title: In vitro analysis of a hammerhead ribozyme targeted to infectious bronchitis virus nucleocapsid mRNA.

text: Hammerhead ribozymes are catalytic RNA molecules that specifically cleave a target RNA molecule. Herein, we report the design, synthesis, and in vitro analysis of a hammerhead ribozyme targeted to the infectious bronchitis virus (IBV) nucleocapsid mRNA. At a concentration of 0.5 or 10 microM, the ribozyme, designated IBV-N-Rz, effectively cleaved target RNAs in trans (37 C, 10 mM MgCl2, 50 mM Tris). Cleavage products were visualized by agarose gel analysis. The time course of the ribozyme reaction was monitored by agarose gel analysis and relative quantitative reverse transcription-polymerase chain reaction. The amount of target RNA continually declined over a 5-hr period, indicating that the ribozyme was truly catalytic. Although stability and delivery problems must be overcome, a hammerhead ribozyme targeted to the IBV nucleocapsid mRNA most likely has antiviral activity and may be an effective therapeutic/prophylactic reagent in the future.


title: Ministerial burnout: motivation and renewal for mission.

text: Roman Catholic priests are at high risk for stressors, burnout, and other emotional problems due to aging, role confusion, lack of support, changes in occupational focus, and ideological questions (Fruehle, Gautier, Bendyna, 2000; Hamel, 2000; Sammon, Reznikoff, & Gersinger, 1985; USCCB, 1982; USCCB, 2000). The business theories of Organizational Citizen Behavior and Survivor's Syndrome provide organizational explanations for factors contributing to the lack of motivation among priests. The techniques in Motivational Interviewing provide tools for religious leaders to employ when addressing the lack of motivation with individual priests. The article provides recommendations for seminaries and priests' programs of ongoing formation in addressing the issues of burnout, low-morale, and the lack of motivation.


docid: mhinfmal

title: Mouse hepatitis virus does not induce Beta interferon synthesis and does not inhibit its induction by double-stranded RNA.

text: Mouse hepatitis virus (MHV) does not induce interferon (IFN) production in fibroblasts or bone marrow-derived dendritic cells. In this report, we show that the essential IFN-beta transcription factors NF-kappaB and IFN regulatory factor 3 are not activated for nuclear translocation and gene induction during infection. However, MHV was unable to inhibit the activation of these factors and subsequent IFN-beta production induced by poly(I:C). Further, MHV infection did not inhibit IFN-beta production mediated by known host pattern recognition receptors (PRRs) (RIG-I, Mda-5, and TLR3). These results are consistent with the notion that double-stranded RNA, produced during MHV infection, is not accessible to cellular PRRs.


On the Front Lines of Primary Care during the Coronavirus Pandemic Shifting from office visits to telephone triage, telemedicine.


title: The worldwide air transportation network: Anomalous centrality, community structure, and cities' global roles.

text: We analyze the global structure of the worldwide air transportation network, a critical infrastructure with an enormous impact on local, national, and international economies. We find that the worldwide air transportation network is a scale-free small-world network. In contrast to the prediction of scale-free network models, however, we find that the most connected cities are not necessarily the most central, resulting in anomalous values of the centrality. We demonstrate that these anomalies arise because of the multicommunity structure of the network. We identify the communities in the air transportation network and show that the community structure cannot be explained solely based on geographical constraints and that geopolitical considerations have to be taken into account. We identify each city's global role based on its pattern of intercommunity and intracommunity connections, which enables us to obtain scale-specific representations of the network.


docid: lrxskcnp

title: Radiographic-clinical correlation in severe acute respiratory syndrome: study of 1373 patients in Hong Kong.

text: PURPOSE To retrospectively analyze serial chest radiographs in all patients with severe acute respiratory syndrome (SARS) in Hong Kong for temporal changes and differences between patients who died and those who were discharged from the hospital and to compare radiographic and clinical parameters. MATERIALS AND METHODS This retrospective study had ethics review board endorsement, and the need for informed consent was waived. Selected serial chest radiographs obtained from the time of presentation until discharge or death in 1373 patients with laboratory-confirmed SARS were scored. Scoring was based on the area and location of lung opacification on radiographs obtained at each of five milestones (presentation, beginning of ribavirin therapy, beginning of corticosteroid therapy, time of most severe radiographic appearance of disease, and before discharge or death). Extents of lung opacification at these five milestones were compared between patients who died and those who survived (by using a repeated-measures analysis of variance model), and the temporal trend of the radiographic-clinical parameters was analyzed (by using Cochran-Armitage trend testing, Kendall tau correlation coefficients, and descriptive graphic analysis). RESULTS The final cohort consisted of 1373 patients (1212 of whom [485 male and 727 female patients; mean age, 38.4 years] survived and 161 of whom [84 male and 77 female patients; mean age, 63.0 years] died). Among survivors, older patients had more extensive radiographic changes than younger ones. However, among patients who died, older patients had less extensive radiographic opacification at the worst stage of disease and just before death than did younger patients. Despite a higher mortality risk for male patients, both sexes in the same outcome group had similar radiographic findings. For both outcome groups, the rate of radiographic progression was similar for the first 11 days but diverged afterwards. The extent of opacification increased by approximately one zone every 4-5 days for the initial 11 days. Radiographic scores correlated with the ratio of PaO2 to the fraction of inspired oxygen, lymphocyte count, lactate dehydrogenase level, and neutrophil count at each milestone and in terms of changes between milestones (P < .01 for all correlation coefficients, except for radiographic score and neutrophil count between the first two milestones). CONCLUSION The initial extent of radiographic opacification may be useful for prognostic prediction. Radiographic progression correlates well with that of important clinical and laboratory parameters and may be used as an objective prognostic indicator early in SARS.


title: Human coronavirus NL63 employs the severe acute respiratory syndrome coronavirus receptor for cellular entry.

text: Coronavirus (CoV) infection of humans is usually not associated with severe disease. However, discovery of the severe acute respiratory syndrome (SARS) CoV revealed that highly pathogenic human CoVs (HCoVs) can evolve. The identification and characterization of new HCoVs is, therefore, an important task. Recently, a HCoV termed NL63 was discovered in patients with respiratory tract illness. Here, cell tropism and receptor usage of HCoV-NL63 were analyzed. The NL63 spike (S) protein mediated infection of different target cells compared with the closely related 229E-S protein but facilitated entry into cells known to be permissive to SARS-CoV-S-driven infection. An analysis of receptor engagement revealed that NL63-S binds angiotensin-converting enzyme (ACE) 2, the receptor for SARS-CoV, and HCoV-NL63 uses ACE2 as a receptor for infection of target cells. Potent neutralizing activity directed against NL63- but not 229E-S protein was detected in virtually all sera from patients 8 years of age or older, suggesting that HCoV-NL63 infection of humans is common and usually acquired during childhood. Here, we show that SARS-CoV shares its receptor ACE2 with HCoV-NL63. Because the two viruses differ dramatically in their ability to induce disease, analysis of HCoV-NL63 might unravel pathogenicity factors in SARS-CoV. The frequent HCoV-NL63 infection of humans suggests that highly pathogenic variants have ample opportunity to evolve, underlining the need for vaccines against HCoVs.


title: Severe acute respiratory syndrome (SARS) S protein production in plants: development of recombinant vaccine.

text: In view of a recent spread of severe acute respiratory syndrome (SARS), there is a high demand for production of a vaccine to prevent this disease. Recent studies indicate that SARS-coronavirus (CoV) spike protein (S protein) and its truncated fragments are considered the best candidates for generation of the recombinant vaccine. Toward the development of a safe, effective, and inexpensive vaccine candidate, we have expressed the N-terminal fragment of SARS-CoV S protein (S1) in tomato and low-nicotine tobacco plants. Incorporation of the S1 fragment into plant genomes as well as its transcription was confirmed by PCR and RT-PCR analyses. High levels of expression of recombinant S1 protein were observed in several transgenic lines by Western blot analysis using specific antibodies. Plant-derived antigen was evaluated to induce the systemic and mucosal immune responses in mice. Mice showed significantly increased levels of SARS-CoV-specific IgA after oral ingestion of tomato fruits expressing S1 protein. Sera of mice parenterally primed with tobacco-derived S1 protein revealed the presence of SARS-CoV-specific IgG as detected by Western blot and ELISA analysis.


title: Fatal ricin toxicosis in a puppy confirmed by liquid chromatography/mass spectrometry when using ricinine as a marker.

text: Ricin, a lectin from the castor bean plant (Ricinis communis), is considered one of the most potent plant toxins. Ingestion of masticated seeds results in high morbidity, with vomiting and watery to hemorrhagic diarrhea. The prognosis varies with the number of seeds ingested, the degree of mastication, individual susceptibility, and the delay in treatment. Low mortality restricts assessment of histologic lesions, and the literature on toxicologic analysis for ricin is limited. This report describes a fatal case of castor bean ingestion in a 12-week-old Mastiff puppy, with confirmation of ricin exposure through detection of the biomarker ricinine by liquid chromatography/mass spectrometry (LC/MS). Despite supportive therapy, the puppy died several hours after presentation for acute vomiting, diarrhea, and lethargy. At necropsy, a segment of jejunum and mesenteric lymph nodes were congested. When the owner reported the presence of castor beans in the dog's feces, selected formalin-fixed and unfixed tissues were submitted for diagnostic evaluation. Histopathologic findings included superficial necrotizing enteritis of the jejunum and occasional, random foci of coagulative necrosis in the liver. The alkaloid ricinine was detected in gastric content by using a newly developed LC/MS method. This confirmation of exposure is important in the diagnosis of ricin toxicosis, because ingestion of castor beans is not always fatal, histologic lesions are nonspecific, and the degree of mastication can influence the effective dose of ricin.


title: Clinical implications of respiratory virus infections in solid organ transplant recipients: a prospective study.

text: BACKGROUND There is limited information about clinical consequences of respiratory virus infections (RVI) in solid organ transplant recipients. No prospective epidemiological study has been published previously. METHODS We selected a cohort of 152 transplant recipients (cardiac, hepatic and renal transplant recipients). Median time from transplantation was 17 months (range 1-50). They were prospectively followed-up for RVI during 7 months (October to April). Clinical and microbiological evaluation (cell culture, shell vial and polymerase chain reaction technique) of each RVI episode was made. RESULTS We detected 81 RVI (0.91 episodes/patient/year). Complications were detected in 15/81 episodes (18.5%): acute bronchitis (10 cases), pneumonia (three cases; 3.7% of RVI episodes) and bacterial sinusitis (2 cases). In 4 of 81 episodes (5%), patients needed hospitalization. A respiratory virus was isolated in 17 of 68 nasopharyngeal samples (six respiratory syncytial virus, six influenza, four picornavirus, one adenovirus). Fever presented an 83% positive predictive value for the diagnosis of influenza virus infection among those with a positive microbiological isolation. There were no episodes of acute rejection coincidentally with RVI. Only 54% of the subjects had been previously vaccinated against influenza. CONCLUSIONS Incidence of RVI among solid organ transplant recipients is similar to general population but complications are higher. A relationship between RVI and rejection was not detected. The rate of influenza vaccination was lower than expected. The presence of fever in a transplant recipient with RVI strongly suggests influenza infection.


