title: A single immunization with a rhabdovirus-based vector expressing severe acute respiratory syndrome coronavirus (SARS-CoV) S protein results in the production of high levels of SARS-CoV-neutralizing antibodies.

text: Foreign viral proteins expressed by rabies virus (RV) have been shown to induce potent humoral and cellular immune responses in immunized animals. In addition, highly attenuated and, therefore, very safe RV-based vectors have been constructed. Here, an RV-based vaccine vehicle was utilized as a novel vaccine against severe acute respiratory syndrome coronavirus (SARS-CoV). For this approach, the SARS-CoV nucleocapsid protein (N) or envelope spike protein (S) genes were cloned between the RV glycoprotein G and polymerase L genes. Recombinant vectors expressing SARS-CoV N or S protein were recovered and their immunogenicity was studied in mice. A single inoculation with the RV-based vaccine expressing SARS-CoV S protein induced a strong SARS-CoV-neutralizing antibody response. The ability of the RV-SARS-CoV S vector to confer immunity after a single inoculation makes this live vaccine a promising candidate for eradication of SARS-CoV in animal reservoirs, thereby reducing the risk of transmitting the infection to humans.


Strange days.


title: Eradication versus control: the economics of global infectious disease policies.

text: A disease is controlled if, by means of a public policy, the circulation of an infectious agent is restricted below the level that would be sustained by individuals acting independently to control the disease. A disease is eliminated if it is controlled sufficiently to prevent an epidemic from occurring in a given geographical area. Control and elimination are achieved locally, but a disease can only be eradicated if it is eliminated everywhere. Eradication is plainly a more demanding goal, but it has two advantages over control. First, the economics of eradication can be very favourable when eradication not only reduces infections but also avoids the need for vaccinations in future. Indeed, when eradication is feasible, it will either pay to control it to a fairly low level or to eradicate it. This suggests that, from an economics perspective, diseases that are eliminated in high-income countries are prime candidates for future eradication efforts. Second, the incentives for countries to participate in an eradication initiative can be strong; indeed they can be even stronger than an international control programme. Moreover, high-income countries typically benefit so much that they will be willing to finance elimination in developing countries. Full financing of an eradication effort by nation-states is not always guaranteed, but it can be facilitated by a variety of means. Hence, from the perspective of economics and international relations, eradication has a number of advantages over control. The implications for smallpox and polio eradication programmes are discussed.


title: Safety and Efficacy of a Novel Double-Stapling Technique for Distal Esophageal Resection and Esophago-Jejunal Anastomosis.

text: BACKGROUND The purpose of this study was to evaluate safety and efficacy of a new esophagojejunal anastomosis (EJA) technique allowing the insertion of the anvil of a common circular stapler without hand-sewn securing. METHODS From August 2014 to May 2015, 20 consecutive patients with esophagogastric junction adenocarcinoma underwent surgery. EJA was performed using a new technique; the free margins of the esophageal stump were suspended and the anvil of a circular stapler on a new dedicated and registered support bar (characterized by a push-rod making possible to hook-unhook the anvil of the circular stapler) was inserted into the lumen. Subsequently, the linear suturing stapler was closed over the bar and fired to suture the distal stump of the esophagus; the bar was retracted and the push-rod of the anvil was pulled out through the linear suture. Finally, the anastomosis was performed using a circular stapler. RESULTS There were no intraoperative complications, and R0 resection was achieved in all cases. Postoperative course has been uneventful for 18 patients (90%). Only 1 patient (5%) developed fistula, conservatively treated. CONCLUSIONS Our preliminary clinical experience suggested that this technique was safe and efficient (for all online suppl. material, see www.karger.com/doi/10.1159/000446856).


China's Coronavirus-Induced Paralysis Threatens U.S. Drug Supply Chain.


title: A novel astrovirus species in the gut of yaks with diarrhoea in the Qinghai–Tibetan Plateau, 2013.

text: The yak (Bos grunniens) is an iconic symbol in the high-altitude region of the Qinghai–Tibetan Plateau. Diarrhoea is a common disease in yaks, resulting in major economic losses. To investigate the diversity of viral species, we reported the metagenomics-derived virome in a pooled faecal sample of 20 diarrhoeic yaks. The nine viruses found in the pooled diarrhoeic samples, in order of abundance of nucleic acid sequence, were influenza A virus, bovine viral diarrhoea virus (BVDV), rotavirus, ungulate tetraparvovirus 1 (bovine hokovirus), astrovirus (AstV), bovine enterovirus, hepatitis E virus, kobuvirus and woodchuck hepatitis virus. Compared with healthy yaks, only AstV had a significantly higher prevalence rate in diarrhoeal samples, indicating a correlation with the clinical symptoms of diarrhoea in yaks. To further investigate the molecular characterization of yak AstV, a near-full genome was obtained from a diarrhoeic sample. It was 6243 bp in length and shared 46.4–66.2 % similarity with other related bovine AstVs from faeces. Phylogenetic analysis of the genome demonstrated that the yak AstV fell within the bovine AstVs cluster, but was located in a unique lineage, suggesting a novel AstV species was identified in yaks. Interestingly, the ORF2 region of yak AstV had closer similarity and genetically relationship with deer AstV strain CcAstV-2 than that of the bovine AstVs. Further analysis showed that one possible interspecies recombination event occurred in ORF2. In summary, this study expanded our understanding of the viral communities of diarrhoeal yaks and identified a novel AstV that was associated with diarrhoea in yaks.


title: Mucosal and systemic antibody responses to bovine coronavirus structural proteins in experimentally challenge-exposed calves fed low or high amounts of colostral antibodies.

text: Ten colostrum-deprived calves were assigned to 1 of 2 treatment groups (5 calves/group), and fed colostrum that had either low (naturally infected cows) or high (immunized cows) antibody titers to bovine coronavirus (BCV). All calves were inoculated orally and intranasally with virulent BCV when they were 24 to 48 hours old and challenge exposed 21 days later. Blood, feces, nasal secretions, tears, saliva, and bronchoalveolar lavage (BAL) fluids were collected weekly from each calf for 5 weeks after inoculation. The titers to whole BCV or the relative amounts of isotype-specific antibodies to BCV structural proteins were evaluated in these samples by ELISA or immunoblotting, respectively. Both pools of colostrum contained primarily IgG1, IgG2, and IgA antibodies to the E2 and E3 BCV proteins. Calves fed the high-titer colostrum had correspondingly higher amounts of passive IgG1 and IgA antibodies to whole BCV and to the E2 and E3 BCV proteins in serum, feces, and BAL fluid at postinoculation week 1 than those calves fed low-titer colostrum. Active IgG1, IgA, and IgM antibody responses in serum and active IgA and IgM antibody responses in most mucosal secretions to whole BCV and to the E2 and E3 proteins were lower or delayed in calves fed high-titer colostrum, compared with responses in calves fed low-titer colostrum. In contrast, increased responses to the BCV N protein were observed in all samples (except in serum and BAL fluid) in the calves fed high-titer colostrum, compared with calves fed low-titer colostrum. Upon challenge exposure, responses to E2 and E3 BCV proteins in serum and BAL fluid were lower in the group fed high-titer colostrum, compared with those in the group fed low-titer colostrum. Our findings indicate that the level of passive immunity in calves at the time of BCV inoculation can influence the development of active antibody responses in serum, feces, and mucosal secretions to whole BCV and to some BCV proteins individually.


title: Early detection of acute rhinovirus infections by a rapid reverse transcription-PCR assay.

text: The development of a rhinovirus (RV)-RNA-specific reverse transcription (RT)-PCR assay is complicated by the close homology between the RV and enterovirus (EV) genomes in the highly conserved 5'-noncoding region, which is chosen for primer design in most RT-PCR assays. We have developed a sensitive, rapid, and RV-specific nested RT-PCR assay and have used it to test nasopharyngeal aspirates from 556 patients presenting with acute respiratory tract infections. RV RNA was detected by nested RT-PCR not only in all of 52 samples that were RV positive by virus isolation methods but also in 124 of 367 samples that were negative by virus isolation methods and enzyme-linked immunosorbent assay (ELISA). In addition, in 23 of 137 samples that were positive for a different respiratory virus by virus isolation and/or ELISA, RV RNA was detected by RT-PCR. EVs, adenoviruses, respiratory syncytial viruses, coronaviruses, and influenza and parainfluenza viruses, including clinical isolates as well as stock viruses, were not amplified in our RV-specific RT-PCR assay, indicating that this assay was highly specific. The processing time was less than 2 days for the RT-PCR, as opposed to up to 2 weeks for virus isolation. These results indicate that nested RT-PCR is more sensitive than conventional methods for the detection of RV in patients experiencing acute respiratory tract infections and represents the only reliable tool for the early laboratory diagnosis of RV infections. This is especially important in light of new opportunities for therapy currently being developed.


title: A retrospective study of 78 patients with severe acute respiratory syndrome.

text: OBJECTIVE To summarize the clinical features of severe acute respiratory syndrome (SARS) and to discuss diagnosis and management of the disease. METHODS A retrospective study was conducted on 78 cases of SARS referred to the Guangzhou Institute of Respiratory Diseases(GIRD) between December 22, 2002 and near the end of March 2003. Items reviewed cover all data concerning clinical manifestations, laboratory investigation and radiology. RESULTS The patients on the study consisted of 42 males and 36 females, aged 20-75 yrs (mean age 37.5 +/- 11.6 yrs), including 44 affected health-care professionals. Clinical symptoms seen in the group were fever (100.0%), cough (88.5%), and dyspnea (79.5%). There were 12 cases (15.3%) with WBCs < 4.0 x 10(9) /L, 49 cases (62.8%) ranging between (4.0 -10.0) x 10(9) /L and 17 cases (21.8%) over 10.0 x 10(9) /L. The average was (7.58 +/- 4.96) x 10(9) /L, with 0.75 +/- 0.14 (neutrophil) and 0.18 +/- 0.11 (lymphocyte). Chest films and CT scanning revealed changes related to pneumonia. The transmission of the disease was likely via close contact with contagious droplets. The prevalences of acute lung injury (ALI, in 37 cases) and acute respiratory distress syndrome (ARDS, 21 out of 37 cases) were considerably high among the patients. Seven patients who developed ARDS complicated with multiple organs dysfunction syndrome (MODS) died. CONCLUSIONS A history of close contact, fever, sign of pneumonia by X-ray and normal-to-lowered WBC counts are favorable for the diagnosis of SARS. Recognition of ALI as the important index for critical SARS and comprehensive supportive management are of paramount in decreasing the mortality of the disease.


