed when the C9-2-2 MAb was reacted with the tissue sections from noninfected piglets or from transmissible gastroenteritus virus (TGEV)- and rotavirus-infected piglets. The selected anti-PEDV MAbs tested on formalin-fixed, paraffin-embedded tissue sections are useful for diagnosis when virus isolation is not available. This method would be of particular value in countries where both PEDV and TGEV are epizootic and would aid in differentiating between PEDV and TGEV infection.


title: Surge capacity mechanical ventilation.

text: Mechanical ventilation in a situation of mass casualty respiratory failure will require a substantial increase in the capacity for mechanical ventilation, to prevent unnecessary mortality. Concern over the difficulties of treating large numbers of patients with respiratory failure is exceeded only by our lack of experience on which to base decisions. This review evaluates the likely scenarios that could lead to mass casualty respiratory failure and the types of respiratory failure anticipated. A literature review was conducted, using the National Library of Medicine Medical Subject Headings terms "mass casualty respiratory failure," "pandemic flu," "disaster preparedness," and "mass casualty care." Papers were reviewed for relevance to the topic. There is little historical or empirical evidence upon which to base decisions regarding mass casualty respiratory failure and augmenting positive-pressure ventilation capacity. Matching the degree of respiratory impairment anticipated from the most likely mass casualty scenarios allows conclusions to be drawn regarding the performance characteristics of ventilators required for these situations. Little is known about the success of mechanical-ventilator stockpiling for mass casualty respiratory failure. Careful planning with an emphasis on matching ventilator performance to patient need and caregiver skill is critical to appropriate stockpile choices.


title: Evaluation of Pharmacokinetics, and Bioavailability of Higher Doses of Tocotrienols in Healthy Fed Humans.

text: BACKGROUND Tocotrienols has been known to lower serum lipid parameters below 500 mg/d, while increase lipid parameters at higher dose of 750 mg/d. δ-Tocotrienol has a novel inflammatory property of concentration-dependent inhibition and activation. Therefore, inhibition (anti-inflammatory) property of tocotrienols at low doses is useful for cardiovascular disease, whereas, activation (pro-inflammatory) property using high dose is found effective for treatments of various types of cancer. We have recently described plasma bioavailability of 125 mg/d, 250 mg/d and 500 mg/d doses of δ-tocotrienol in healthy fed subjects, which showed dose-dependent increases in area under the curve (AUC) and maximum concentration (Cmax). Hence, in the current study, higher doses of tocotrienols have used to analyze its effect on plasma pharmacokinetic parameters. AIMS To evaluate the safety and bioavailability of higher doses (750 mg and 1000 mg) of annatto-based tocotrienols in healthy fed subjects. All four isomers (α-, β-, γ-, δ-) of tocols (tocotrienols and tocopherols) present in the plasmas of subjects were quantified and analyzed for various pharmacokinetic parameters. STUDY DESIGN An open-label, randomized study was performed to analyze pharmacokinetics and bioavailability of δ-tocotrienol in 6 healthy fed subjects. All subjects (3/dose) were randomly assigned to one of each dose of 750 mg or 1000 mg. Blood samples were collected at 0, 1, 2, 4, 6, 8 h intervals and all isomers of α-,β-,γ-,δ-tocotrienols, and tocopherols in plasmas were quantified by HPLC. RESULTS Oral administration of 750 and 1000 mg/d of tocotrienols resulted in dose-dependent increases in plasmas (ng/ml) AUCt0-t8 6621, 7450; AUCt0-∞ 8688, 9633; AUMC t0-∞ 52497, 57199; MRT 6.04, 5.93; Cmax 1444, 1592 (P<0.05), respectively, of δ-tocotrienol isomer. Moreover, both doses also resulted in plasmas Tmax 3.33-4 h; elimination half-life (t1/2 h) 2.74, 2.68; time of clearance (Cl-T, l/h) 0.086, 0.078; volume of distribution (Vd/f, mg/h) 0.34, 0.30; and elimination rate constant (ke; h-1) 0.25, 0.17, respectively of δ- tocotrienol isomer. Similar results of these parameters were reported for γ-tocotrienol, β- tocotrienol, α-tocotrienol, δ-tocopherol, γ-tocopherol, and β-tocopherol, except for α- tocopherol. CONCLUSIONS This study has described pharmacokinetics using higher doses of 750 mg/d and 1000 mg/d of δ-tocotrienol. These results confirmed earlier findings that Tmax was 3-4 h for all isomers of tocotrienols and tocopherols except for α-tocopherol (6 h). These higher doses of tocotrienols were found safe in humans and may be useful for treatments of various types of cancer, diabetes, and Alzheimer's disease.