title: Cuba's Pedro Kourí Tropical Medicine Institute:Battling COVID-19 One Study, One Test, One Patient at a Time.

text: This MEDICC Review roundtable brings you specialists from Havana's Pedro Kourí Tropical Medicine Institute (IPK), who are working directly with testing, research and patient care during the COVID-19 pandemic. Founded in 1937 by its namesake, the Institute has gained considerable worldwide prestige. Today, it is a PAHO-WHO Collaborating Center for the Study of Dengue and Its Vector, and for the Elimination of Tuberculosis. Its main role within Cuba's health system is as the national reference center for prevention, control, management and elimination of infectious diseases, including epidemics. Its 479 workers staff 32 departments, including laboratories, research and teach-ing facilities, a hospital and isolation center. The IPK's hospital treats later-stage AIDS patients, while the Institute is the nation-al reference center for attention to all HIV-positive patients and maintains the national HIV/AIDS registry, as well as registries for other infectious diseases. The institution was responsible for training the Cuban doctors who served in West Africa during the 2014-2016 Ebola outbreaks and for those going abroad to assist in the COVID-19 response today, and its professionals offer an internationally-recognized biennial course on dengue.


title: Detection and differentiation of yellow head complex viruses using monoclonal antibodies.

text: Three monoclonal antibodies (MAbs) raised against pathogenic yellow head virus (YHV) from Thailand were tested against tissues of shrimp from Thailand, Australia, Ecuador and India that were purported to be infected with yellow head complex viruses. MAbs V-3-2B and Y-18 were specific to gp116 and gp64 envelope proteins, respectively, while Y-19 was specific to a 20 kDa putative nucleoprotein p20. As a preliminary step, the site of reactivity of the 3 MAbs in YHV was determined by immuno-electron microscopy using ultra-thin sections of YHV-infected shrimp tissue and negatively stained, semi-purified YHV particles. As expected, MAb Y-19 reacted with viral nucleocapsids in ultra-thin sections but not with negatively stained, whole virions; MAb V-3-2B did react with negatively stained, whole virions, but not with virions or nucleocapsids in ultra-thin sections. Unexpectedly, MAb Y-18 did not react with whole or sectioned virions. By immunohistochemistry, MAbs Y-19 and Y-18 reacted with Penaeus monodon tissues infected with either YHV or with gill-associated virus (GAV) from Australia, while MAb V-3-2B reacted with YHV only. In addition, all the YHV and GAV tissue samples gave positive in situ hybridization reactions with a cDNA probe specific to the ORF1b gene of YHV. They also gave expected differential RT-PCR results for YHV and GAV. By contrast, 2 natural Thai shrimp specimens with no gross signs of disease gave similar immunohistochemical reactions and RT-PCR reactions to GAV. However, sequencing of their RT-PCR products showed that they shared 92.7% identity with GAV, but only 79.0% identity with YHV. Although specimens from Ecuador and India displayed histopathology suggestive of YHV infection, they gave negative immunohistochemical reactions with all 3 Mabs, and negative in situ hybridization results. Additional work is required to determine whether a virus from the yellow head complex was responsible for their observed histopathology. These data show that the 3 YHV MAbs could be used in diagnostic situations to differentiate some viruses in the yellow head virus complex.


title: Chemokine regulation of inflammation during acute viral infection.

text: PURPOSE OF REVIEW Chemokines are important inflammatory mediators, and regulate disease due to viral infection. This article will discuss scientific papers published primarily since June 2002 that have introduced new concepts in how chemokines regulate the inflammatory response to specific viruses. RECENT FINDINGS Acute respiratory viruses commonly induce inflammatory chemokines such as CCL3 (also known as macrophage inflammatory protein-1alpha) and CCL5 (RANTES), which can amplify inflammatory responses leading to immunopathology. Where single agent therapy fails, combination antiviral and anti-CCL3 treatment is synergistic and able to prevent mortality in mice infected with the highly lethal pneumonia virus of mice. Human herpesvirus-6 also induces production of CCL3 and CCL5, which are able to block HIV-1 replication in coinfected human lymphoid tissue. On this basis, Margolis has proposed a new and general approach to the treatment and prevention of infection by viral pathogens. SUMMARY Inflammatory chemokines play both beneficial and harmful roles in infectious diseases caused by viruses. Blocking them or using them as immunomodulators, depending on the virus, may be rational approaches to treatment or prevention of disease. With regard to blockade, combination antiviral/antichemokine therapy is a new strategy worth considering as a general therapeutic approach to viral infections, including severe acute respiratory syndrome (SARS). With regard to immunomodulation, use of weak or attenuated viruses to skew the local cytokine network to a configuration able to inhibit a pathogen is a new and interesting concept, but is fraught with important safety issues. Identifying master chemokines to target or exploit in human viral infection is a major opportunity and challenge for clinical immunologists.


title: SARS--lessons from two Singapore General Hospital cases.

text: Two cases of Severe Acute Respiratory Syndrome (SARS) that occurred in Singapore General Hospital are described. The lessons learnt are outlined.


title: Psychiatric complications in patients with severe acute respiratory syndrome (SARS) during the acute treatment phase: a series of 10 cases.

text: In February 2003, Hong Kong was hit by a community-wide outbreak of severe acute respiratory syndrome (SARS). During the period of the outbreak, 10 patients with SARS with psychiatric complications were referred to our Consultation and Liaison Psychiatry Team for assessment and management. We found that both the direct and indirect effects of SARS such as symptom severity, total isolation during treatment and administration of steroid were probable causes of psychiatric complications. In this paper, we report on the nature of their psychiatric problems, challenges to management and psychiatric treatment strategies used during the acute phase.


title: Encountering COVID-19 as Endocrinologists.

text: The world is entering an era of disaster and chaos due to coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2. Since its first emergence in December 2019 in Wuhan, China, COVID-19 has swept through Asia and propagated throughout the world to Europe and North America. As of April 13, 1,773,084 people were infected and 111,652 people had died from COVID-19 globally, and new record levels of infection are being reported every day. Based on the data that have been amassed so far, the primary risk factors for a severe disease course or even mortality from COVID-19 are underlying diseases such as diabetes and hypertension. As the global prevalence of diabetes continues to increase, patients with endocrine diseases such as diabetes mellitus and those who are on long-term corticosteroid therapy due to adrenal insufficiency or hypopituitarism are at risk for a poor prognosis of COVID-19. As endocrinologists, we would like to briefly review the current knowledge about the relationship between COVID-19 and endocrine diseases and to discuss what we can do for the safety and health of our patients with endocrine diseases in this globally threatening situation.


title: Psychological effects of the SARS outbreak in Hong Kong on high-risk health care workers.

text: OBJECTIVE To quantify stress and the psychological impact of severe acute respiratory syndrome (SARS) on high-risk health care workers (HCWs). METHOD We evaluated 271 HCWs from SARS units and 342 healthy control subjects, using the Perceived Stress Scale (PSS) to assess stress levels and a structured list of putative psychological effects of SARS to assess its psychological effects. Healthy control subjects were balanced for age, sex, education, parenthood, living circumstances, and lack of health care experience. RESULTS Stress levels were raised in both groups (PSS = 18) but were not relatively increased in the HCWs. HCWs reported significantly more positive (94%, n = 256) and more negative psychological effects (89%, n = 241) from SARS than did control subjects. HCWs declared confidence in infection-control measures. CONCLUSIONS In HCWs, adaptive responses to stress and the positive effects of infection control training may be protective in future outbreaks. Elevated stress in the population may be an important indicator of future psychiatric morbidity.


title: Local tumor progression patterns after radiofrequency ablation of colorectal cancer liver metastases.

text: PURPOSE We aimed to evaluate patterns of local tumor progression (LTP) after radiofrequency ablation (RF ablation) of colorectal cancer liver metastases (CRCLM) and to highlight the percentage of LTP not attributable to lesion size or RF ablation procedure-related factors (heat sink or insufficient ablation margin). METHODS CRCLM treated by RF ablation at a single tertiary care center from 2004-2012, with a minimum of six months of postprocedure follow-up, were included in this retrospective study. LTP morphology was classified as focal nodular (<90° of ablation margin), circumferential (>270°), or crescentic (90°-270°). Initial metastasis size, minimum ablation margin size, morphology of LTP, presence of a heat sink, and time to progression were recorded independently by two radiologists. RESULTS Thirty-two of 127 RF ablation treated metastases (25%) with a mean size of 23 mm (standard deviation 12 mm) exhibited LTP. Fifteen of 32 LTPs (47%) were classified as focal nodular, with seven having no procedure-related factor to explain recurrence. Ten of 32 LTPs (31%) were circumferential, with four having no procedure-related factor to explain recurrence. Seven of 32 LTPs (22%) were crescentic, with two having no procedure-related factor to explain recurrence. Of the 13 lesions without any obvious procedure-related reason for LTP, six (46%) were <3 cm in size. CONCLUSION Although LTP in RF ablation treated CRCLM can often be explained by procedure-related factors or size of the lesion, in this study up to six (5%) of the CRCLM we treated showed LTP without any reasonable cause.


Severe acute respiratory syndrome: capture-recapture method should be used to count how many cases of SARS really exist.