Bovine coronavirus-induced cytopathic expression and plaque formation: host cell and virus strain determine trypsin dependence.


1646: UTILIZING ECMO AS LIFESAVING INTERVENTION IN A PATIENT WITH THROMBOSED MECHANICAL MITRAL VALVE.


title: Transmissible gastroenteritis coronavirus carrier sow.

text: A sow infected with virulent transmissible gastroenteritis virus (TGEV) shed virulent virus in her feces for 18 months. The virus was isolated from rectal swabs beginning 2 days postexposure (PE) and continued at irregular intervals. Virus shedding was detected on 24 separate occasions. The titer of the virus shed ranged from < 1 x 10(2) pfu/ml to 7.2 x 10(3) pfu/ml, while the duration of the shedding ranged from 1 to 5 consecutive days. Inoculation of 3-day-old piglets with TGEV isolated from the sow proved the virus was virulent throughout the study. Virulent TGEV was isolated from the spleen, mesenteric lymph nodes, and the liver of the sow 544 days PE. This study demonstrates an apparently healthy sow can be a reservoir and shed virulent TGEV for an extended period of time.


title: Histidine at residue 99 and the transmembrane region of the precursor membrane prM protein are important for the prM-E heterodimeric complex formation of Japanese encephalitis virus.

text: The formation of the flavivirus prM-E complex is an important step for the biogenesis of immature virions, which is followed by a subsequent cleavage of prM to M protein through cellular protease to result in the production and release of mature virions. In this study, the intracellular formation of the prM-E complex of Japanese encephalitis virus was investigated by baculovirus coexpression of prM and E in trans in Sf9 insect cells as analyzed by anti-E antibody immunoprecipitation and sucrose gradient sedimentation analysis. A series of carboxyl-terminally truncated prM mutant baculoviruses was constructed to demonstrate that the truncations of the transmembrane (TM) region resulted in a reduction of the formation of the stable prM-E complex by approximately 40% for the TM1 (at residues 130 to 147 [prM130-147]) truncation and 20% for TM2 (at prM153-167) truncation. Alanine-scanning site-directed mutagenesis on the prM99-103 region indicated that the His99 residue was the critical prM binding element for stable prM-E heterodimeric complex formation. The single amino acid mutation at the His99 residue of prM abolishing the prM-E interaction was not due to reduced expression or different subcellular location of the mutant prM protein involved in prM-E interactions as characterized by pulse-chase labeling and confocal scanning microscopic analysis. Recombinant subviral particles were detected in the Sf9 cell culture supernatants by baculovirus coexpression of prM and E proteins but not with the prM H99A mutant. Sequence alignment analysis was further conducted with different groups of flaviviruses to show that the prM H99 residues are generally conserved. Our findings are the first report to characterize the minimum binding elements of the prM protein that are involved in prM-E interactions of flaviviruses. This information, concerning a molecular framework for the prM protein, is considered to elucidate the structure/function relationship of the prM-E complex synthesis and provide the proper trajectory for flavivirus assembly and maturation.


title: Further observations on coronavirus infection of primate CNS.

text: Previously, we demonstrated that intracerebral (IC) inoculation of a murine coronavirus, MHV-JHM, into two species of primates can result in acute encephalomyelitis (Murray et al., 1992a). Infectious virus isolated from acutely infected animals, designated JHM-OMp1, was inoculated IC into a second group of monkeys. In this report we describe observations on the acutely infected animals and those surviving the acute infection were sacrificed at later times post-infection. Results from dual in situ hybridization/immunohistochemistry screening of tissues show that astrocytes are target cells in white matter lesions during acute infection. In animals sacrificed 150 days post-infection, areas of demyelinated gliotic lesions, prominent in the spinal cord, were seen throughout the neuraxis. No virus products were detected in these late-infection lesions.


title: Controlling subcellular delivery to optimize therapeutic effect.

text: This article focuses on drug targeting to specific cellular organelles for therapeutic purposes. Drugs can be delivered to all major organelles of the cell (cytosol, endosome/lysosome, nucleus, nucleolus, mitochondria, endoplasmic reticulum, Golgi apparatus, peroxisomes and proteasomes) where they exert specific effects in those particular subcellular compartments. Delivery can be achieved by chemical (e.g., polymeric) or biological (e.g., signal sequences) means. Unidirectional targeting to individual organelles has proven to be immensely successful for drug therapy. Newer technologies that accommodate multiple signals (e.g., protein switch and virus-like delivery systems) mimic nature and allow for a more sophisticated approach to drug delivery. Harnessing different methods of targeting multiple organelles in a cell will lead to better drug delivery and improvements in disease therapy.


Coronaviruses: a historical perspective.


title: [Dynamic inflammatory response in a critically ill COVID-19 patient treated with corticosteroids].

text: OBJECTIVE To investigate the effect of corticosteroids therapy on the inflammatory response in a critically ill coronavirus disease 2019 (COVID-19) patient. METHODS A 55-year old female patient with critical ill COVID-19 was admitted in Taizhou Hospital on January 19, 2020. The patient was treated with methylprednisolone 80 mg on the 2nd day after admission. Thereafter, the dose was adjusted in a timely manner and the therapy lasted for 13 days. The peripheral lymphocyte subsets (CD3+T, CD4+ T, CD8+ T, NK cells, B cells), as well as serum levels of lymphocyte factors (IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ) were dynamically monitored. RESULTS On D1 of admission, the numbers of peripheral blood CD3+ T, CD4+ T, CD8+ T, and NK cells were significantly lower than the normal range. With the improvement of the disease, the numbers of CD3+ T, CD8+ T and CD4 + T cells gradually recovered and showed a linear growth trend (linear fitting equation: Y=18.59X+109.4, P<0.05). On D2 of admission, the patient's IL-6 and IL-10 levels were significantly higher than normal values, IFN-γ was at a normal high value, and then rapidly decreased; IL-2, IL-4, and TNF-α were all in the normal range. On the D6 and D7, the IL-6 and IL-10 decreased to the normal range for the first time. On the D18, the sputum virus nucleic acid test was negative for the first time, and the fecal virus nucleic acid test was still positive; on the D20 the sputum and fecal virus nucleic acid test were both negative. On D34, the patient recovered and was discharged. At the discharge the muscle strength score of the patient was 44 and the daily life ability evaluation was 90. CONCLUSIONS In the absence of effective antiviral drugs, early use of appropriate doses of corticosteroids in critically ill patient with COVID-19 can quickly alleviate inflammatory response and improve clinical symptoms, however, it may reduce the number of T cells, and to adjust the dose in time is necessary.


title: Posterior interosseous neuropathy: Supinator syndrome vs fascicular radial neuropathy.

text: OBJECTIVE To investigate the spatial pattern of lesion dispersion in posterior interosseous neuropathy syndrome (PINS) by high-resolution magnetic resonance neurography. METHODS This prospective study was approved by the local ethics committee and written informed consent was obtained from all patients. In 19 patients with PINS and 20 healthy controls, a standardized magnetic resonance neurography protocol at 3-tesla was performed with coverage of the upper arm and elbow (T2-weighted fat-saturated: echo time/repetition time 52/7,020 milliseconds, in-plane resolution 0.27 × 0.27 mm2). Lesion classification of the radial nerve trunk and its deep branch (which becomes the posterior interosseous nerve) was performed by visual rating and additional quantitative analysis of normalized T2 signal of radial nerve voxels. RESULTS Of 19 patients with PINS, only 3 (16%) had a focal neuropathy at the entry of the radial nerve deep branch into the supinator muscle at elbow/forearm level. The other 16 (84%) had proximal radial nerve lesions at the upper arm level with a predominant lesion focus 8.3 ± 4.6 cm proximal to the humeroradial joint. Most of these lesions (75%) followed a specific somatotopic pattern, involving only those fascicles that would form the posterior interosseous nerve more distally. CONCLUSIONS PINS is not necessarily caused by focal compression at the supinator muscle but is instead frequently a consequence of partial fascicular lesions of the radial nerve trunk at the upper arm level. Neuroimaging should be considered as a complementary diagnostic method in PINS.


title: Critical role for arginine methylation in adenovirus-infected cells.

text: During the late stages of adenovirus infection, the 100K protein (100K) inhibits the translation of cellular messages in the cytoplasm and regulates hexon trimerization and assembly in the nucleus. However, it is not known how it switches between these two functions. Here we show that 100K is methylated on arginine residues at its C terminus during infection and that this region is necessary for binding PRMT1 methylase. Methylated 100K is exclusively nuclear. Mutation of the third RGG motif (amino acids 741 to 743) prevents localization to the nucleus during infection, suggesting that methylation of that sequence is important for 100K shuttling. Treatment of infected cells with methylation inhibitors inhibits expression of late structural proteins. These data suggest that arginine methylation of 100K is necessary for its localization to the nucleus and is a critical cellular function necessary for productive adenovirus infection.


title: [Effect of SARS-associated coronavirus on peripheral blood picture and liver function].

text: OBJECTIVE To study peripheral blood picture and liver function of severe acute respiratory syndrome (SARS), and to accumulate more experience in prevention and treatment this syndrome. METHODS The peripheral blood picture and liver function of 169 cases with SARS were analyzed. These patients were admitted to five hospitals in Taiyuan from March to May, 2003. RESULTS Above 90.00% patients had normal or low white blood cell (WBC) count on the first day. From then on, during the second to third week, about 40.00% of the patients showed high WBC count average: (13.42+/-2.87)x10(9)/L at 15 th day and (13.68+/-3.26)x10(9)/L at 21 st day) and neutrophil (0.809+/-0.063 at 15 th day and 0.805+/-0.061 at 21 st day), and 21.97%-52.15% cases with low lymphocyte count (0.111+/-0.044-0.134+/-0.040). About 4.44%-12.76% of patients developed low platelet and 4.20%-33.33% of cases with low hemoglobin level. And 32.76%-62.50% of cases were found to have high alanine aminotransferase (above 40 U/L), 13.04%-40.00% cases with high aspartate aminotransferase (above 40 U/L) and 40.35%-72.00% cases with low albumin content. But serum total protein remained normal during the whole clinical course. CONCLUSION SARS cases may develop leucocytosis, low lymphocyte count, low platelet count, low hemoglobin level and abnormal liver function.


title: Increasing Role of Image-Guided Ablation in the Treatment of Musculoskeletal Tumors.