Separation of anxiety and depressive disorders: blind alley in psychopharmacology and classification of disease.


title: BILATERAL HYDRONEPHROSIS IN A SUGAR GLIDER (PETAURUS BREVICEPS).

text: An adult, intact male sugar glider ( Petaurus breviceps ) presented for acute caudal abdominal swelling. Treatment by the referring veterinarian included aspiration of urine from the swelling. On physical examination, mild depression, pale mucus membranes, and caudal abdominal swelling were noted. Focused ultrasonographic assessment revealed a fluid-filled caudal abdominal structure and subjective bladder wall thickening. The following day, the sugar glider was severely depressed. Hematology results included hypoglycemia, hyperkalemia, hyponatremia, and azotemia. Ultrasonography revealed bilateral hydronephrosis and hydroureter. Despite supportive care, the animal died. Postmortem examination confirmed bilateral ureteral dilation, renal petechial hemorrhage, and dilation of the right renal pelvis. Submucosal edema, hemorrhage, and lymphoplasmacytic infiltration of the urinary bladder, ureters, and renal pelvises were noted. Hyperplasia of the urinary bladder and ureteral epithelium, coupled with inflammation, may have caused functional obstruction leading to bilateral hydronephrosis and hydroureter. This is the first reported case of hydronephrosis in a marsupial.


title: [Detection of cell apoptosis in the pathological tissues of patients with SARS and its significance].

text: OBJECTIVE To explore the role and mechanism of apoptosis in severe acute respiratory syndrome (SARS). METHODS Klenow-FragELTM DNA Fragmentation Detection Kit and immunohistochemical alkaline phosphatase detection reagent kit were used to detect cell apoptosis and expressions of CD68, CD20, CD4, CD8 and CD45RA in the pathological tissues of SARS patients. RESULTS Apoptotic cells increased significantly in the spleen, lung and lymph nodes of SARS patients as compared with normal tissues. The apoptotic cells included pneumocytes, lymphocytes and monocytes, and CD68+ monocytes were observed in abundance in the lung, spleen and lymph nodes of SARS patients. In the lung tissue of the patients, few CD20+/CD45RA+ B cells and CD4+/CD8+ T cells were spotted, and CD20+/CD45RA+ B cells along with CD4+/CD8+ T cells were also significantly decreased in the spleen and lymph nodes, where few conserved B and T cells underwent apoptosis. CONCLUSIONS Apoptosis is a general phenomenon in SARS, and the invasive cells in the pathological tissues are primarily monocytes, suggesting that apoptosis and invasion of monocyte play important roles in the progression of SARS. The cell apoptosis and decreased number of T cell and B cells in the lungs and CD4+/CD8+ T cells and CD20+/CD45RA+ B cells in the spleen and lymph nodes indicate that the SARS virus may exercise immune cell-killing effect to some extent during its pathogenesis.


title: Posttraumatic Stress Disorder Among Older Patients with Chronic Obstructive Pulmonary Disease.

text: This study explored (1) the incidence of posttraumatic stress disorder (PTSD) resulting from past trauma among older patients with COPD and (2) whether PTSD and COPD severity would relate to psychiatric co-morbidity and health-related quality of life. Eighty-five older patients completed the Hospital Anxiety and Depression Scale, the Chronic Respiratory Questionnaire, the Posttraumatic Stress Diagnostic Scale and the Medical Outcomes Short Form 12. The results showed that 55, 39 and 6 % had no, partial and full-PTSD respectively. Partial least squares showed that PTSD was significantly correlated with COPD severity which in turn was significantly correlated with health-related quality of life and psychiatric co-morbidity. Mediational analysis showed that the emotional symptoms of COPD mediated between PTSD and the mental health functioning of health-related quality of life and between PTSD and depression. To conclude, PTSD from past trauma was related to the severity of COPD for older patients. In particular, it impacted on the elevated emotional arousal of COPD severity. In turn, COPD severity impacted on older patients' general psychological well-being and depression.