And the Oscar goes to . . . Salvarsan.


title: Rapid and sensitive detection of bovine coronavirus and group a bovine rotavirus from fecal samples by using one-step duplex RT-PCR assay.

text: Bovine coronavirus (BCoV) and group A bovine rotavirus (BRV) are two of major causes for neonatal calf diarrhea. In the present study, a one-step duplex RT-PCR was established to detect and differentiate BCoV and group A BRV from fecal samples. The sensitivity of this method for BCoV and group A BRV was 10 PFU/100 µl and 1 PFU/100 µl, respectively. Twenty-eight diarrhea fecal samples were detected with this method, the result showed that 2 samples were identified as co-infected with BCoV and group A BRV, 26 samples were group A BRV positive, and 2 samples were negative. It proved that this method is sensitive for clinical fecal samples and is worth applying to laboratory diagnosis for BCoV and group A BRV.


title: Glycosylation of the bovine coronavirus hemagglutinin protein.

text: The bovine coronavirus hemagglutinin protein gp140 is composed of disulfide-linked subunits of 65 kDa. This protein was further characterized with regard to its glycosylation. The glycosylated subunits of the protein are polypeptides having a molecular mass of 42.5 kDa. Both subunits appear to be processed to the same extent by the addition of N-linked oligosaccharides. Each subunit on the mature virion has 6-7 high mannose and 3-4 complex type carbohydrate chains attached to it.


title: Localization of mouse hepatitis virus open reading frame 1A derived proteins.

text: We have investigated the intracellular localization of proteolytic cleavage products encoded in the 5' portion of mouse hepatitis virus (MHV) gene 1. Immunofluorescent labeling of cells with an antiserum which recognizes p28, the ORF1a N-terminal cleavage product, resulted in widespread somewhat granular cytoplasmic staining, indicating that this protein is widely distributed in the cytoplasm of MHV-infected, but not control uninfected cells. Immunofluorescent staining of infected cells with antisera which recognize the downstream polypeptides, p65, p240 and p290 labeled discrete vesicular perinuclear structures. Double immunofluorescent labeling of BHK cells expressing the MHV receptor (BHK(MHVR1)) and infected with MHV-A59 with a Golgi-specific anti-mannosidase II monoclonal antibody and with antiserum recognizing each of these anti-MHV ORF1a polypeptides, showed that the p240 and p290 polypeptides were localized in discrete vesicular structures that overlapped the Golgi complex. Labeling with antibodies specific for p65 colocalized with the Golgi region, and showed staining of the perinuclear cytoplasm as well. Plasmids containing sequences contained in the first 6.75 kb of ORF1a have been expressed using the coupled vaccinia virus-T7 polymerase system. Immunofluorescent labeling of transfectants with the anti-ORF1a antisera showed patterns of antigen distribution similar to those observed in cells infected with MHV-A59. A deletion analysis with constructs containing only portions of the ORF1a sequence indicated that 303 amino acids containing the first papain-like protease domain (PLP-1) was sufficient to associate this protein with the Golgi.


title: [Severe acute respiratory syndrome (SARS)--present status].

text: Basic facts about SARS are presented, including epidemiology, clinical symptoms and the course of disease, multinational search of etiological factor, diagnostic approaches and others. The role of molecular biology in elucidating structure, features and phylogenetic aspects of SARS coronavirus is underlined.


title: Structure of foot-and-mouth disease virus RNA-dependent RNA polymerase and its complex with a template-primer RNA.

text: Genome replication in picornaviruses is catalyzed by a virally encoded RNA-dependent RNA polymerase, termed 3D. The enzyme performs this operation, together with other viral and probably host proteins, in the cytoplasm of their host cells. The crystal structure of the 3D polymerase of foot-and-mouth disease virus, one of the most important animal pathogens, has been determined unliganded and bound to a template-primer RNA decanucleotide. The enzyme folds in the characteristic fingers, palm and thumb subdomains, with the presence of an NH2-terminal segment that encircles the active site. In the complex, several conserved amino acid side chains bind to the template-primer, likely mediating the initiation of RNA synthesis. The structure provides essential information for studies on RNA replication and the design of antiviral compounds.


title: Signs and lesions of experimental Sendai virus infection in two genetically distinct strains of SCID/beige mice.

text: The pathogenesis of Sendai virus infection was studied in genetically immunodeficient mice of genotype scid/scid.bg/bg (SCID-beige) using C.B-17 SCID-beige mice, a BALB/c-related strain that expresses the same major histocompatibility complex as the Sendai virus-susceptible DBA/2 (H-2d). Mice were inoculated intranasally with isolate 771076 of Sendai virus, then killed at 2-day intervals beginning on day 4 post-inoculation. Clinical signs were evident beginning at 8 to 10 days post-inoculation, and all animals remaining were killed in extremis by 14 to 17 days post-inoculation. Lesions in inoculated mice were confined to the respiratory tract. In the nasal passages, a nonresolving rhinitis, with epithelial hyperplasia/metaplasia occurred. Cranioventral bronchopneumonitis was characterized by marked hyperplasia and necrosis of epithelial cells lining airways and with leukocytic infiltration. At the alveolar level, there was marked hypertrophy and hyperplasia of type II pneumocytes, mobilization of alveolar macrophages, and obliteration of the normal architecture in severely affected areas. Viral antigen was evident beginning at 4 days post-inoculation and persisted in affected areas throughout the duration of the study. Because immunocompetent C57BL/6 mice are known to be genetically resistant to Sendai virus, the susceptibility of C57BL/6 SCID-beige to Sendai virus was then compared to that of C.B-17 SCID-beige mice. In age-matched animals of the two strains, there was no evidence of natural resistance to Sendai virus infection in the immunodeficient C57BL/6 strain compared to the C.B-17 mice. These studies indicate that the genetic differences in susceptibility of two strains of immunocompetent mice to Sendai virus infection are eliminated by expression of the mutations scid and beige.


Infection as a cause of multiple sclerosis.


title: Towards a mechanism of function of the viral ion channel Vpu from HIV-1.

text: Vpu, an integral membrane protein encoded in HIV-1, is implicated in the release of new virus particles from infected cells, presumably mediated by ion channel activity of homo-oligomeric Vpu bundles. Reconstitution of both full length Vpu(1-81) and a short, the transmembrane (TM) domain comprising peptide Vpu(1-32) into bilayers under a constant electric field results in an asymmetric orientation of those channels. For both cases, channel activity with similar kinetics is observed. Channels can open and remain open within a broad series of conductance states even if a small or no electric potential is applied. The mean open time for Vpu peptide channels is voltage-independent. The rate of channel opening shows a biphasic voltage activation, implicating that the gating is influenced by the interaction of the dipole moments of the TM helices with an electric field.


title: Stool viruses, coinfections, and diarrhea in HIV-infected patients. Berlin Diarrhea/Wasting Syndrome Study Group.

text: To examine the prevalence of stool viruses and their role in the pathogenesis of diarrhea in HIV infection, we evaluated biopsies and repeated stool samples of 256 HIV-infected patients undergoing diagnostic endoscopy because of diarrhea (n = 136) or other symptoms (n = 120) for bacterial, protozoal, and viral enteropathogens. In 70% of the patients with diarrhea, at least one potential enteropathogen was detected. Stool virus was detected by electron microscopy in 17% (44 of 256), adenovirus in 6.6% (17 of 256), and coronavirus in 11.3% (29 of 256) of the patients. Adenovirus and coronavirus were detected more frequently in patients with diarrhea than in patients without diarrhea [adenovirus 10% (13 of 136) vs. 3.3% (4 of 120), p = 0.0129; coronavirus 15% (21 of 136) vs. 6.6% (8 of 120), p = 0.0142]. Sixty-one percent of patients harboring stool virus were coinfected by another enteropathogen. Pathogens other than stool virus were detected more frequently in patients harboring adenovirus (82%) than in patients without stool virus (48%, p < 0.025). Adenovirus and coronavirus are frequently detected in stools of HIV- infected patients and may contribute to diarrhea. Adenovirus infection may facilitate the occurrence of other intestinal pathogens. Due to frequent coinfections, detection of stool viruses reduces the rate of diarrhea of unknown origin only by approximately 5%.


title: Probiotics for mother and child.

text: For the survival of humankind, nothing can be as important as the health of a mother and a child. As the world's population grows to more than 6 billion, it might seem ridiculous to suggest that any real threat exists to the human species. Diseases have long ravaged populations, as have wars, poverty, and malnutrition. Life today is no different with new and emerging diseases such as SARS and Mad Cow Disease leaving a trail of concern around the planet. All that being said, the AIDS crisis is threatening humans like no other. In countries such as Botswana, close to half the population of pregnant women is infected. Of great concern, the disease is now prevalent among women and teenage girls, threatening not only their lives but those of their offspring. Efforts to control this spread are quite abysmal, albeit well intentioned. Likewise, the death of a child every 15 seconds from diarrheal disease is not being addressed with the same vigor as SARS, even though the risk of dying from the latter for most people is similar to being struck by lightning. In the end, it is the economy and politics that dictate health spending. Image and perception are everything. While deaths mount among women and children from AIDS and other infections, the potential to intervene with a low-risk natural concept of probiotics seemed all too distant. As evidence mounts of the attributes of certain probiotic strains to treat diarrhea and reduce the risk of urogenital and other diseases, the developing world has failed to embrace it, support its evaluation and implementation, and take it to where it is needed the most. In this review, the case for and against probiotics for urogenital and intestinal infections is presented based on current literature. The story is far from complete, but the potential for improving the health of the mother and child is significant. United Nations and World Health Organization guidelines have now been developed to vanquish the unproved marketing hype products that have given probiotics a bad name. It is now up to science to deliver the remedies, and to society to make sure that only proven products reach the marketplace and the people in most need.


title: Parent artery occlusion for unruptured cerebral aneurysms: the Japanese Registry of Neuroendovascular Therapy (JR-NET) 1 and 2.

text: Parent artery occlusion (PAO) is an alternative to surgical clipping or endovascular endosaccular coil embolization for the management of cerebral aneurysms. Most giant and fusiform aneurysms are not amenable to endosaccular coil embolization due to anatomical considerations, such as a broad-neck. However, majority of reports regarding the safety of PAO are based on case series involving a relatively small number of patients. In the present study, a total of 381 consecutive patients with unruptured cerebral aneurysms who were treated with PAO were extracted from the Japanese Registry of Neuroendovascular Therapy (JR-NET) 1 and JR-NET2 database, which are nationwide surveys conducted by the Japanese Society of Neuroendovascular Therapy. The mean age of the 381 patients was 58.1 years, and 59.3% were female. The aneurysmal location included the vertebral artery (42%) and the cavernous portion of internal carotid artery (32%). The aneurysm size and shape consisted of fusiform (45%), giant (25%), and large (22%). Symptomatic lesions were present in 59.8% of the population. Technical success was achieved in 98.4%. The 30-day morbidity and mortality rates were 3.1% and 1.0%, respectively. The most frequent procedure-related complication was ischemic stroke, which occurred in 12.9% (distal embolism, 6.0%; branch occlusion, 3.9%). The 30-day morbidity and mortality rates related to ischemic strokes were 2.1% and 0.3%, respectively. PAO for unruptured aneurysms is feasible with a high technical success rate. Peri-procedural management of ischemic stroke is the key to enhance the safety of this treatment option.


title: Sputum cytology of patients with severe acute respiratory syndrome (SARS).

text: BACKGROUND Severe acute respiratory syndrome (SARS) is a newly described form of atypical pneumonia linked to a novel coronavirus. AIMS To review the sputum cytology of 15 patients who fulfilled the World Health Organisation clinical criteria for SARS in an attempt to evaluate whether early diagnosis is feasible with routine sputum examination. METHODS All sputum samples from patients with SARS from the four major hospitals in Hong Kong were reviewed; abnormalities were sought in the cellular component, including abnormal numbers and morphology of the component cells compared with those from age matched controls taken over the same period one year ago. RESULTS Fifteen sputum samples from patients were compared with 25 control samples. In the patients with SARS, loose aggregates of macrophages were seen more frequently in the sputum. These macrophages frequently showed morphological changes, such as cytoplasmic foaminess, vacuole formation, and nuclear changes (including multinucleation and a ground glass appearance) when compared with the control samples. CONCLUSIONS The cytological features of SARS are non-specific, but the observation of any of the described features should prompt further investigations, especially in patients with suspicious clinical features.