text: In the last decade, percutaneous treatment of musculoskeletal (MSK) tumors has become more established in routine clinical care while also undergoing a number of advancements. Ablative techniques to palliate painful skeletal metastases have gained wide acceptance, while goals for ablation have evolved to include local control of oligometastases and desmoid tumors. Bone consolidation or augmentation is now frequently used in conjunction with or instead of ablation of skeletal tumors to stabilize pathologic fractures or prevent further morbidity that could result from fractures caused by tumor progression. These procedures have traditionally been performed with cement injection, although additional percutaneous consolidation or stabilization devices have been developed. Techniques to monitor the ablation zone and adjacent structures intraprocedurally are now applied to increase the number of tumors amenable to treatment. These include methods to depict, displace, or monitor critical structures adjacent to targeted MSK tumors. Finally, the role of ablation in the comprehensive care of patients with MSK tumors continues to change with the evolving triage of patients between radiation therapy, surgical resection and stabilization, and percutaneous ablative and consolidative management.


title: Effects of inspiratory rise time on triggering work load during pressure-support ventilation: a lung model study.

text: BACKGROUND The rise in inspiratory flow is important during patient-triggered ventilation. Many ventilators incorporate a function to control the time to reach the targeted airway pressure (inspiratory rise time). However, it has not been clarified how inspiratory rise time affects inspiratory work load under various ventilator settings. In a bench study we investigated the effect of inspiratory rise time on inspiratory work load during pressure-support ventilation (PSV). METHODS We studied 6 ICU ventilators. We measured flow and pressure at the airway opening (P(ao)) at PEEP of 5 cm H(2)O, pressure-support of 5 cm H(2)O and 10 cm H(2)O, 4 triggering sensitivities, and inspiratory drives 300 mL, 500 mL, and 700 mL. The inspiratory-rise-time setting was not consistent between the ventilators, and we chose 3 inspiratory-rise-time levels with each ventilator. The inspiratory delay time (DT) was defined as the time between the onset of inspiration and the return of P(ao) to baseline, and was divided into 2 parts at the point of the lowest P(ao): before the lowest P(ao) (DT(1)), and after the lowest P(ao) (DT(2)). As an indicator of inspiratory work load we calculated the pressure-time-product (PTP) of the P(ao) over the DT. PTP was also divided into PTP(1) and PTP(2), at the point of the lowest P(ao). RESULTS Short inspiratory rise time reduced DT(2), PTP(1), and PTP(2), regardless of the pressure-support level, triggering sensitivity, or inspiratory drive. However, the inspiratory-rise-time setting did not affect DT(1). The PTP(1), PTP(2), and DT(2) values differed significantly among the ventilators. A combination of short inspiratory rise time, high PSV, and sharp triggering sensitivity resulted in the smallest PTP and DT values. CONCLUSIONS Short inspiratory rise time decreased inspiratory work load, regardless of the pressure-support level, triggering sensitivity, or inspiratory drive. Inspiratory work load can be maximally lowered by a combination of a short inspiratory rise time, a sharp triggering sensitivity, and a high inspiratory pressure-support level for a given patient's inspiratory effort.


title: The critical role of IFN-gamma in experimental coronavirus retinopathy.

text: PURPOSE Experimental coronavirus retinopathy (ECOR) is an animal model of progressive retinal disease that is first manifest as an acute retinal inflammation followed by chronic, immune-associated retinal degeneration in genetically susceptible, BALB/c mice. In retinal degeneration-resistant CD-1 mice, only the acute infection is seen. In the present study, interferon (IFN)-gamma production during ECOR was studied and its role evaluated in the clearance of infectious virus from the retina. METHODS BALB/c, CD-1, and IFN-gamma-deficient (IFN-gamma gko) mice were inoculated with the JHM strain of murine coronavirus by the intravitreal route. Mouse eyes were evaluated for infiltrating cells and major histocompatibility complex (MHC) expression by immunocytochemical staining. Isolated retinas were analyzed for IFN-gamma mRNA by RT-PCR, and sera were evaluated for IFN-gamma protein by ELISA assays. RESULTS Virus infection in BALB/c mice was associated with an increase in the incidence and levels of systemic IFN-gamma. Moreover, IFN-gamma mRNA was detected within the retinas of infected animals during the acute phase of the disease but was not detected in normal or mock-injected animals. IFN-gamma mRNA was detected at the time of T-cell infiltration, and earlier studies have shown that this is temporally related to granzyme B gene expression and the clearance of infectious virus from the retina. Retinal IFN-gamma mRNA was also associated with the upregulation of MHC class I and II molecules within the retina. When this infection occurred in IFN-gamma gko mice, the virus was unchecked, and the infection led to death. CONCLUSIONS These studies indicate that generation of IFN-gamma by cells infiltrating the retina is an essential part of an immune mechanism responsible for noncytolytic clearance of infectious virus from the retina.


title: Evidence of person-to-person transmission within a family cluster of novel coronavirus infections, United Kingdom, February 2013.

text: In February 2013, novel coronavirus (nCoV) infection was diagnosed in an adult male in the United Kingdom with severe respiratory illness, who had travelled to Pakistan and Saudi Arabia 10 days before symptom onset. Contact tracing identified two secondary cases among family members without recent travel: one developed severe respiratory illness and died, the other an influenza-like illness. No other severe cases were identified or nCoV detected in respiratory samples among 135 contacts followed for 10 days.


title: How did general practitioners protect themselves, their family, and staff during the SARS epidemic in Hong Kong?

text: CONTEXT Severe acute respiratory syndrome (SARS) is a newly emerging infectious disease and how the frontline community doctors respond to it is not known. OBJECTIVES To explore the impact of SARS on general practitioners (GPs) in Hong Kong. DESIGN A cross sectional survey. SETTING Community based primary care clinics. PARTICIPANTS 183 family medicine tutors affiliated with a local university. Postal survey sent to all tutors with a 74.8% response rate. MAIN OUTCOME MEASURES Change of clinical behaviour and practices during the epidemic; anxiety level of primary care doctors. RESULTS All agreed SARS had changed their clinical practices. Significant anxiety was found in family doctors. Three quarters of respondents recalled requesting more investigations while a quarter believed they had over-prescribed antibiotics. GPs who were exposed to SARS or who had worked in high infection districts were less likely to quarantine themselves (10.8% versus 33.3%; p<0.01; 6.5% versus 27.5%; p<0.01 respectively). Exposure to SARS, the infection rates in their working district, and anxiety levels had significant impact on the level of protection or prescribing behaviour. CONCLUSION The clinical practice of GPs changed significantly as a result of SARS. Yet, those did not quarantine themselves suggesting other factors may have some part to play. As failure to apply isolation precautions to suspected cases of SARS was one major reason for its spread, a contingency plan from the government to support family doctors is of utmost importance. Interface between private and public sectors are needed in Hong Kong to prepare for any future epidemics.


title: Update: severe acute respiratory syndrome--United States, 2003.

text: CDC, in collaboration with state and local health departments, the World Health Organization (WHO), and other partners, continues to investigate cases of severe acute respiratory syndrome (SARS). During November 1, 2002-May 7, 2003, a total of 6,903 SARS cases were reported to WHO from 29 countries, including the United States; 495 deaths (case-fatality proportion: 7.2%) have been reported. This report updates information on reported U.S. SARS cases.


title: Influential disease foci in epidemics and underlying mechanisms: a field experiment and simulations.

text: Pathogen invasions pose a growing threat to ecosystem stability and public health. Guidelines for the timing and spatial extent of control measures for pathogen invasions are currently limited, however. We conducted a field experiment using wheat (Triticum aestivum) stripe rust, caused by the wind-dispersed fungus Puccinia striiformis, to study the extent to which host heterogeneity in an initial outbreak focus influences subsequent disease spread. We varied the frequency of susceptible host plants in an initial outbreak focus and in the non-focus of experimental plots, and observed the progress of epidemics produced by artificial inoculation. The frequency of susceptible hosts in the initial outbreak focus increased the spread of stripe rust in the experimental plots, while frequency of susceptible hosts outside the initial outbreak focus did not. This suggests that factors influencing pathogen reproduction in the initial outbreak focus are key to the control of epidemics of stripe rust. Two mechanisms may underlie the field results. The first is the continuing, direct infection of susceptible hosts in areas outside the initial outbreak focus by disease propagules arriving from the initial outbreak focus. The second is highly local proliferation of disease caused by direct descendants of colonizing individuals originating from the initial outbreak focus. We considered these two alternatives in simulations of a generalized pathogen exhibiting fat-tailed dispersal, similar to P. striiformis. Simulations showed a dominant effect of conditions in the initial outbreak focus, in agreement with the field experiment, but indicated that, over time, this dominance may erode. Analysis of the duration of focal dominance led to the conclusion that both mechanisms contribute to the phenomenon of focal dominance, and that the frequency of susceptible hosts in the initial outbreak focus had a stronger influence when the proportion of propagules that remained local during dispersal was higher. Overall, our results suggest that targeting pathogen reproduction in the initial outbreak focus will have a disproportionately large impact on subsequent epidemic spread.


title: [Standard procedures in dialysis during the Covid-19 epidemic].

text: Scope: The aim of this document has been to define standard procedures for dealing with dialysis patients once the first cases of novel Coronavirus 2019-nCoV (Covid-19) were confirmed among the Italian population. Applicability: These procedures, that refer exclusively to the hospital's dialysis rooms, are currently implemented at the ASST Santi Paolo e Carlo in Milan and two smaller centers in the Milan area. Description: We describe the preemptive measures adopted by the staff at our dialysis unit since 24/02/2020, in order to slow down the transmission of Covid-19. They have allowed us to adopt a uniform approach towards all patients, streamlining the way we identify and deal with suspected, likely and confirmed Coronavirus infections. To start with, all patients coming to the hospital for their dialytic session have been treated as potentially infectious and everybody has been following closely the standard protocols regarding personal protective equipment (PPE).