Pneumonia.


title: Intestinal infection of neonatal dogs with canine coronavirus 1-71: studies by virologic, histologic, histochemical, and immunofluorescent techniques.

text: Enteritis induced in neonatal dogs by oral inoculation of coronavirus 1-71 was studied by virologic, histologic, histochemical, and immunofluorescent antibody methods. The enteritis, accompanied by diarrhea, developed in 4 to 7 days and was characterized by atrophy and fusion of intestinal villi and deepening of crypts, increase in cellularity of the lamina propria, flattening of epithelial cells, and discharge of goblet cells. Intra-epithelial enzyme activities generally were depressed. Specific immunofluorescent antibody reactions were present in villous epithelium of the duodenum starting on postinoculation day 2 and of the ileum on day 4. Epiehtlium of the colon showed no reaction. Virus was present in the feces for 6 to 9 days and was recovered from the small and large intestines from day 2 through day 10. The infection was not fatal and was self limiting; the diarrhea stopped between 1 and 2 weeks. The intestinal mucosa also started recovering by the end of week 1, proceeding caudally from the duodenum.


title: [Evaluation of clinical and laboratory findings of pediatric patients with adenovirus-associated respiratory tract infections].

text: Upper respiratory tract infections caused by adenoviruses present long lasting fever for five days and elevated acute phase reactant levels. They are generally misdiagnosed as bacterial infections and are mistreated with antibiotics. The diagnosis of adenovirus infections mainly depends on direct antigen tests, virus isolation and detection of viral DNA using polymerase chain reaction (PCR). The aim of this study was to evaluate the clinical and laboratory findings of the children diagnosed as adenoviral respiratory tract infection by multiplex PCR (mPCR). A total of 27 children (18 male, 9 female; age range: 1-7 years, mean age: 4.4 years) whose nasopharyngeal swab samples were found positive for adenovirus DNA with a commercial mPCR method (Seeplex® RV15 ACE Detection Kit, Seegene Inc, Korea) were included in the study. The throat cultures of the patients revealed no bacterial pathogens and EBV VCA-IgM antibodies were negative. The clinical and laboratory data of the children with long lasting high fever diagnosed as adenovirus infection were evaluated retrospectively in terms of their complaints on admission, symptoms detected in physical examination, laboratory findings and therapy protocols. The patients were categorized according to hospitalization period (< 3 days or ≥ 3 days) and also according to the symptoms compatible with upper or lower respiratory tract infections. The quantity of clinical symptoms (≤ 2 or > 2) and the presence of upper or lower respiratory tract findings were evaluated if there were a difference by means of hospitalization rate and period. The most common complaint of the patients with adenoviral respiratory diseases was fever (27/27; 100%), and the most common admittance season was april-may-june period (20/27; 74%). The mean temperature was 38.4°C (range: 38-39.8°C) and the fever continued for 1-5 days after hospitalization. The most common physical examination finding was tonsillary hyperemia and hypertrophy (63%), followed by lower respiratory tract disease symptoms (37%), otitis media (14.8%), conjunctivitis (7.4%), and rash (3.7%). Laboratory tests could be performed for 24 cases and 95.8% of them yielded high CRP level, 87.5% high sedimentation rate, 62.5% neutrophilia, 33.4% leukocytosis and 20.8% lymphocytosis. It was noticed that 85.2% (23/27) of the patients were under antibiotic treatment on admission. Twenty-three patients (85.2%) were hospitalized, and the duration of hospitalization was between 1-8 (mean: 3.78) days. When the hospitalization rate was evaluated in terms of different measures, it was found that the rate was 81.8% (18/22) in patients with ≤ 2 symptoms, 100% in patients with > 2 symptoms (5/5); 100% (10/10) in patients with lower respiratory tract disease symptoms; 100% (15/15) in patients with neutrophilia, 88.2% (15/17) in patients with CRP levels of ≥ 2.8 - < 100 mg/L, and 100% (6/6) in patients with CRP levels of ≥ 100 mg/L. Neutrophilia and high CRP levels were found to be the main factors related to the hospitalization rate (p< 0.05). In conclusion, adenoviral etiology should be determined by a rapid and sensitive laboratory method such as mPCR, in cases with tonsillopharyngitis who exhibit leukocytosis, neutrophilia and high CRP levels and no bacterial pathogens in throat culture, in order to prevent unnecessary antibiotic use and hospitalization.


title: A randomized, controlled trial of combination therapy for chronic hepatitis B: comparing pegylated interferon-alpha2b and lamivudine with lamivudine alone.

text: BACKGROUND Conventional interferon and lamivudine monotherapy are unsatisfactory in treating hepatitis B virus (HBV) infection. OBJECTIVE To evaluate the efficacy and safety of pegylated interferon-alpha2b and lamivudine combination therapy for chronic hepatitis B. DESIGN Randomized, controlled, open-label trial. SETTING Outpatient clinic in a referral center. PARTICIPANTS 100 treatment-naive patients with hepatitis B e antigen (HBeAg)-positive chronic hepatitis B and moderately elevated alanine aminotransferase levels. MEASUREMENT The primary end point was sustained virologic response (HBeAg seroconversion and HBV DNA level < 500,000 copies/mL) at 24 weeks after cessation of treatment. INTERVENTION A staggered regimen of combination therapy with pegylated interferon-alpha2b (1.5 microg/kg of body weight per week; maximum, 100 microg) given for 32 weeks plus lamivudine (100 mg daily) given for 52 weeks versus lamivudine (100 mg daily) monotherapy given for 52 weeks. Of the 100 participants, 96% completed treatment and 80% completed post-treatment follow-up. RESULTS The rate of sustained virologic response was 36% for the combination treatment group and 14% for the lamivudine monotherapy group (absolute difference, 22 percentage points [95% CI, 6 to 38 percentage points]). End-of-treatment outcomes showed that, compared with monotherapy, patients receiving combination therapy more often had virologic response (60% vs. 28% [absolute difference, 32 percentage points (CI, 14 to 50 percentage points)]); had more substantial reductions of HBV DNA (3.91 log10 copies/mL vs. 2.83 log10 copies/mL); and less often had lamivudine-resistant mutants (21% vs. 40%). The percentages of patients with normalization of alanine aminotransferase levels and histologic improvement did not differ. Adverse effects, such as transient influenza-like symptoms, alopecia, and local erythematous reactions, were more common with combination therapy. LIMITATIONS This study lacked a double-blind design and was conducted at 1 institution. Because of the staggered pegylated interferon-lamivudine regimen, patients assigned to combination therapy received treatment for 8 weeks longer than those assigned to monotherapy. CONCLUSIONS In patients with HBeAg-positive chronic hepatitis B, staggered combination treatment with pegylated interferon-alpha2b and lamivudine may lead to a higher rate of virologic response than lamivudine monotherapy.


title: Indoor air quality in hospitality venues before and after implementation of a clean indoor air law--Western New York, 2003.

text: Secondhand smoke (SHS) contains more than 50 carcinogens. SHS exposure is responsible for an estimated 3,000 lung cancer deaths and more than 35,000 coronary heart disease deaths among never smokers in the United States each year, and for lower respiratory infections, asthma, sudden infant death syndrome, and chronic ear infections among children. Even short-term exposures to SHS, such as those that might be experienced by a patron in a restaurant or bar that allows smoking, can increase the risk of experiencing an acute cardiovascular event. Although population-based data indicate declining SHS exposure in the United States over time, SHS exposure remains a common but preventable public health hazard. Policies requiring smoke-free environments are the most effective method of reducing SHS exposure. Effective July 24, 2003, New York implemented a comprehensive state law requiring almost all indoor workplaces and public places (e.g., restaurants, bars, and other hospitality venues) to be smoke-free. This report describes an assessment of changes in indoor air quality that occurred in 20 hospitality venues in western New York where smoking or indirect SHS exposure from an adjoining room was observed at baseline. The findings indicate that, on average, levels of respirable suspended particles (RSPs), an accepted marker for SHS levels, decreased 84% in these venues after the law took effect. Comprehensive clean indoor air policies can rapidly and effectively reduce SHS exposure in hospitality venues.


title: Prednisolone at anti-inflammatory or immunosuppressive dosages in conjunction with doxycycline does not potentiate the severity of Rickettsia rickettsii infection in dogs.

text: Dogs were experimentally inoculated with Rickettsia rickettsii to determine if anti-inflammatory or immunosuppressive dosages of prednisolone, when administered in conjunction with an antirickettsial antibiotic (doxycycline), induced therapeutically relevant pathophysiological consequences that ultimately influence disease outcome. Although the duration of rickettsemia was prolonged in dogs receiving immunosuppressive, but not anti-inflammatory, corticosteroids, concurrent administration of doxycycline and corticosteroids conferred no other detected detrimental effects. Treatment with doxycycline or doxycycline in conjunction with prednisolone resulted in decreased R. rickettsii-specific antibody titers; however, examination of appropriately timed acute- and convalescent-phase serum samples would have facilitated an accurate diagnosis of Rocky Mountain spotted fever (RMSF) in all 16 dogs. We conclude that the concurrent use of anti-inflammatory or immunosuppressive doses of prednisolone in conjunction with doxycycline, early in the course of experimental RMSF, confers no clinically relevant detrimental effects and that additional studies might be indicated to detect possible beneficial effects in cases of severe or potentially fulminant RMSF. However, because the illness induced in these dogs was of mild to moderate severity, the results of this study should definitely not be construed as supporting the safety or efficacy of prednisolone for treatment of severe canine or human RMSF.


title: Virtual screening of novel noncovalent inhibitors for SARS-CoV 3C-like proteinase.

text: The SARS coronavirus 3C-like proteinase is considered as a potential drug design target for the treatment of severe acute respiratory syndrome (SARS). Owing to the lack of available drugs for the treatment of SARS, the discovery of inhibitors for SARS coronavirus 3C-like proteinase that can potentially be optimized as drugs appears to be highly desirable. We have built a "flexible" three-dimensional model for SARS 3C-like proteinase by homology modeling and multicanonical molecular dynamics method and used the model for virtual screening of chemical databases. After Dock procedures, strategies including pharmocophore model, consensus scoring, and "drug-like" filters were applied in order to accelerate the process and improve the success rate of virtual docking screening hit lists. Forty compounds were purchased and tested by HPLC and colorimetric assay against SARS 3C-like proteinase. Three of them including calmidazolium, a well-known antagonist of calmodulin, were found to inhibit the enzyme with an apparent K(i) from 61 to 178 microM. These active compounds and their binding modes provide useful information for understanding the binding sites and for further selective drug design against SARS and other coronavirus.


title: State of the art: public health and passenger ships.

text: BACKGROUND The purpose of this report is to describe issues relevant to public health and to review all passenger ship associated diseases and outbreaks. Moreover, legislation and practices on sanitation and diseases surveillance related to ships internationally were also reviewed. Some relevant historical information about infections on merchant ships is provided as well. MATERIAL AND METHODS The methods used to develop the state-of-the-art report included a scientific literature review and an extensive and thorough search of the websites of organisations and government departments. A considerable effort was made to capitalise on previous experience in the field. In particular, for the literature review, a total of 158 scientific articles were used including 91 full papers and 67 abstracts, 7 guidelines published by the WHO, and 13 guideline documents published by other organisations. Moreover, 5 international conventions relevant to passenger ships were identified. RESULTS At the international level, public health issues related to ships are regulated by the revised International Health Regulations (2005). Other conventions of the International Maritime Organization regulate safety on board ships and waste and ballast water management, while conventions of the International Labour Organization regulates issues related to working conditions on board ships. Guidelines for preventing and controlling public health threats on board ships can be found in seven Guidelines published by the World Health Organization, including the WHO Guide to Ship Sanitation, which provides a framework for policy making and local decision making. The literature review results revealed that the infections/outbreaks that occur on passenger ships include Norovirus, Legionella spp., Salmonella spp., E. coli, Vibrio spp., and influenza A and B virus. The modes of transmission include person to person, waterborne, foodborne, airborne, and vector-borne, and shore excursions are responsible for some outbreaks. CONCLUSIONS The industry (especially the ferry industry) and sanitary organisations can enhance collaboration in order to implement integrated hygiene programmes and prevent the occurrence of communicable diseases aboard passenger ships.


[Diagnosis and treatment recommendation for pediatric coronavirus disease-19].


docid: rlrziurp

title: Temporal relationship between air pollutants and hospital admissions for chronic obstructive pulmonary disease in Hong Kong.

text: AIMS To assess any relationship between the levels of ambient air pollutants and hospital admissions for chronic obstructive pulmonary disease (COPD) in Hong Kong. METHODS A retrospective ecological study was undertaken. Data of daily emergency hospital admissions to 15 major hospitals in Hong Kong for COPD and indices of air pollutants (sulphur dioxide (SO(2)), nitrogen dioxide (NO(2)), ozone (O(3)), particulates with an aerodynamic diameter of <10 microm (PM(10)) and 2.5 microm (PM(2.5))) and meteorological variables from January 2000 to December 2004 were obtained from several government departments. Analysis was performed using generalised additive models with Poisson distribution, adjusted for the effects of time trend, season, other cyclical factors, temperature and humidity. Autocorrelation and overdispersion were corrected. RESULTS Significant associations were found between hospital admissions for COPD with all five air pollutants. Relative risks for admission for every 10 microg/m(3) increase in SO(2), NO(2), O(3), PM(10) and PM(2.5) were 1.007, 1.026, 1.034, 1.024 and 1.031, respectively, at a lag day ranging from lag 0 to cumulative lag 0-5. In a multipollutant model, O(3), SO(2) and PM(2.5) were significantly associated with increased admissions for COPD. SO(2), NO(2) and O(3) had a greater effect on COPD admissions in the cold season (December to March) than during the warm season. CONCLUSION Ambient concentrations of air pollutants have an adverse effect on hospital admissions for COPD in Hong Kong, especially during the winter season. This might be due to indoor exposure to outdoor pollution through open windows as central heating is not required in the mild winter. Measures to improve air quality are urgently needed.


title: [Pathology and immunohistochemistry of thyroid in severe acute respiratory syndrome].

text: OBJECTIVE To study the pathological characteristics of thyroid gland and the changes on hormone content of thyroid cells in severe acute respiratory syndrome (SARS) patients. METHOD Hematoxylin and eosin staining and light microscopy were used to examine the histology of the thyroid tissues from 4 dead SARS patients and 5 healthy thyroid samples used as negative controls. Immunohistochemistry was used, with monoclonal antibodies, to detect the thyroglobulin (TG), calcitonin and parathyroid hormone (PTH) in the thyroid glands. RESULTS Deformation, enlargement, and dystrophy of follicular cells in thyroid glands were found in the SARS patients. The normal follicular epithelial cells were strongly TG positive, however, the number and intensity of TG positive follicular epithelial cells were significantly lower in the SARS patients. Calcitonin positive cells were found in the normal thyroid glands and not in the thyroid glands of the SARS patients. PTH positive cells were seen in the normal thyroid glands and those of the SARS patients with a slightly weaker intensity of reaction, however, with significant difference in IOD (P < 0.01) and without significant difference in mean slight absorption MOD (P > 0.05). CONCLUSION The thyroid tissue structure and morphology in SARS patients were significantly changed, involving both follicular epithelial cells and parafollicular cells; which may imply that the hormonal production of follicular epithelial cells and the parafollicular cells were affected.


docid: ctuibzsl

title: SYNTHESIS AND AMINOPEPTIDASE N INHIBITING ACTIVITY OF 3-(NITROPHENOXYMETHYL)-[1,3,2]DIOXABOROLAN-2-OLS AND THEIR OPEN ANALOGUES.

text: Aminopeptidase N (APN) represents a class of zinc metallopeptidases with broad substrate specifity. This enzyme is involved in control of angioneogenesis in cancer and microvascular conditions. It also serves as a superficial cellular receptor that enables attachment of some viruses including coronaviruses to the host cell. APN takes part also in metabolism of some important neuropeptides. That is why APN can be a promising therapeutic target and compounds which influence its activity interesting potential drugs. Here, synthesis of compounds which in most contain 3-phenoxypropan-1,2 diol moiety and evaluation of their inhibition activity against APN is described. 4-[1-, 2- and 3-(Nitrophenoxymethyl)]-[1,3,2]dioxaborolan-2-ols are novel compounds which have never been previously reported in the literature. 3-(Aminophenoxy)propyl-1,2-diols revealed greater activity than both 3-(nitrophenoxy)propyl-1,2-diols and 3-(nitrophenoxymethyl)-[1,3,2]dioxaborolan-2-ols. A QSAR study revealed a linear correlation between lipophilicity and inhibition activity.


title: [Ultrastructural characteristics of SARS associated virus in infected cells].

text: OBJECTIVE Electron microscopical study of infected cells to identify the pathogenic agent of SARS. METHODS Vero E6 cells infected with lung autopsy samples or nasopharyngeal swabs from SARS patients of Beijing and Guangzhou were inoculated. The supernatant and cultured cells exhibiting identifiable cytopathic effect (CPE) were prepared for electron microscopic study. RESULTS Examination of CPE cells on thin-section revealed characteristic coronavirus particles within the cisternae of endoplasmic reticulum, Golgi apparatus, vesicles and extracellular space. They were mainly spherical or oval in shape, annular or dense, about 80 nm in diameter. Negative-stain electron microscopy identified coronavirus particles in culture supernatant, 80 - 120 nm in diameter, with club-shaped surface projections. Elongated, rod-, kidney- or other irregular shaped virons with the size of 100 - 200 nm by 60 - 90 nm were also found in the cultured cells infected with the lung samples from the Guangdong patients. Infectious virons entered cells by endocytosis or membrane fusion and released through a budding process. CONCLUSION These data indicate a novel coronavirus as the causative agent of SARS. Most viral particles showed typical characteristics of coronavirus. The potential role of special shape viruses is expected to be further investigated.


title: Monitoring human exposures to upper-room germicidal ultraviolet irradiation.

text: After decades of neglect, the resurgence of tuberculosis in the United States between 1985 and 1992 renewed interest in the use of upper room ultraviolet germicidal irradiation to interrupt the transmission of airborne infections. More recently the bioterrorism threat and the appearance of new pathogens with the potential for airborne spread, such as severe acute respiratory syndrome (SARS), have stimulated installations of upper-room irradiation systems. The objective is to flood the entire volume of a room above 6.5 ft with high intensity ultraviolet germicidal irradiation, while minimizing unintentional irradiance below 6.5 ft to avoid eye and skin irritation. Air exchanges between the upper and lower room result in air disinfection of the occupied space. Designers of these systems have adopted the practice of limiting the maximum lower room irradiance at every point to less than the continuous 8-hour time-weighted average threshold limit value, severely limiting the irradiation intensity in the upper room and thereby reducing one of the two major factors determining germicidal effectiveness, the other being room air mixing. The hypothesis of this study is that eye and skin exposure will be well below the recommended safe dose even when maximum eye-level irradiance levels in the room exceed the 8-hour continuous exposure threshold limit. The method employed was to have subjects wear a small photometer that recorded total ultraviolet dose over the period of exposure while subjects went about their normal routine, and comparing this value with a hypothetical dose calculated from the highest measured eye-level irradiance. The results of the study, based on a limited number of observations, confirmed the hypothesis. Observed doses were one-third to a factor of a hundred or more lower than the doses calculated from maximum eye-level irradiances measurements in the occupants' spaces.


title: Sialic acid is a receptor determinant for infection of cells by avian Infectious bronchitis virus.

text: The importance of sialic acid for infection by avian Infectious bronchitis virus (IBV) has been analysed. Neuraminidase treatment rendered Vero, baby hamster kidney and primary chicken kidney cells resistant to infection by the IBV-Beaudette strain. Sialic acid-dependent infection was also observed with strain M41 of IBV, which infects primary chicken kidney cells but not cells from other species. In comparison with Influenza A virus and Sendai virus, IBV was most sensitive to pre-treatment of cells with neuraminidase. This finding suggests that IBV requires a greater amount of sialic acid on the cell surface to initiate an infection compared with the other two viruses. In previous studies, with respect to the haemagglutinating activity of IBV, it has been shown that the virus preferentially recognizes alpha2,3-linked sialic acid. In agreement with this finding, susceptibility to infection by IBV was connected to the expression of alpha2,3-linked sialic acid as indicated by the reactivity with the lectin Maackia amurensis agglutinin. Here, it is discussed that binding to sialic acid may be used by IBV for primary attachment to the cell surface; tighter binding and subsequent fusion between the viral and the cellular membrane may require interaction with a second receptor.