Crossing the language limitations.


title: Why a burden of disease study?

text: From the time I was appointed as Director of the European Centre for Disease Prevention and Control (ECDC) in 2005, I and the ECDC Governing and Advisory Bodies faced the task of tackling the 46 diseases under mandatory notification in the European Union (EU), as well as severe acute respiratory syndrome (SARS), avian influenza and West Nile virus.


title: Glycoprotein D-independent infectivity of pseudorabies virus results in an alteration of in vivo host range and correlates with mutations in glycoproteins B and H.

text: Infection of cells by herpesviruses is initiated by the interaction of viral envelope glycoproteins with cellular receptors. In the alphaherpesvirus pseudorabies virus (PrV), the causative agent of Aujeszky's disease in pigs, the essential glycoprotein D (gD) mediates secondary attachment of virions to target cells by binding to newly identified cellular receptors (R. J. Geraghty, C. Krummenacher, G. H. Cohen, R. J. Eisenberg, and P. G. Spear, Science 280:1618-1620, 1998). However, in the presence of compensatory mutations, infection can also occur in the absence of gD, as evidenced by the isolation in cell culture of an infectious gD-negative PrV mutant (PrV-gD(-) Pass) (J. Schmidt, B. G. Klupp, A. Karger, and T. C. Mettenleiter, J. Virol. 71:17-24, 1997). PrV-gD(-) Pass is replication competent with an only moderate reduction in specific infectivity but appears to bind to receptors different from those recognized by wild-type PrV (A. Karger, J. Schmidt, and T. C. Mettenleiter, J. Virol. 72:7341-7348, 1998). To analyze whether this alteration in receptor usage in vitro influences infection in vivo, the model host mouse and the natural host pig were intranasally infected with PrV-gD(-) Pass and were compared to animals infected by wild-type PrV. For mice, a comparable progress of disease was observed, and all animals infected with mutant virus died, although they exhibited a slight delay in the onset of symptoms and, correspondingly, a longer time to death. In contrast, whereas wild-type PrV-infected pigs showed clinical signs and histological and histopathological findings typical of PrV infection, no signs of disease were observed after infection with PrV-gD(-) Pass. Moreover, in these animals, virus-infected cells were not detectable by immunohistochemical staining of different organ samples and no virus could be isolated from nasal swabs. Mutations in glycoproteins B and H were found to correlate with, and probably contribute to, gD-independent infectivity. In conclusion, although PrV-gD(-) Pass is virulent in mice, it is apparently unable to infect the natural host, the pig. This altered host range in vivo correlates with a difference of receptor usage in vitro and demonstrates for the first time the importance of gD receptors in alphaherpesvirus infection of an animal host.


title: The esterase and PHD domains in CR1-like non-LTR retrotransposons.

text: Most active non-LTR (long terminal repeat) retrotransposons carry two open reading frames (ORFs) encoding ORF1p and ORF2p proteins. The ORF2p proteins are relatively well studied and are known to contain endonuclease/reverse transcriptase domains. At the same time, the biological function of ORF1p proteins remains poorly understood, except in that they nonspecifically bind single-stranded mRNA/DNA molecules. CR1-like elements form the most widely distributed clade/superfamily of non-LTR retrotransposons. We found that ORF1p proteins encoded by diverse CR1-like elements contain conserved esterase domain (ES) or plant homeodomain (PHD). This indicates that CR1-like ORF1p proteins are either lipolytic enzymes or are involved in protein-protein interactions related to chromatin remodeling. Sequence conservation of ES suggests that interaction with cellular membranes is an important phase in life circles of CR1-like elements. Presumably such interaction helps in penetrating host cells. As a consequence, the presence of multiple young CR1 families characterized by approximately 10% intrafamily and 40% interfamily identities may be explained by a relatively frequent horizontal transfer of these CR1-like elements. Unexpectedly, ES links together non-LTR retrotransposons and single-stranded RNA viruses like influenza C and coronaviruses, which are known to depend on their own ES.


The value of a human rights perspective in health and foreign policy.


title: [Implementation of Remote Clinics in urology practice during the COVID-19 era: What have we learned?]

text: The COVID-19 pandemic has requiredd rastic measures for an attempt in controlling its spread. Health resources and facilities are being destined for the treatment of critically ill infected patients. During the past weeks, we, as urologists have faced increasingly difficult changes in practice, as out patient activity and elective surgeries must be postponed in order to save resources and limit the mobilization of patients and faculty. During this conflictive situation, telehealth medicine can provide adequate support using technological tools and trying to simulate face-to-face consults with the use of video or telephone calls. However, many out patient clinics and facilities are not ready yet for telehealth as their experience in this area is low. The benefits for telemedicine in urology are continuing urologic outpatient follow-up, providing recommendations and prescriptions, and the triage of patients who will need urgent procedures. Urology residency training has suffered an abrupt disruption nowadays as outpatient, surgical and academic meetings are cancelled. In this scenario, virtual strategies and "smart learning" activities are being used to continue education. We provide a review of the latest published literature regarding the use of telehealth medicine or telemedicine for the modern urology practice, along side our recommendations and conclusions.


title: Hospital disinfection: efficacy and safety issues.

text: PURPOSE OF REVIEW To review recent publications relevant to hospital disinfection (and cleaning) including the reprocessing of medical instruments. RECENT FINDINGS The key question as to whether the use of disinfectants on environmental surfaces rather than cleaning with detergents only reduces nosocomial infection rates still awaits conclusive studies. New disinfectants, mainly peroxygen compounds, show good sporicidal properties and will probably replace more problematical substances such as chlorine-releasing agents. The safe reprocessing of medical devices requires a well-coordinated approach, starting with proper cleaning. New methods and substances show promising activity for preventing the transmission of prions. Different aspects of virus inactivation have been studied, and the transmissibility, e.g. of norovirus, shows the need for sound data on how different disinfectant classes perform. Biofilms or other forms of surface-adherent organisms pose an extraordinary challenge to decontamination. Although resistance to biocides is generally not judged to be as critical as antibiotic resistance, scientific data support the need for proper use, i.e. the avoidance of widespread application, especially in low concentrations and in consumer products. SUMMARY Chemical disinfection of heat-sensitive instruments and targeted disinfection of environmental surfaces are established components of hospital infection control. To avoid danger to staff, patients and the environment, prudent use as well as established safety precautions are required. New technologies and products should be evaluated with sound methods. As emerging resistant pathogens will challenge healthcare facilities in the future even more than at present, there is a need for well-designed studies addressing the role of disinfection in hospital infection control.


title: Targets of anticytokine therapy and the risk of infections in humans and mice.

text: PURPOSE OF REVIEW To examine the type and risk of infections in humans and mice deficient in proinflammatory cytokines. Naturally occurring or manipulated genetic defects of tumor necrosis factor, interleukins-1, -6, -12, and -15, and interferon-gamma are examined for their increased susceptibility to, or protection from, infection. RECENT FINDINGS Interleukin-12p40 and interferon-gamma-blockers may lead to increased incidence of infections with intracellular bacteria, parasites, and fungi. In addition, we may see viral infections with interferon-gamma-blockers. Increased risk of infections is unlikely with either interleukin-1- or interleukin-15-blockers. Interleukin-6-blockers may lead to increased risk of infection with extracellular bacteria, viruses, parasites and fungi. SUMMARY In tumor necrosis factor knockout mice, increased susceptibility to pathogens are reported that are normally controlled by granuloma formation. In patients treated with tumor necrosis factor-blockers, a two-fold increase of granulomatous infections, predominantly reactivation of latent tuberculosis, is found. The infections detected in tumor necrosis factor knockout mice were accurate for predicting the infections observed when using tumor necrosis factor-blockers. If a similar correlation exists for other cytokines, the use of interferon-gamma and interleukin-12p40 blockers, and possibly interleukin-6 blockers, will lead to an increased risk for severe infections. Care should be taken when new cytokine blockers/antagonists are introduced.


title: Pathogenic characterization of a cervical lymph node derived from a patient with Kawasaki disease.

text: Kawasaki disease (KD) is the most common cause of multisystem vasculitis in childhood. Although cervical lymphadenitis is one of the major symptoms in KD, lymph node biopsy is rarely performed, because KD is usually diagnosed by clinical symptoms. A cervical lymph node biopsy was taken from a girl aged 1 year and 8 months who had suspected lymphoma, but she was diagnosed with KD after the biopsy. The cervical lymph node specimen was analyzed with multivirus real-time PCR that can detect >160 viruses, and unbiased direct sequencing with a next-generation DNA sequencer to detect potential pathogens in the lymph node. Histologically, focal necrosis with inflammatory cell infiltration, including neutrophils and macrophages, was observed in the marginal zone of the cervical lymph node, which was compatible with the acute phase of KD. Multivirus real-time PCR detected a low copy number of torque teno virus in the sample. Comprehensive direct sequencing of the cervical lymph node biopsy sample sequenced more than 8 million and 3 million reads from DNA and RNA samples, respectively. Bacterial genomes were detected in 0.03% and 1.79% of all reads in DNA and RNA samples, respectively. Although many reads corresponded to genomes of bacterial environmental microorganisms, Streptococcus spp. genome was detected in both DNA (77 reads) and RNA (2,925 reads) samples. Further studies are required to reveal any association of microbial or viral infection with the pathogenesis of KD.


title: Efficacy of severe acute respiratory syndrome vaccine based on a nonhuman primate adenovirus in the presence of immunity against human adenovirus.

text: Replication-deficient human adenovirus type 5 (AdH5) vectors can induce strong transgene product-specific cellular and humoral responses. However, many adult humans have neutralizing antibodies (NAbs) against AdH5 as a result of natural infection with this virus. Therefore, a chimpanzee adenovirus C7 (AdC7) vector was developed to circumvent interference by preexisting immunity to AdH5. This study evaluated the impact of preexisting immunity to human adenovirus on the efficacy of adenovirus-based vaccines against the coronavirus that causes severe acute respiratory syndrome (SARS-CoV). Efficacy was assessed after intramuscular injection of the vector into mice and was measured as the frequency of SARS-CoV-specific T cells and NAbs against SARS-CoV. Immunogenicity of the AdH5-based vaccine was significantly attenuated or completely abolished when the preexisting anti-AdH5 NAb titer was higher than 40. Because 27% of human serum samples from the United States tested so far have an anti-AdH5 NAb titer higher than 40, our results suggested that a significant percentage of humans with preexisting anti-AdH5 immunity would not be candidates for vaccination with an AdH5-based genetic vaccine. In contrast, preexisting anti-AdH5 NAbs have a minimal effect on the potency of the AdC7-based genetic vaccine. Taken together, our studies warrant the further development of AdC7 as a vaccine carrier for human trials.


title: The pathogenic role of virus-specific antibody-secreting cells in the central nervous system of rats with different susceptibility to coronavirus-induced demyelinating encephalitis.

text: The humoral immune response in the central nervous system (CNS) of susceptible Lewis (LE) rats and resistant Brown Norway (BN) rats was analysed after intracerebral infection with the murine coronavirus JHM (MHV4). The subclinical course of the infection in BN rats was characterized by an early rise of neutralizing antibodies in the cerebrospinal fluid (CSF) 7 days post-infection. At this time in LE rats, neutralizing antibodies were not detectable in the CSF and the animals developed neurological signs of infection. Subsequently, LE rats recovered from disease. This process was accompanied by increasing titres of virus-neutralizing antibodies. Within the CNS parenchyma of both rat strains, equivalent numbers of IgM-secreting cells were detected. However, in BN rats, virus-specific IgG secreting cells appeared earlier and in higher numbers. Moreover, based on the size of zones of antibody secreted by single cells in the Spot-ELISA assay, it appeared that cells from BN rats secreted IgG antibody of higher affinity. These data suggest that early maturation of antiviral antibody responses in the resistant BN rat probably restricts the spread of viral infection to small foci within the CNS, resulting in a subclinical level of primary demyelination. In contrast, the absence of neutralizing antibodies in the susceptible LE rats favours spread of the virus throughout the CNS, resulting finally in severe neurological disease.