Hong Kong under WHO spotlight after flu outbreak.


title: Porcine circovirus type 2-associated cerebellar vasculitis in postweaning multisystemic wasting syndrome (PMWS)-affected pigs.

text: Porcine circovirus type 2 (PCV2) is associated with several syndromes in growing pigs, including postweaning multisystemic wasting syndrome and porcine dermatitis and nephropathy syndrome. In the present study, a previously undescribed neurovascular disorder associated with a PCV2 infection is described. Sixteen pigs showed clinical signs of wasting and neurologic deficits. Acute hemorrhages and edema of cerebellar meninges and parenchyma due to a necrotizing vasculitis resulted in degeneration and necrosis of the gray and white matter. Few to numerous PCV2 DNA and antigen-bearing endothelial cells were detected in affected areas of the brain using in situ hybridization and immunohistochemistry. Conventional histochemical stains, as well as the detection of caspase 3 activity and DNA strand breaks by the terminal transferase dUTP nick end labeling assay, showed numerous apoptotic endothelial cells in the vascular lesions observed. Sequencing of various brain-derived PCV2-specific amplicons revealed a strong identity between different isolates and an 89 to 100% identity to previous isolates. The phylogenetic tree showed that there was no clustering of isolates correlating to clinical signs or geographic distribution. This previously undescribed PCV2-associated neurologic disease has features of both postweaning multisystemic wasting syndrome and, to a lesser extent, porcine dermatitis and nephropathy syndrome. The available evidence suggests that direct virus-induced apoptosis of endothelial cells plays a role in the pathogenesis of this unusual PCV2-associated cerebellar vasculitis.


title: Evolution of China's response to HIV/AIDS.

text: Four factors have driven China's response to the HIV/AIDS pandemic: (1) existing government structures and networks of relationships; (2) increasing scientific information; (3) external influences that underscored the potential consequences of an HIV/AIDS pandemic and thus accelerated strategic planning; and (4) increasing political commitment at the highest levels. China's response culminated in legislation to control HIV/AIDS-the AIDS Prevention and Control Regulations. Three major initiatives are being scaled up concurrently. First, the government has prioritised interventions to control the epidemic in injection drug users, sex workers, men who have sex with men, and plasma donors. Second, routine HIV testing is being implemented in populations at high risk of infection. Third, the government is providing treatment for infected individuals. These bold programmes have emerged from a process of gradual and prolonged dialogue and collaboration between officials at every level of government, researchers, service providers, policymakers, and politicians, and have led to decisive action.


Measles virus in the CNS: the role of viral and host factors for the establishment and maintenance of a persistent infection.


title: Detection of coronavirus strain 692 by immune electron microscopy.

text: Utilization of the technique of immune electron microscopy has enabled the detection of a coronavirus in organ culture harvests derived from a washing from an adult with an acute upper respiratory tract illness; convalescent serum was the source of specific antibody.


title: Serologic response of captive coyotes (Canis latrans Say) to canine parvovirus and accompanying profiles of canine coronavirus titers.

text: Fifty-five of 66 (83%) coyote pups from bitches vaccinated against canine parvovirus (CPV) were seropositive for CPV antibodies at birth. The CPV antibody titer in the pups declined with a half-life of 6.7 days until by the 8th week, only two of 41 (5%) pups were seropositive for CPV antibodies. At 8 wk, 41 of the pups were vaccinated against CPV (killed feline origin vaccine), but only one of 37 (3%) was positive for CPV antibodies at 11 wk. The 8-wk-old pups were either too young to respond to the CPV vaccine; they had sufficient undetectable, maternally-derived CPV antibodies to block active immunization; 3 wk was not a sufficient time for an immunological response from the pups; or the vaccine was poorly antigenic. Twenty of the 66 pups (30%) were seropositive for canine coronavirus (CCV) antibodies at birth, and all but three of the 20 were whelped from bitches that were also seropositive for CCV antibodies. Vaccination of females prior to whelping appeared to provide protection to their pups from CPV-induced mortality.


title: Role of endocytosis and low pH in murine hepatitis virus strain A59 cell entry.

text: Infection by the coronavirus mouse hepatitis virus strain A59 (MHV-A59) requires the release of the viral genome by fusion with the respective target membrane of the host cell. Fusion is mediated by the viral S protein. Here, the entry pathway of MHV-A59 into murine fibroblast cells was studied by independent approaches. Infection of cells assessed by plaque reduction assay was strongly inhibited by lysosomotropic compounds and substances that interfere with clathrin-dependent endocytosis, suggesting that MHV-A59 is taken up via endocytosis and delivered to acidic endosomal compartments. Infection was only slightly reduced in the presence of substances inhibiting proteases of endosomal compartments, precluding that the endocytic uptake is required to activate the fusion potential of the S protein by its cleavage. Fluorescence confocal microscopy of labeled MHV-A59 confirmed that virus is taken up via endocytosis. Bright labeling of intracellular compartments suggests their fusion with the viral envelope. No fusion with the plasma membrane was observed at neutral pH conditions. However, when virus was bound to cells and the pH was lowered to 5.0, we observed a strong labeling of the plasma membrane. Electron microscopy revealed low pH triggered conformational alterations of the S ectodomain. Very likely, these alterations are irreversible because low-pH treatment of viruses in the absence of target membranes caused an irreversible loss of the fusion activity. The results imply that endocytosis plays a major role in MHV-A59 infection and the acidic pH of the endosomal compartment triggers a conformational change of the S protein mediating fusion.


title: [Molecular diagnosis and phylogenetic analysis of the first MERS case in Turkey].

text: Coronaviruses (CoV) are enveloped, spherical, single-stranded positive-sense RNA viruses causing mainly respiratory and intestinal infections in animals and humans. Until recently five types of human coronaviruses (HCoV-OC43, HCoV-HKU1, HCoV-NL63, HCoV-229E, SARS-CoV) have been known, however a novel CoV has been identified in 2012 in Saudi Arabia. This virus, namely MERS-CoV (Middle East Respiratory Syndrome Coronavirus), was classified within Coronaviridae family, Coronavirinae sub-family, Betacoronavirus genus, clade C. It causes acute respiratory infections in humans and transmits via respiratory route and close contact between humans. The aim of this study was to present the first MERS case from Turkey identified by molecular methods and the results of viral sequence analysis. A 42-year-old male Turkish citizen who worked as an employee in Jeddah, Kingdom of Saudi Arabia, admitted to hospital with the complaints of fever and malaise on 25-26 September 2014. Since his symptoms went on and got worse, he returned to Turkey, and hospitalized in a hospital's intensive care unit in Hatay on 6th of October with the symptoms of fever, malaise, sweating, cough and respiratory distress. He transferred to a university hospital on 8th of October and died on 11th October. The tracheal aspirate sample obtained before he died was sent to Virology Unit of Reference Laboratories of the Turkish Public Health Institution. Detection of viral RNA was performed by using a commercial real-time PCR kit (hCoV-EMC Real-Time RT-PCR, Fast Track Diagnostics, Luxembourg) targeting the MERS-CoV E protein (upE), ORF1a and ORF1b gene regions. The reference method Superscript III One Step RT-PCR (Invitrogen, USA) recommended by World Health Organization (WHO) was also applied for confirmation. Both of the methods yielded positive results for MERS-CoV RNA. For the amplification of nucleocapsid (N) and RNA-dependent RNA polymerase (RdRp) genes, hemi-nested PCR (Invitrogen, ABD) was conducted, followed by sequence analysis of 204 nucleotide part of N gene. Phylogenetic tree of N gene was obtained with the use of MEGA6 software. N gene was chosen as it comprised a two aminoacid deletion in the corresponding published sequence from the patient treated in London, United Kingdom. There was no nucleotide or aminoacid change in our isolate, namely ANK/1079/2014 when compared with human Betacoronavirus 2c EMC/2012 reference strain found in Genbank database. The target gene regions selected in our study (UpE, ORF1a, ORF1b, N and RdRp) which were also recommended by WHO, shown to have high specificity and sensitivity for the diagnosis and confirmation of MERS-CoV, and also recommended by WHO. The previous studies indicated that, the viral genomes detected in the earliest cases of humans (clade A) are genetically distinct from the others (clade B) which were isolated from dromedary camels and humans. In our study, according to phylogenetic analysis of partial N gene segment, isolate ANK/1079/2014 has taken place within clade A. In conclusion, MERS-CoV appears to have limited circulation in Arabian Peninsula and Middle-Eastern countries, it should be considered in mind that travel-related cases may export the virus outside these regions leading autochtonous infections in the other parts of the world.


title: Feasibility of vascular endothelial growth factor imaging in human atherosclerotic plaque using (89)Zr-bevacizumab positron emission tomography.

text: Intraplaque angiogenesis is associated with the occurrence of atherosclerotic plaque rupture. Cardiovascular molecular imaging can be used for the detection of rupture-prone plaques. Imaging with radiolabeled bevacizumab, a monoclonal anti-vascular endothelial growth factor (VEGF)-A, can depict VEGF levels corresponding to the angiogenic status in tumors. We determined the feasibility of 89Zr-bevacizumab imaging for the detection of VEGF in carotid endarterectomy (CEA) specimens. Five CEA specimens were coincubated with 89Zr-bevacizumab and aspecific 111In-labeled IgG to determine the specificity of bevacizumab accumulation. In 11 CEA specimens, 89Zr-bevacizumab micro-positron emission tomography (PET) was performed following 2 hours of incubation. Specimens were cut in 4 mm wide segments and were stained for VEGF and CD68. In each segment, the mean percent incubation dose per gram of tissue (%Inc/g) and tissue to background ratio were determined. A 10-fold higher accumulation of 89Zr-bevacizumab compared to 111In-IgG uptake was demonstrated by gamma counting. The mean %Inc/ghot spot was 2.2 ± 0.9 with a hot spot to background ratio of 3.6 ± 0.8. There was a significant correlation between the segmental tissue to background uptake ratio and the VEGF score (ρ = .74, p < .001). It is feasible to detect VEGF tissue concentration within CEA specimens using 89Zr-bevacizumab PET. 89Zr-bevacizumab accumulation in plaques is specific and correlates with immunohistochemistry scores.


title: Tissue and cellular distribution of an adhesion molecule in the carcinoembryonic antigen family that serves as a receptor for mouse hepatitis virus.

text: BACKGROUND The receptor for the murine coronavirus mouse hepatitis virus (MHV)-A59, called MHVR or Bgp1a, is a glycoprotein in the carcinoembryonic antigen family of the Ig superfamily. Biliary glycoprotein (Bgp) isoforms play a role in cell adhesion, bile acid transport, and ecto-ATPase activity. MHV-resistant SJL/J mice express a different allele of Bgp1 called Bgp1b. Analysis of the tissue and cellular distribution of Bgp1 proteins can therefore provide new insight on both cellular functions and MHV-A59--induced pathogenesis. EXPERIMENTAL DESIGN Bgp1 expression was analyzed in the digestive, respiratory, endocrine, urinary, and central nervous systems of adult BALB/c and SJL/J mice by immunocytochemistry and immunoelectron microscopy using a monoclonal Ab specific for the N-terminal domain of the Bgp1a proteins and polyclonal rabbit anti-Bgp1, which recognizes both the Bgp1a and Bgp1b proteins. The function of Bgp1 proteins as viral receptors was tested on tissue sections by a virus binding assay. MHV-A59 replication was analyzed by immunocytochemistry. RESULTS Bgp1 expression was observed on membranes of epithelial cells (including hepatocytes, intestinal, endocrine, and respiratory epithelial cells), kidney proximal tubules, and endothelial cells in many tissues. It was usually localized at the apical pole of the cells and, when present, on the brush borders and the cilia. A new direct virus binding assay showed that MHV attachment onto cells correlates with Bgp1 expression. Viral infection was detected in hepatocytes, lymphoid tissue, and the exocrine pancreas but not in endocrine cells, enterocytes, kidney, or respiratory cells. In the central nervous system, no immunolabeling of neurons or glial cells was found with anti-Bgp1 Ab. CONCLUSIONS Bgp1 proteins are present on BALB/c and SJL/J epithelia and endothelia. These glycoproteins might be involved in cell-to-cell contacts, for example between hepatocytes. On BALB/c mice, Bgp1a expression is consistent with the tropism of MHV-A59 for the liver. However, Bgp1a was also expressed on cells that were not infected by MHV-A59.


Emergency Cricothyroidotomy in Tactical Combat Casualty Care.


The health service in a devolved Scotland.


title: Comparison of thiafenantil-xylazine and carfentanil-xylazine for immobilization of gemsbok (Oryx gazella).

text: This study compared immobilization of gemsbok (Oryx gazella) using thiafentanil-xylazine (TX) versus carfentanil-xylazine (CX). Twelve adult gemsboks were divided into two groups, TX and CX. Each group received either 6 mg thiafentanil (0.036 microg/kg (0.032-0.040 microg/kg) and 20 mg xylazine (TX) or 3.6 mg carfentanil (0.021 mg/kg [DOSAGE ERROR CORRECTED] (0.017-0.024 mg/kg) [DOSAGE ERROR CORRECTED] and 20 mg xylazine (CX). Once handled, physiologic measurements were taken every 5 min for 20 min. There were few physiologic differences between the groups, although TX gemsboks were more hypertensive. Quality of induction, quality of recovery, depth of anesthesia, and muscle relaxation were subjectively assessed. Although TX gemsboks showed a trend for more-rapid induction and recovery postantagonist administration, this difference was not statistically significant. Similarly, there was a trend for TX animals to have smoother inductions and improved immobilization quality, but only the quality of induction was significantly different. These trends suggest that TX is preferable to CX when anesthetizing adult gemsboks.


title: De novo initiation of RNA synthesis by the arterivirus RNA-dependent RNA polymerase.

text: All plus-strand RNA viruses encode an RNA-dependent RNA polymerase (RdRp) that functions as the catalytic subunit of the viral replication/transcription complex, directing viral RNA synthesis in concert with other viral proteins and, sometimes, host proteins. RNA synthesis essentially can be initiated by two different mechanisms, de novo initiation and primer-dependent initiation. Most viral RdRps have been identified solely on the basis of comparative sequence analysis, and for many viruses the mechanism of initiation is unknown. In this study, using the family prototype equine arteritis virus (EAV), we address the mechanism of initiation of RNA synthesis in arteriviruses. The RdRp domains of the members of the arterivirus family, which are part of replicase subunit nsp9, were compared to coronavirus RdRps that belong to the same order of Nidovirales, as well as to other RdRps with known initiation mechanisms and three-dimensional structures. We report here the first successful expression and purification of an arterivirus RdRp that is catalytically active in the absence of other viral or cellular proteins. The EAV nsp9/RdRp initiates RNA synthesis by a de novo mechanism on homopolymeric templates in a template-specific manner. In addition, the requirements for initiation of RNA synthesis from the 3' end of the viral genome were studied in vivo using a reverse genetics approach. These studies suggest that the 3'-terminal nucleotides of the EAV genome play a critical role in viral RNA synthesis.


title: Development and evaluation of an ELISA to measure antibody responses to both the nucleocapsid and spike proteins of canine coronavirus.

text: A rapid and reproducible enzyme linked immunosorbent assay (ELISA) was developed for detection of canine coronavirus (CCV) specific antibodies directed to both the nucleocapsid (NC) and the spike (S) proteins. The coating antigen, a methanol-treated, S-protein enriched preparation, was produced by subjecting infected cells to Triton X-114 detergent followed by phase separation. The sensitivity of this assay was determined by following the course of infection in dogs experimentally infected with CCV. The specificity of the antibody response was determined by Western blot analysis and supported the increased magnitude of the ELISA response and the presence of serum neutralizing (SN) antibody. Due to the sensitivity and specificity of the IgG response detected by this assay it can be used to determine both virus exposure and vaccine efficacy.


title: Hepatic T cells and liver tolerance.

text: The T-cell biology of the liver is unlike that of any other organ. The local lymphocyte population is enriched in natural killer (NK) and NKT cells, which might have crucial roles in the recruitment of circulating T cells. A large macrophage population and the efficient trafficking of dendritic cells from sinusoidal blood to lymph promote antigen trapping and T-cell priming, but the local presentation of antigen causes T-cell inactivation, tolerance and apoptosis. These local mechanisms might result from the need to maintain immunological silence to harmless antigenic material in food. The overall bias of intrahepatic T-cell responses towards tolerance might account for the survival of liver allografts and for the persistence of some liver pathogens.


title: Development of a single nucleotide polymorphism DNA microarray for the detection and genotyping of the SARS coronavirus.

text: Severe acute respiratory syndrome (SARS), a disease that spread widely in the world during late 2002 to 2004, severely threatened public health. Although there have been no reported infections since 2004, the extremely pathogenic SARS coronavirus (SARS-CoV), as the causative agent of SARS, has recently been identified in animals, showing the potential for the re-emergence of this disease. Previous studies showed that 27 single nucleotide polymorphism (SNP) mutations among the spike (S) gene of this virus are correlated closely with the SARS pathogenicity and epidemicity. We have developed a SNP DNA microarray in order to detect and genotype these SNPs, and to obtain related information on the pathogenicity and epidemicity of a given strain. The microarray was hybridized with PCR products amplified from cDNAs obtained from different SARS-CoV strains. We were able to detect 24 SNPs and determine the type of a given strain. The hybridization profile showed that 19 samples were detected and genotyped correctly by using our microarray, with 100% accuracy. Our microarray provides a novel method for the detection and epidemiological surveillance of SARS-CoV.


title: Enhancement of plaque formation and cell fusion of an enteropathogenic coronavirus by trypsin treatment.

text: Plaque formation, replication, and related cytopathic functions of the enteropathogenic bovine coronavirus strain L9 in bovine fetal thyroid (BFTy) and bovine fetal brain (BFB) cells were investigated in the presence and absence of trypsin. Plaque formation was enhanced in both cell types. Plaques reached a size with an average diameter of 5 mm within 4 days with trypsin in the overlay, whereas their diameter remained less than 1 mm at this time after plating without trypsin in the overlay. Fusion of both cell types was observed 12 to 18 h after infection when trypsin was present in the medium. Fusion was not observed in infected BFB cell cultures and was rarely observed 48 h after infection of BFTy cells maintained with the trypsin-free medium. The largest polycaryons formed had 15 to 22 nuclei. They then lysed and detached. Cell fusion depended on de novo synthesis of hemagglutinin and infectivity. Fusion from without was not observed. Virus produced under trypsin-enhancing conditions accompanied by cell fusion did not lyse mouse erythrocytes that reacted with L9 coronavirus hemagglutinin. Trypsin-treated, infected BFTy cultures produced coronaviral particles that excluded stain from the envelope confinement. These virions had uniformly shorter surface projections than did the viral forms generated by trypsin-free cell cultures.


title: Inhibition of aminopeptidase N (AP-N) and urokinase-type plasminogen activator (uPA) by zinc suppresses the invasion activity in human urological cancer cells.

text: Zinc is an essential heavy metal and is more abundant in human prostate and kidney than in other tissues. The effects of zinc on the invasion activity of human prostate and renal cancer cell lines, PC-3, LNCaP and SKRC-1, were investigated in vitro using a Transwell cell-culture chamber and were compared with specific protease inhibitors for MMPs, uPA and AP-N, respectively. The invasion activity of PC-3 cells was effectively suppressed by zinc and by all protease inhibitors in a dose-dependent manner. The invasion activity of LNCaP cells was almost unaffected by these inhibitors. In SKRC-1 cells, the invasion activity was strongly suppressed by MP03, although a moderate inhibition by zinc and bestatin was observed. The purified AP-N activity was strongly inhibited by zinc at a concentration similar to that suppressing the invasion activity of PC-3 cells and this inhibition by zinc was apparently competitive. Although the purified uPA activity was also inhibited by zinc, this inhibition was uncompetitive. AP-N was expressed abundantly on the membrane fraction of PC-3 cells among these cells tested, while its expression on the membrane fraction of SKRC-1 cells was weaker than that of PC-3 cells. The expression of uPA was also highest on the membrane fraction of PC-3 cells. These results suggest that AP-N and uPA may be involved in the invasion of human prostate cancer cells and that zinc probably participates in the invasion and metastasis of cancer cells through the regulation of the enzymatic activity of AP-N and uPA in human cancerous prostate.