Canada reports more than 300 suspected cases of SARS.


title: Two related strains of feline infectious peritonitis virus isolated from immunocompromised cats infected with a feline enteric coronavirus.

text: Two groups of cats were experimentally infected orally with the cat-passaged RM strain of feline enteric coronavirus (FECV-RM). One group of cats (n = 19) had been chronically infected with feline immunodeficiency virus (FIV) for over 6 years, while a second control group (n = 20) consisted of FIV-naive siblings. Fecal virus shedding of FECV occurred in both groups starting on day 3 postinfection, nearly ceased by 4 weeks in FIV-uninfected cats, but remained at high levels in FIV-infected animals. FIV-infected cats shed virus for a longer period of time and at levels 10 to 100 times greater than those for FIV-uninfected cats. The coronavirus antibody response of the FIV-infected cats was delayed and of reduced titer compared with that of the FIV-uninfected animals. Cats in both groups remained asymptomatic for the first two months following FECV-RM infection; however, 8 to 10 weeks postinfection two cats in the FIV-infected group developed feline infectious peritonitis (FIP). The FIP viruses (designated FIPV-UCD9 and -UCD10) isolated from these two cats had almost complete genetic homology to each other and to the infecting FECV-RM. However, unlike FECV-RM, they readily induced FIP when inoculated intraperitoneally into specific-pathogen-free cats. This study confirms that FIPVs are frequently and rapidly arising mutants of FECV. Immunosuppression caused by chronic FIV infection may have enhanced the creation and selection of FIPV mutants by increasing the rate of FECV replication in the bowel and inhibiting the host's ability to combat the mutant viruses once they occurred.


[Brug af hydroxychloroquin til COVID-19].


title: CD4+ and CD8+ T cells are not major effectors of mouse hepatitis virus A59-induced demyelinating disease.

text: We examined murine hepatitis virus strain A59 (MHV-A59)-induced demyelinating disease in C57BL/6 mice which had previously been thymectomized at 25 days of age. Demyelination was observed in 51-96% of spinal cord quadrants examined 30 or 60 days post infection (dpi), indicating that neither an intact thymus nor thymic infection is a prerequisite to demyelination. Depletion of CD4+ or CD8+ T cells at 5, 7 or 10 dpi did not influence the extent of demyelination indicating that neither T cell subset is a major effector of demyelination. However, these findings do not exclude the possibility that T cells are involved in initiating demyelinating disease very early in infection.


title: [Elements of state coercion in health care: selected issues].

text: Polish system of public law provides for the possibility of forced subjection of a person to a medical procedure, both in case of infectious and mental illnesses. The authors discuss the cases of infectious illnesses, subject to compulsory treatment in outpatient medical care and to compulsory hospitalization, as well as the procedure protocol in case of suspicion or corroboration of an infectious illness case. Another reason for a physician's authorization to initiate a compulsory medical procedure without the patient's consent is suspicion of a mental illness; whenever the patients' behavior can endanger them, other persons or their immediate surroundings. Introduced is also a detailed description of actual status in order to illustrate an appropriate application of coercion means.


PATHOGENESIS of fever.


Preventing the introduction of Ebola virus into the Eastern Mediterranean Region: enhanced preparedness is the key.


title: The p23 protein of citrus tristeza virus controls asymmetrical RNA accumulation.

text: Citrus tristeza virus (CTV), a member of the Closteroviridae, has a 19.3-kb positive-stranded RNA genome that is organized into 12 open reading frames (ORFs) with the 10 3' genes expressed via a nested set of nine or ten 3'-coterminal subgenomic mRNAs (sgRNAs). Relatively large amounts of negative-stranded RNAs complementary to both genomic and sgRNAs accumulate in infected cells. As is characteristic of RNA viruses, wild-type CTV produced more positive than negative strands, with the plus-to-minus ratios of genomic and sgRNAs estimated at 10 to 20:1 and 40 to 50:1, respectively. However, a mutant with all of the 3' genes deleted replicated efficiently, but produced plus to minus strands at a markedly decreased ratio of 1 to 2:1. Deletion analysis of 3'-end genes revealed that the p23 ORF was involved in asymmetric RNA accumulation. A mutation which caused a frameshift after the fifth codon resulted in nearly symmetrical RNA accumulation, suggesting that the p23 protein, not a cis-acting element within the p23 ORF, controls asymmetric accumulation of CTV RNAs. Further in-frame deletion mutations in the p23 ORF suggested that amino acid residues 46 to 180, which contained RNA-binding and zinc finger domains, were indispensable for asymmetrical RNA accumulation, while the N-terminal 5 to 45 and C-terminal 181 to 209 amino acid residues were not absolutely required. Mutation of conserved cysteine residues to alanines in the zinc finger domain resulted in loss of activity of the p23 protein, suggesting involvement of the zinc finger in asymmetric RNA accumulation. The absence of p23 gene function was manifested by substantial increases in accumulation of negative-stranded RNAs and only modest decreases in positive-stranded RNAs. Moreover, the substantial decrease in the accumulation of negative-stranded coat protein (CP) sgRNA in the presence of the functional p23 gene resulted in a 12- to 15-fold increase in the expression of the CP gene. Apparently the excess negative-stranded sgRNA reduces the availability of the corresponding positive-stranded sgRNA as a messenger. Thus, the p23 protein controls asymmetric accumulation of CTV RNAs by downregulating negative-stranded RNA accumulation and indirectly increases expression of 3' genes.


title: Genomic imprinting in disruptive spermatogenesis.

text: The possibility of imprinting disease transmission by assisted reproductive technologies has been raised after births of children with Angelman's and Beckwith-Wiedemann's syndromes. To investigate whether imprinting defects were associated with disturbed spermatogenesis, we studied two oppositely imprinted genes in spermatozoan DNA from normozoospermic and oligozoospermic patients. In the mesodermal specific transcript gene (MEST), bisulphite genomic sequencing showed that maternal imprinting was correctly erased in all 123 patients. However, methylation of the H19 gene did not change in any of 27 normozoospermic individuals (0%, 95% CI 0-13%), compared with methylation changes in eight moderate (17%, 8-31%, p=0.026) and 15 severe (30%, 18-45%, p=0.002) oligozoospermic patients. Our data suggest an association between abnormal genomic imprinting and hypospermatogenesis, and that spermatozoa from oligozoospermic patients carry a raised risk of transmitting imprinting errors.


title: Formalin-treated UV-inactivated SARS coronavirus vaccine retains its immunogenicity and promotes Th2-type immune responses.

text: The demand for rapid and simple development of a vaccine against a newly emerging infectious disease is increasing worldwide. We previously revealed that UV-inactivated severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) virions (UV-V) elicited high levels of humoral immunity and a weak Th0 response in mice immunized subcutaneously. To ensure the safety of such a whole inactivated SARS-CoV vaccine, we additionally treated the UV-V vaccine with formalin, resulting in the UV-F-V vaccine. Analysis of the immunogenicity of the UV-F-V+alum vaccine in mice revealed that it generated comparable neutralizing serum anti-SARS-CoV IgG antibody levels as the UV-V+alum vaccine. Moreover, both vaccines induced similar frequencies of anti-SARS-CoV IgG antibody-producing cells in bone marrow. Interestingly, the UV-F-V vaccine induced fewer IgG(2a) subtype antibodies and higher interleukin-4 production in vaccinated mice than did UV-V. Thus, UV-F-V imposes a Th2-type bias on the immune response, unlike UV-V. We propose here that doubly-inactivated SARS-CoV virions by UV and formalin constitute a safe vaccine that may effectively induce neutralizing antibodies in humans.


title: Television and video viewing time among children aged 2 years - Oregon, 2006-2007.

text: Excessive exposure of children to television and videos (viewing time) is associated with impaired childhood development and childhood obesity. In 2001, the American Academy of Pediatrics (AAP) recommended that children watch no more than 1 to 2 hours of "quality programming" per day, and that televisions be removed from children's bedrooms. To determine the risk for excessive viewing time among children aged 2 years, CDC and the Oregon Public Health Division analyzed 2006 and 2007 data from the Oregon Pregnancy Risk Assessment Monitoring Survey follow-back survey (Oregon PRAMS-2), which was used to re-interview mothers who had participated in PRAMS. This report summarizes the results of that analysis, which indicated that, on a typical day, 19.6% of children aged 2 years spent >or=2 hours watching television or videos. A total of 18.2% of children had a television in their bedroom; these children were more likely to have >or=2 hours viewing time compared with children without a television in the bedroom (34.1% versus 16.3%). In multivariable analysis, >or=2 hours of viewing time was positively associated with the presence of a television in the child's bedroom, non-Hispanic black maternal race/ethnicity, fewer than four outings with the child during the preceding week, and was negatively associated with obtaining child care in a child care center. In Oregon, these findings support the AAP recommendations that health professionals, parents, and caregivers recognize the extent of children's media consumption, and that televisions be removed from children's bedrooms. Other states should consider conducting similar surveys.


title: Identification of mixed infections with different genotypes of avian bornaviruses in psittacine birds with proventricular dilatation disease.

text: Proventricular dilatation disease (PDD) is a fatal, progressive neurological disorder of psittacine birds, which is caused by a single-stranded RNA virus, the avian bornavirus (ABV). The disease pattern includes lymphoplasmacytic inflammation of the central, peripheral and autonomic nervous system. Seven avian bornavirus genotypes have been identified during the last years. So far only monoinfections with a single genotype of ABV have been attributed to PDD cases. However, after a recent survey discovered a case of a double infection with two different ABV genotypes, this seemed to indicate the need for a more systematic search for mixed infections. Brain specimens from 21 psittacine birds affected with PDD were examined. Aim of the investigation was to generate partial ABV sequences of a part of the matrix protein (M) gene and to evaluate whether sequences of more than one ABV genotype were present. RNA was extracted, and subjected to reverse transcriptase PCR with primer pairs generating a partial sequence of the matrix protein (M) gene, followed by a cloning procedure. Ten clones per case were sequenced in order to elucidate whether sequences characteristic for one or more than one genotype were present. In 19 of 21 cases clear M gene sequences could be generated; in two cases nucleic acid amplification failed. Seven birds were infected with ABV 2 and nine with ABV 4, representing the predominant genotypes in Europe. Two cases showed a mixed infection with ABV 2 and ABV 4, and one case a mixed infection with ABV 2 and ABV 6. These results suggest that the molecular cloning method is a useful tool for distinguishing between single and multiple infection events by different ABV genotypes.


title: Entry and release of transmissible gastroenteritis coronavirus are restricted to apical surfaces of polarized epithelial cells.

text: The transmissible gastroenteritis coronavirus (TGEV) infects the epithelial cells of the intestinal tract of pigs, resulting in a high mortality rate in piglets. This study shows the interaction of TGEV with a porcine epithelial cell line. To determine the site of viral entry, LLC-PK1 cells were grown on permeable filter supports and infected with TGEV from the apical or basolateral side. Initially after plating, the virus was found to enter the cells from both sides. During further development of cell polarity, however, the entry became restricted to the apical membrane. Viral entry could be blocked by a monoclonal antibody to the viral receptor aminopeptidase N. Confocal laser scanning microscopy showed that this receptor protein was present at both the apical and basolateral plasma membrane domains just after plating of the cells but that it became restricted to the apical plasma membrane during culture. To establish the site of viral release, the viral content of the apical and basolateral media of apically infected LLC-PK1 cells was measured by determining the amount of radioactively labelled viral proteins and infectious viral particles. We found that TGEV was preferentially released from the apical plasma membrane. This conclusion was confirmed by electron microscopy, which demonstrated that newly synthesized viral particles attached to the apical membrane. The results support the idea that the rapid lateral spread of TGEV infection over the intestinal epithelia occurs by the preferential release of virus from infected epithelial cells into the gut lumen followed by efficient infection of nearby cells through the apical domain.


title: [Etiology of infectious diseases in pregnant women with symptoms of fever and lung syndrome].

text: 31 samples of blood serum of pregnant women with acute respiratory virus infection and pneumonia symptoms who had been administered into hospital were analyzed. Antibacterial and antifungal treatment was ineffective. Autumn-winter-spring seasonality was detected. In connection with above-mentioned, an attempt was made to find out the etiology of a disease using viroloy methods. Primers of hantaviruses, Crimea-and-Congo hemorrhage fever, coronaviruses, SARS, Leptospirosis and virus hepatitis. Genome of B hepatitis virus was discovered in 3 samples, C--in one sample, G--in four samples. The test for other primers was negative. It was shown that in case of serious clinical course only abortion can save mother's life. It is necessary to find out aetiology of the disease, primers and other pathogenic organisms of virus infections.


title: Plaque assay and improved yield of human coronaviruses in a human rhabdomyosarcoma cell line.

text: Propagation and plaque assay of human coronavirus prototypes were studied in two human cell lines: a diploid fetal tonsil (FT) and a heteroploid rhabdomyosarcoma (RD) cell lines. Plaques, observed within 2 to 3 days on FT cell monolayers with both 229E and OC43 viruses, appeared as colorless areas after staining with neutral red or crystal violet, whereas neutral red staining was required for visualization of plaques on RD cells. The plating efficiencies were approximately equal between the two cell lines, but virus assay by plaque formation was 15- to 30-fold more efficient than tube dilution assay with 50% endpoints. The discrepancy between 50% endpoint and plaque-forming unit values was striking and appeared to result from the fact that killing of cells (particularly RD cells) by coronaviruses was not accompanied by visible changes in the cells but killing was detected by the failure of infected cells to stain with a vital dye. The latent phase in one-step growth curves was 5 to 6 h for both viruses in either cell line, but the maximum yield of intracellular virus was reached in 18 to 20 h for FT cells and 24 to 28 h for RD cells. Virus release also differed between the two cell lines: in FT cells, the maximum yield of extracellular virus was reached 2 to 3 h later than that of intracellular virus, whereas in RD cells, the difference was 5 h for 229E virus and 10 h for OC43 virus. Although both cell lines appear equally useful for plaque assay, RD cells would be preferred for mass virus propagation because yields (5 X 10(8) plaque-forming units per ml) were 10-fold higher than in FT cells, a finding true for both virus prototypes.


title: Severe influenza among children and young adults with neurologic and neurodevelopmental conditions - Ohio, 2011.

text: Children with neurologic and neurodevelopmental conditions are at increased risk for severe outcomes from influenza, including death. In April 2011, the Ohio Department of Health and CDC investigated an influenza outbreak that began in February 2011 in a residential facility for 130 children and young adults with neurologic and neurodevelopmental conditions. This report summarizes the characteristics and clinical courses of 13 severely ill residents with suspected or confirmed influenza; 10 were hospitalized, and seven died. Diagnosis is challenging in this population, and clinicians should consider influenza in patients with neurologic and neurodevelopmental conditions who have respiratory illness or a decline in baseline medical status when influenza is circulating in the community. Prompt testing, early and aggressive antiviral treatment, and antiviral chemoprophylaxis are important for these patients. When influenza is suspected, antiviral treatment should be given as soon as possible after symptom onset, ideally within 48 hours. Treatment should not wait for laboratory confirmation of influenza. During outbreaks, antiviral chemoprophylaxis should be provided to all residents of institutional facilities (e.g., nursing homes and long-term- care facilities), regardless of vaccination status. Residential facilities for patients with neurologic and neurodevelopmental conditions are encouraged to vaccinate all eligible residents and staff members against influenza.


title: Detection of classical swine fever virus in boar semen by reverse transcription-polymerase chain reaction.

text: A seminested reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of classical swine fever virus (CSFV) in semen. Five boars were inoculated intranasally with CSFV isolate propagated in PK15 cells. Two boars inoculated with the supernatant of noninfected PK15 cells were kept as controls. Semen and serum samples were collected twice weekly for 63 days postinoculation (dpi). Samples were tested for the presence of antibodies to CSFV by an enzyme-linked immunosorbent assay and for the presence of CSFV nucleic acid by seminested RT-PCR. Antibodies to CSFV could be detected as early as 7 dpi in 1 boar, and all 5 infected boars were found positive by 14 dpi. CSFV from boar semen was infrequently identified by virus isolation compared with seminested RT-PCR. CSFV nucleic acid was detected in semen by seminested RT-PCR as early as 7 dpi in 3 infected boars and persistently thereafter in all 5 infected boars until 63 dpi. When separated fractions of CSFV-contaminated semen were analyzed by the seminested RT-PCR, the CSFV nucleic acid was detected mainly in seminal fluid and occasionally in nonsperm cells. CSFV antigen was also detected in nonsperm cells from semen smear by immunohistochemistry. Thus, infection via semen, specially through CSFV-infected seminal fluid, seems to be a major route of transmission of CSFV.


title: Passive transfer of virus-specific antibodies confers protection against reproductive failure induced by a virulent strain of porcine reproductive and respiratory syndrome virus and establishes sterilizing immunity.

text: Immune mechanisms mediating protective immunity against porcine reproductive and respiratory syndrome virus (PRRSV) are not well understood. The PRRSV-specific humoral immune response has been dismissed as being ineffective and perhaps deleterious for the host. The function of PRRSV antibodies in protective immunity against infection with a highly abortifacient strain of this virus was examined by passive transfer experiments in pregnant swine. All of a group of pregnant gilts (n = 6) that received PRRSV immunoglobulin (Ig) from PRRSV-convalescent, hyperimmune animals were fully protected from reproductive failure as judged by 95% viability of offspring at weaning (15 days of age). On the other hand, the totality of animals in a matched control group (n = 6) receiving anti-pseudorabies virus (PRV) Ig exhibited marked reproductive failure with 4% survival at weaning. Besides protecting the pregnant females from clinical reproductive disease, the passive transfer of PRRSV Ig prevented the challenge virus from infecting the dams and precluded its vertical transmission, as evidenced by the complete absence of infectious PRRSV from the tissues of the dams and lack of infection in their offspring. In summary, these results indicate that PRRSV-Igs are capable of conferring protective immunity against PRRSV and furthermore that these Igs can provide sterilizing immunity in vivo.


title: Minimal region sufficient for genome dimerization in the human immunodeficiency virus type 1 virion and its potential roles in the early stages of viral replication.

text: It has been suggested that the dimer initiation site/dimer linkage sequence (DIS/DLS) region of the human immunodeficiency virus type 1 (HIV-1) RNA genome plays an important role at various stages of the viral life cycle. Recently we found that the duplication of the DIS/DLS region on viral RNA caused the production of partially monomeric RNAs in virions, indicating that this region indeed mediates RNA-RNA interaction. In this report, we followed up on this finding to identify the necessary and sufficient region for RNA dimerization in the virion of HIV-1. The region thus identified was 144 bases in length, extending from the junction of R/U5 and U5/L stem-loops to the end of SL4. The trans-acting responsive element, polyadenylation signal, primer binding site, upper stem-loop of U5/L, and SL2 were not needed for the function of this region. The insertion of this region into the ectopic location of the viral genome did not affect the level of virion production by transfection. However, the resultant virions contained monomerized genomes and showed drastic reductions in infectivity. A reduction was observed especially in the reverse transcription process. An attempt to generate a replication-competent virus with monomerized genome was performed by the long-term culture of mutant virus-infected cells. All recovered viruses were wild-type revertants, indicating a fatal defect of the mutation. These results suggest that genome dimerization or DIS/DLS itself also plays an important role in the early stages of virus infection.


title: Development of an antigen spot test for detection of coronavirus in bovine fecal samples.

text: We have developed a rapid and sensitive microimmunodot blot assay, the antigen spot test (AST), for the detection of bovine coronavirus (BCV) antigen from neonatal calf fecal samples. The AST procedure can be completed in 3.5 h, whereas the previously reported immunodot blot assays require 10 to 12 h. Ninety-six samples can be tested per membrane, and 10 membranes (960 samples) may be processed by a single technologist in 1 working day. The effects of detergents, oxidizing chemicals, chaotropic agents, and enzyme substrates in improving the sensitivity and signal-to-noise ratio of the AST were studied. Finally, the sensitivity and specificity of AST for the detection of BCV antigen were compared to those of a sandwich enzyme-linked immunosorbant assay (ELISA) and a hemagglutination assay (HA). Of 347 field samples tested by all three methods, 94.2% were positive by AST, 91.4% were positive by ELISA, and 86.7% were positive by HA. The sensitivity of the AST was determined to be 100% compared to the results of the ELISA reference method. The specificity of the AST was 67%, which reflects a lower limit of detection of 10(4) viral particles per ml in a 10% fecal suspension.


title: Induction of prothrombinase fgl2 by the nucleocapsid protein of virulent mouse hepatitis virus is dependent on host hepatic nuclear factor-4 alpha.

text: Fibrinogen-like protein 2/fibroleukin (Fgl2) plays a pivotal role in the pathogenesis of both experimental and human fulminant hepatic failure. We have reported recently that the nucleocapsid (N) protein from strains of murine hepatitis virus (MHV-3, MHV-A59), which cause massive hepatocellular necrosis but not from strains (MHV-JHM, MHV-2) which do not produce serious liver disease, induces transcription of fgl2. The purpose of the present study was to characterize both viral and host factor(s) necessary for viral induced transcription of fgl2. Mutation of residues Gly-12, Pro-38, Asn-40, Gln-41, and Asn-42 within domain 1 of the N protein of MHV-A59 to their corresponding residues found in MHV-2 abrogated fgl2 transcription, whereas mutation of other N protein domains, including a protein expressed from an internal reading frame (I protein), did not affect fgl2 gene transcription. We then examined the -372 to -306 sequence within the 1.3-kb fgl2 promoter region upstream from the transcription start site that was previously identified as necessary for N protein-induced gene transcription. We demonstrated that the -331/-325 HNF4 cis-element and its cognate transcription factor, HNF4alpha, are necessary for virus-induced fgl2 gene transcription. In uninfected macrophages and macrophages infected with MHV-2, an unidentified protein occupies the HNF4 cis-element. Following stimulation with MHV-A59, it was shown by electrophoretic mobility shift assay that HNF4alpha binds the HNF4 cis-element in the fgl2 promoter. We further report the unprecedented presence of HNF4alpha in peritoneal macrophages. Collectively, the results of this study define both viral and host factors necessary for induction of fgl2 prothrombinase gene transcription in MHV infection and may provide an explanation for the hepatotrophic nature of MHV-induced fulminant hepatic failure.


title: [Exploration and practice of online teaching system of medical immunology based on small private online course (SPOC) model during coronavirus disease 2019 (COVID-19) pandemic].

text: During the pandemic of coronavirus disease 2019 (COVID-19), the Chinese Ministry of Education put forward the requirement of turning offline education into online teaching to universities. Our department positively responded and rapidly mobilized to establish an online teaching system of medical immunology based on the small private online course (SPOC) model, which has a main body of the SPOC model with "video course as the main part, supplemented by online Q&A". The system also has feedback improvement sections of online discussion and chapter tests and evaluates the quality and effect of teaching with students' group display, course examination, and questionnaires. At the same time, the COVID-19 hotspot is also organically combined with immunological knowledge throughout the online teaching. In this process, our department continuously summarizes experiences and discovers problems. Herein, we generalize and sort out the exploration and practice of constructing the online teaching system, to carry out offline teaching after the pandemic and to continue this teaching model in the future, for reference and guidance.


title: Laparoscopic total extraperitoneal repair under spinal anesthesia versus general anesthesia: a randomized prospective study.

text: BACKGROUND Laparoscopic total extraperitoneal (TEP) inguinal hernia repair is a well-known approach to inguinal hernia repair that is usually performed under general anesthesia (GA). To date, no reports compare the efficacy of spinal anesthesia (SA) with that of GA for laparoscopic hernia repairs. The purpose of this study was to compare the surgical outcome of TEP inguinal hernia repair performed when the patient was treated under SA with that performed under GA. MATERIALS AND METHODS Between July 2015 and July 2016, 50 patients were prospectively randomized to either the GA TEP group (Group I) or the SA TEP group (Group II). Propofol, fentanyl, rocuronium, sevoflurane, and tracheal intubation were used for GA. Hyperbaric bupivacaine (15 mg) and fentanyl (10 µg) were used for SA to achieve a sensorial level of T3. Intraoperative events related to SA, operative and anesthesia times, postoperative complications, and pain scores were recorded. Each patient was asked to evaluate the anesthetic technique by using a direct questionnaire filled in 3 months after the operation. RESULTS All the procedures were completed by the allocated method of anesthesia as there were no conversions from SA to GA. Pain was significantly less for 1 h (P<0.0001) and 4 h (P=0.002) after the procedure for the SA and GA groups, respectively. There was no difference between the two groups regarding complications, hospital stay, recovery, or surgery time. Generally, patients were more satisfied with SA than GA (P<0.020). CONCLUSION TEP inguinal hernia repair can be safely performed under SA, and SA was associated with less postoperative pain, better recovery, and better patient satisfaction than GA.


title: Ring structure of the Escherichia coli DNA-binding protein RdgC associated with recombination and replication fork repair.

text: The DNA-binding protein, RdgC, is associated with recombination and replication fork repair in Escherichia coli and with the virulence-associated, pilin antigenic variation mediated by RecA and other recombination proteins in Neisseria species. We solved the structure of the E. coli protein and refined it to 2.4A. RdgC crystallizes as a dimer with a head-to-head, tail-to-tail organization forming a ring with a 30 A diameter hole at the center. The protein fold is unique and reminiscent of a horseshoe with twin gates closing the open end. The central hole is lined with positively charged residues and provides a highly plausible DNA binding channel consistent with the nonspecific mode of binding detected in vitro and with the ability of RdgC to modulate RecA function in vivo.


title: Assessing the impact of travel restrictions on international spread of the 2014 West African Ebola epidemic.

text: The quick spread of an Ebola outbreak in West Africa has led a number of countries and airline companies to issue travel bans to the affected areas. Considering data up to 31 Aug 2014, we assess the impact of the resulting traffic reductions with detailed numerical simulations of the international spread of the epidemic. Traffic reductions are shown to delay by only a few weeks the risk that the outbreak extends to new countries.


title: Autophagy is involved in influenza A virus replication.

text: Autophagy is involved in the replication of viruses, especially those that perform RNA assembly on the surface of cytoplasmic membrane in host cells. However, little is known about the regulatory role of autophagy in influenza A virus replication. Using fluorescence and electron microscopy, we observed that autophagosomes can be induced and identified upon influenza A virus infection. The virus increased the amount of the autophagosome marker protein microtubule-associated protein light chain 3-II (LC3-II) and enhanced autophagic flux. When autophagy was pharmacologically inhibited by either 3-methylademine or wortmannin, the titers of influenza A virus were remarkably decreased. Viral reduction via autophagy inhibition was further confirmed by RNA interference, through which two different proteins required for autophagy were depleted. Noticeably, the compounds utilized had no marked effect on virus entry or cell viability, either of which might limit viral replication. Furthermore, alteration of cellular autophagy via pharmacological reagents or RNA interference impaired viral protein accumulation. Taken together, these findings indicate that autophagy is actively involved in influenza A virus replication.


Pathology of neonatal calf diarrhea induced by a coronavirus-like agent.


title: How should front-line general practitioners use personal protective equipment (PPE)?

text: The COVID-19 outbreak continues to evolve with the number of cases increasing in Malaysia, placing a significant burden on general practitioners (GPs) to assess and manage suspected cases. GPs must be well equipped with knowledge to set up their clinics, use Personal Protective Equipment (PPE) appropriately, adopt standard protocols on triaging and referrals, as well as educate patients about PPE. The correct use of PPE will help GPs balance between personal safety and appropriate levels of public concern.


title: Secondary hemophagocytic lymphohistiocytosis in zoonoses. A systematic review.

text: BACKGROUND Hemophagocytic lymphohistiocytosis (HLH) is a rare syndrome that is often fatal despite treatment. It is caused by a dysregulation in natural killer T-cell function, resulting in activation and proliferation of histiocytes with uncontrolled hemophagocytosis and cytokines overproduction. The syndrome is characterized by fever, hepatosplenomegaly, cytopenias, liver dysfunction, and hyperferritinemia. HLH can be either primary, with a genetic aetiology, or secondary, associated with malignancies, autoimmune diseases, or infections. AIM To focus on secondary HLH complicating zoonotic diseases. MATERIALS AND METHODS PubMed search of human cases of HLH occurring during zoonotic diseases was performed combining the terms (haemophagocytic or haemophagocytosis or hemophagocytosis or hemophagocytic or erythrophagocytosis or macrophage activation syndrome) with each one of the etiological agents of zoonoses. RESULTS Among bacterial diseases, most papers reported cases occurring during brucellosis, rickettsial diseases and Q fever. Regarding viral diseases, most of the cases were reported in patients with avian influenza A subtype H5N1. Among the protozoan zoonoses, most of the cases were reported in patients with visceral leishmaniasis. Regarding zoonotic fungi, most of the cases were reported in AIDS patient with histoplasmosis. No cases of secondary HLH were reported in patient with zoonotic helminthes. CONCLUSIONS Zoonotic diseases are an important cause of HLH. Secondary HLH can delay the correct diagnosis of the zoonotic disease, and can contribute to an adverse outcome.


title: [Spread of infectious agents through refuse by domestic, community and field parasites with special reference to human health].

text: The accumulation of refuse in urban and agricultural areas provides parasitic vermin with new feeding sources and also creates for them partly entirely new biotopes. Vermin transmit both mechanically and biologically the most varied species of pathogens to man, domestic animals and work animals. With respect to the spread of pathogens by vermin via the refuse route, we must distinguish between vermin infesting either the human body, the houses, communities or our environment. Among the human vermin and house species, cockroaches, house gnats and house flies are the most important genera for the spread of pathogens via refuse. Cockroaches transmit bacteria (e.g. Salmonella), viruses (e.g. enterovirus, rota and corona viruses), fungi (e.g. Trichophyton and Candida) and worm eggs (e.g. Ascaris lumbricoides). House flies and house gnats take up all infection carriers from refuse and transmit the pathogens, as a rule, purely mechanically. Rats and mice are the most important species among community vermin. These rodents act both as mechanical and biological vehicles of infectious diseases. A case in point is lymphocytic chorio-meningitis and three different types of hemorrhagic fever in man. Among the vermin infesting our environment, the diptera are the most important carriers. Gnats, especially, often act as intermediate hosts (biological transmission) and, as a result, represent an inexhaustible reservoir of pathogen transmitters. More than 50 diseases caused by arbovirus in man and animal are known. Beside the viruses, the field diptera transmit bacteria (e.g. Rickettsiosis) and protozoa (e.g. Leishmaniasis, Trypanosomiasis).(ABSTRACT TRUNCATED AT 250 WORDS)


Non-contact infrared thermal imagers may still be useful for mass fever screening.


title: Pattern of Colon Cancer Lymph Node Metastases in Patients Undergoing Central Mesocolic Lymph Node Excision: A Systematic Review.

text: BACKGROUND Extended mesocolic lymph node dissection in colon cancer surgery seems to improve oncological outcome. A possible reason might be related to metastases in the central mesocolic lymph nodes. OBJECTIVE The purpose of this study was to describe the pattern of mesocolic lymph node metastases, particularly in central lymph nodes, and the risk of skip, aberrant, and gastrocolic ligament metastases as the argument for performing extended lymph node dissection. DATA SOURCES EMBASE and PubMed were searched using the terms colon or colorectal with sentinel node, lymph node mapping, or skip node; lymph node resection colon; and complete or total and mesocolic excision. STUDY SELECTION Studies describing the risk of metastases in central, skip, aberrant, and gastrocolic ligament lymph node metastases from colon adenocarcinomas in 10 or more patients were included. No languages were excluded. MAIN OUTCOME MEASURES The risk of metastases in the central mesocolic lymph nodes was measured. RESULTS A total of 2052 articles were screened, of which 277 underwent full-text review. The 47 studies fulfilling the inclusion criteria were very heterogeneous, and meta-analyses were not considered appropriate. The risk of central mesocolic lymph node metastases for right-sided cancers varies between 1% and 22%. In sigmoid cancer, the risk is reported in ≤12% of the patients and is associated with advanced T stage. LIMITATIONS The retrospective design and heterogeneity, in terms of definitions of lymph node location, tumor sites, stage, morphology, pathology assessment, and inclusion criteria (selection bias), of the included studies were limitations. Also, anatomic definitions were not uniform. CONCLUSIONS The present literature cannot give a theoretical explanation of a better oncological outcome after extended lymph node dissection. Consensus for a standardization of anatomical definitions and surgical and pathological assessments is warranted for future mapping studies.


title: Establishment of a canine cell line: derivation, characterization, and viral spectrum.

text: A cell line, designated A-72, for virus studies was established from a tumor surgically removed from a female, 8-year-old Golden Retriever dog. Following explant culture, the cells were serially passaged 135 times. The A-72 cells maintained a fibroblastic appearance and, at the 123rd passage, had a population doubling time of approximately 27 hours. Karyotypic analysis of the high passage cells showed the modal 2n chromosome number was 92 to 93. Using starch gel electrophoresis for enzyme characterization, the electrophoretic mobilities of enzymes extracted from A-72 cells were identical with those of canine peritoneal fibroblasts and primary canine kidney cells. The A-72 cells were susceptible to infection with infectious canine hepatitis virus, canine adenovirus type II, canine herpesvirus, canine parainfluenza virus, and canine coronavirus, but were not susceptible to canine distemper virus or the minute virus of canines. These cells have been particularly useful for studies of the fastidious canine coronaviruses, as the commonly used primary canine kidney cells exhibit varied susceptibility to these viruses.


title: Antimicrobial and antiparasitic activity of lectins.

text: Antibiotic resistance is a major problem in current contemporary medicine and it has become a major concern of the 21st century. New resistance mechanisms developed by microorganisms spread greatly, threatening the ability to treat numerous infectious diseases, and increasing the number of nosocomial infections. Besides the role in immunology and glycobiology where they are used as hemaglutinine and identification of complex carbohydrates and glycoconjugates, lectins proved to mediate diversified biological functions like cytotoxicity, complement activation, cell-to-cell and host-pathogen communications, innate immune response, and cell-to-cell signalling. Recently, great interest has been developed for the research and applications of lectins in agriculture and medicine due to their antiparasitic and antimicrobial potentials. This review focuses on the recent data regarding the antimicrobial and antiparasitic activities of lectins, by presenting the role of lectins in host-pathogen interaction and also the cytotoxic effects on microorganisms and parasites. Identification and characterisation of new lectins with antimicrobial activity could serve as a natural alternative for the treatment of infections caused by antibiotic-resistant microorganisms and parasites.


title: Bovine coronavirus mRNA replication continues throughout persistent infection in cell culture.

text: The existence of viral mRNA replicons was demonstrated in cells infected with the bovine coronavirus by showing a minus-strand counterpart and a corresponding replicative intermediate for each subgenomic mRNA species. mRNA replication is thus a universal property of coronaviruses, since this is now the third coronavirus for which it has been demonstrated. During the acute phase of infection (first 48 h), minus and plus strands accumulated at the same rate initially, but maximal accumulation of minus strands peaked earlier than that for plus strands, indicating that minus- and plus-strand levels are differentially regulated. In addition, packaged (input) mRNAs appeared to serve as templates for their own early replication. mRNA replication continued throughout establishment and maintenance of persistent infection (studied for 120 days), which is consistent with our hypothesis that mRNA replication contributes mechanistically to virus persistence. A replication-defective (potentially interfering) species of RNA existed transiently (beginning at day 2 and ending before day 76 postinfection), but because of its transient nature it cannot be considered essential to the long-term maintenance of virus persistence.


title: Processing of the equine arteritis virus replicase ORF1b protein: identification of cleavage products containing the putative viral polymerase and helicase domains.

text: The replicase open reading frame lb (ORF1b) protein of equine arteritis virus (EAV) is expressed from the viral genome as an ORF1ab fusion protein (345 kDa) by ribosomal frameshifting. Processing of the ORF1b polyprotein was predicted to be mediated by the nsp4 serine protease, the main EAV protease. Several putative cleavage sites for this protease were detected in the ORF1b polyprotein. On the basis of this tentative processing scheme, peptides were selected to raise rabbit antisera that were used to study the processing of the EAV replicase ORF1b polyprotein (158 kDa). In immunoprecipitation and immunoblotting experiments, processing products of 80, 50, 26, and 12 kDa were detected. Of these, the 80-kDa and the 50-kDa proteins contain the putative viral polymerase and helicase domains, respectively. Together, the four cleavage products probably cover the entire ORF1b-encoded region of the EAV replicase, thereby representing the first complete processing scheme of a coronaviruslike ORF1b polyprotein. Pulse-chase analysis revealed that processing of the ORF1b polyprotein is slow and that several large precursor proteins containing both ORF1a- and ORF1b-encoded regions are generated. The localization of ORF1b-specific proteins in the infected cell was studied by immunofluorescence. A perinuclear staining was observed, which suggests association with a membranous compartment.


title: Use of gastrointestinal anastomosis stapler for harvest of gracilis muscle and securing it in the face for facial reanimation: a novel technique.

text: BACKGROUND The primary objective of this study is to report a novel technique that uses the gastrointestinal anastomosis (GIA) stapler for harvesting and securing the gracilis muscle in facial reanimation surgery. METHODS We conducted a retrospective chart review with 18 consecutive patients who underwent gracilis muscle flap transfer with or without the use of a GIA stapler. RESULTS Of 11 operations with the GIA stapler, one patient developed a hematoma (donor site) and another required drainage of an abscess (recipient site). Of 8 operations without the use of the stapler, one patient had total flap failure and three required drainage of an abscess (2 recipient sites and 1 donor site). These differences trended toward improvement but were not statistically different. CONCLUSIONS The use of the GIA stapler is a fast, safe technique. Larger studies are, however, warranted to further examine this novel approach in order to test precisely what factors of increased efficiency occur, the amount of suture pull-through, and overall tension capable of being applied to the secured staple line.


title: Designing a biocontainment unit to care for patients with serious communicable diseases: a consensus statement.

text: In spite of great advances in medicine, serious communicable diseases are a significant threat. Hospitals must be prepared to deal with patients who are infected with pathogens introduced by a bioterrorist act (e.g., smallpox), by a global emerging infectious disease (e.g., avian influenza, viral hemorrhagic fevers), or by a laboratory accident. One approach to hazardous infectious diseases in the hospital setting is a biocontainment patient care unit (BPCU). This article represents the consensus recommendations from a conference of civilian and military professionals involved in the various aspects of BPCUs. The role of these units in overall U.S. preparedness efforts is discussed. Technical issues, including medical care issues (e.g., diagnostic services, unit access); infection control issues (e.g., disinfection, personal protective equipment); facility design, structure, and construction features; and psychosocial and ethical issues, are summarized and addressed in detail in an appendix. The consensus recommendations are presented to standardize the planning, design, construction, and operation of BPCUs as one element of the U.S. preparedness effort.


Controversies and uncertainties in managing SARS.


title: Mycoplasma-induced hydrocephalus in rats and hamsters.

text: Mycoplasma pulmonis, a pathogen of the respiratory tract in rats, was inoculated intracerebrally into neonate rats and hamsters to determine if it would induce lesions in the ependyma. Hydrocephalus was induced in 116 of 120 rats and in 23 of 28 hamsters. The severity of hydrocephalus was greater in the rats than in the hamsters. Hydrocephalus induction occurred only subsequent to inoculation of viable M. pulmonis. At 2 weeks of age, rats became refractory to induction of hydrocephalus. Light microscopy indicated that the hydrocephalus was communicating without an inflammatory response in the ventricles and meninges. Preliminary electron microscopy revealed that amorphous material covered portions of the ependymal surface and that cilia were sometimes matted together. It was suggested that the hydrocephalus was due to ciliary dysfunction or to an imbalance of cerebrospinal fluid secretion and absorption. This M. pulmonis-induced hydrocephalus may be a useful model for elucidating the pathogenesis of certain types of congenital hydrocephalus in humans.


title: Research gaps in protecting healthcare workers from SARS and other respiratory pathogens: an interdisciplinary, multi-stakeholder, evidence-based approach.

text: OBJECTIVE To identify priorities for further research in protecting healthcare workers (HCWs) from severe acute respiratory syndrome (SARS) and other respiratory pathogens by summarizing the basic science of infectious bioaerosols and the efficacy of facial protective equipment; the organizational, environmental, and individual factors that influence the success of infection control and occupational health programs; and factors identified by HCWs as important. METHOD An extensive literature review was conducted and 15 focus groups held, mostly with frontline HCWs in Toronto. Critical gaps in knowledge were identified and prioritized. RESULTS Highest priority was given to organizational factors that create a climate of safety. Other priority areas included understanding aerosolization risks and practical measures to control bioaerosols at the source. CONCLUSIONS Further research is warranted to improve safety climate in health care and, specifically, to provide greater protection against respiratory pathogens.