Follow up of feline immunodeficiency virus-infected cats.


title: Transmission routes of SARS-CoV-2 and protective measures in dental clinics during the COVID-19 pandemic.

text: RESULTS The outbreak of coronavirus disease 2019 (COVID-19) has become a primary challenging public health issue for not only China but also the world. On March 11, 2020, the World Health Organization declared that the pandemic of COVID-19 had become a public health emergency of global concern. As of May 12, 2020 COVID-19 has been reported in over 180 countries/regions, with a total of 4,178,156 confirmed cases and over 280,000 deaths. The risk of cross-infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) might be elevated between dental specialists and patients due to the peculiarity of dental practice. Therefore, detailed and effective infection control measures are imminently needed to prevent nosocomial coronavirus infection. This paper, based on proven effective experience, relevant guidelines, and research, not only presents the fundamental knowledge about the name, etiologic characteristics, epidemiologic characteristics and clinical manifestations of COVID-19, transmission routes and risk points of SARS-CoV-2 in dental clinics but also provides recommended protective measures for dental professionals to interdict the transmission of SARS-CoV-2 in dental clinics. Classification schemes as well as color identification according to the results of the questionnaire survey and temperature measurement in precheck and triages are innovations proposed in this paper. CLINICAL SIGNIFICANCE This paper, based on proven effective experience, relevant guidelines, and research, not only presents the fundamental knowledge about the name, etiologic characteristics, epidemiologic characteristics and clinical manifestations of COVID-19, transmission routes and risk points of SARS-CoV-2 in dental clinics but also provides recommended protective measures for dental professionals to interdict the transmission of SARS-CoV-2 in dental clinics.


title: Stem-loop III in the 5' untranslated region is a cis-acting element in bovine coronavirus defective interfering RNA replication.

text: Higher-order structures in the 5' untranslated region (UTR) of plus-strand RNA viruses are known in many cases to function as cis-acting elements in RNA translation, replication, or transcription. Here we describe evidence supporting the structure and a cis-acting function in defective interfering (DI) RNA replication of stem-loop III, the third of four predicted higher-order structures mapping within the 210-nucleotide (nt) 5' UTR of the 32-kb bovine coronavirus (BCoV) genome. Stem-loop III maps at nt 97 through 116, has a calculated free energy of -9.1 kcal/mol in the positive strand and -3.0 kcal/mol in the negative strand, and has associated with it beginning at nt 100 an open reading frame (ORF) potentially encoding an 8-amino-acid peptide. Stem-loop III is presumed to function in the positive strand, but its strand of action has not been established. Stem-loop III (i) shows phylogenetic conservation among group 2 coronaviruses and appears to have a homolog in coronavirus groups 1 and 3, (ii) has in all coronaviruses for which sequence is known a closely associated short, AUG-initiated intra-5' UTR ORF, (iii) is supported by enzyme structure-probing evidence in BCoV RNA, (iv) must maintain stem integrity for DI RNA replication in BCoV DI RNA, and (v) shows a positive correlation between maintenance of the short ORF and maximal DI RNA accumulation in BCoV DI RNA. These results indicate that stem-loop III in the BCoV 5' UTR is a cis-acting element for DI RNA replication and that its associated intra-5' UTR ORF may function to enhance replication. It is postulated that these two elements function similarly in the virus genome.


title: Combined usage with intraperitoneal and incisional ropivacaine reduces pain severity after laparoscopic cholecystectomy.

text: Postoperative pain is the main obstacle for safely rapid recovery of patients undergoing laparoscopic cholecystectomy (LC). In this study, we systemically evaluated the analgesic efficacy of intraperitoneal and incisional ropivacaine injected at the end of the LC. A total of 160 patients, scheduled for elective LC, were allocated into four groups. Group Sham received intraperitoneal and incisional normal saline (NS). Group IC received incisional ropivacaine and intraperitoneal NS. Group IP received incisional NS and intraperitoneal ropivacaine. Group ICP received intraperitoneal and incisional ropivacaine. At the end of the surgery, ropivacaine was injected into the surgical bed through the right subcostal port and infiltrated at the four ports. Dynamic pain by a visual analogue scale (VAS) and cumulative morphine consumption at 2 h, 6 h, 24 h, and 48 h postoperatively, as well as incidence of side-effects over 48 h after LC was recorded. Compared with those in group Sham, the time of post-anesthesia care unit (PACU) stay, dynamic VAS score (VAS-D) 2 h and 6 h postoperatively, cumulative morphine consumption 6 h and 24 h postoperatively, and incidence of nausea and vomiting 48 h after LC in group IC and ICP were less (P<0.05). Furthermore, intraperitoneal and incisional ropivacaine exerts more powerful analgesic effect than single usage with intraperitoneal or incisional ropivacaine (P<0.05). No patients exhibited signs of local anesthetic toxicity. In conclusion, intraperitoneal and incisional ropivacaine might facilitate PACU transfer and effectively and safely reduce pain intensity after LC.


title: Short-Term Dexamethasone in Sars-CoV-2 Patients.

text: BACKGROUND Dexamethasone, a synthetic glucocorticoid, has anti-inflammatory and immunosuppressive properties. There is a hyperinflammatory response involved in the clinical course of patients with pneumonia due to SARS-CoV-2. To date, there has been no definite therapy for COVID-19. We reviewed the charts of SARS-CoV-2 patients with pneumonia and moderate to severely elevated CRP and worsening hypoxemia who were treated with early, short-term dexamethasone. METHODS We describe a series of 21 patients who tested positive for SARS-CoV-2 and were admitted to The Miriam Hospital in Providence, RI, and were treated with a short course of dexamethasone, either alone or in addition to current investigative therapies. RESULTS CRP levels decreased significantly following the start of dexamethasone from mean initial levels of 129.52 to 40.73 mg/L at time of discharge. 71% percent of the patients were discharged home with a mean length of stay of 7.8 days. None of the patients had escalation of care, leading to mechanical ventilation. Two patients were transferred to inpatient hospice facilities on account of persistent hypoxemia, in line with their documented goals of care. CONCLUSIONS A short course of systemic corticosteroids among inpatients with SARS-CoV-2 with hypoxic respiratory failure was well tolerated, and most patients had improved outcomes. This limited case series may not offer concrete evidence towards the benefit of corticosteroids in COVID-19. However, patients' positive response to short-term corticosteroids demonstrates that they may help blunt the severity of inflammation and prevent a severe hyperinflammatory phase, in turn reducing the length of stay, ICU admissions, and healthcare costs.


title: The role of the NxTAG® respiratory pathogen panel assay and other multiplex platforms in clinical practice.

text: INTRODUCTION The advent of nucleic acid amplification tests has significantly improved the aetiologic diagnosis of respiratory infections. However, multiplex real-time polymerase chain reaction (PCR) can be technologically challenging. Areas covered: This paper reports the results of the main published studies on the NxTAG Respiratory Pathogen Panel (RPP) and discusses the advantages and disadvantages of extensive use of multiplex assays in clinical practice. Expert commentary: Currently available data seem to indicate that routine use of multiplex assays, including NxTAG RPP Assay, should be recommended only when epidemiological data concerning circulation of viruses and bacteria have to be collected. Their use in clinical practice seems debatable. They have limited sensitivity and specificity at least in the identification of some infectious agents or, as in the case of NxTAG RPP, they have not been evaluated in a sufficient number of patients to allow definitive conclusions. In the future, the clinical relevance of multiplex assays, including NxTAG RPP, could significantly increase, mainly because a number of new antiviral agents effective against several respiratory viruses for which no drug is presently available will be marketed. In addition, it is highly likely that the efficiency of multiplex assays will be significantly improved.


title: Isolation of Propionibacterium acnes from a case of placentitis and abortion in a cow.

text: On the basis of the scarcity of reports in the veterinary literature, it appears that Propionibacterium spp. are rarely associated with disease or isolated from cattle tissues. Recently, Propionibacterium spp. has been associated with multifocal abscessation in cattle. This report describes a case of necrosuppurative placentitis and abortion in an adult Holstein cow. Numerous colonies of small, pleomorphic, Gram-positive, rod-shaped bacteria were observed within the fibrin lattice associated with placental lesions and within the fetal atelectatic lung. Propionibacterium acnes was isolated in high numbers from the placenta, fetal lung, and stomach contents. To the authors' knowledge, this is the first report of placentitis associated with propionibacteria in a cow.


Health Research Symposium 2010: improving health and recognising excellence.


title: Detection and estimation of avian infectious bronchitis virus antigen by a novel indirect liquid-phase blocking enzyme-linked immunosorbent assay using chicken and rabbit affinity purified immunoglobulins.

text: An indirect liquid-phase blocking (LPB) enzyme-linked immunosorbent assay (ELISA) using chicken and rabbit affinity purified immunoglobulin G (IgG) has been developed to detect and estimate avian infectious bronchitis virus (IBV) antigen concentration directly in infected allantoic fluid. The method is based on the principle of binding of specific IgG to the test IBV antigen and the assay of unbound IgG on an antigen-coated ELISA plate. The immunoglobulins are chicken N-terminal S2 peplomeric protein-specific IgG isolated by immunoaffinity chromatography on synthetic peptide coupled to CNBr-activated Sepharose 4B or rabbit polyclonal IgG purified from the serum using Protein A Sepharose 4B. The assay detected all tested IBV strains and field isolates propagated in chicken embryos. Signal to noise ratios were calculated from LPB ELISA absorbance units and a diagnostic threshold was established from the signal to noise ratio frequency distribution of samples positive or negative for IBV by virus titration or reverse transcription polymerase chain reaction. The relative sensitivity of the test ranged between 10(5) and 10(6) median egg infectious doses (EID(50)) for chicken IgG and between 10(3) and 10(4) EID(50) for rabbit IgG, depending on the test strain. The assay is simple and takes less than 3 h to perform. It does not require expensive reagents and can be readily adapted to monitor the IBV antigen concentration in allantoic fluids during propagation of vaccine strains or in samples of freeze-dried, live-attenuated IBV vaccines.


title: Severe acute respiratory syndrome-associated coronavirus nucleocapsid protein interacts with Smad3 and modulates transforming growth factor-beta signaling.

text: Severe acute respiratory syndrome (SARS) is an acute infectious disease with significant mortality. A typical clinical feature associated with SARS is pulmonary fibrosis and the associated lung failure. However, the underlying mechanism remains elusive. In this study, we demonstrate that SARS-associated coronavirus (SARS-CoV) nucleocapsid (N) protein potentiates transforming growth factor-beta (TGF-beta)-induced expression of plasminogen activator inhibitor-1 but attenuates Smad3/Smad4-mediated apoptosis of human peripheral lung epithelial HPL1 cells. The promoting effect of N protein on the transcriptional responses of TGF-beta is Smad3-specific. N protein associates with Smad3 and promotes Smad3-p300 complex formation while it interferes with the complex formation between Smad3 and Smad4. These findings provide evidence of a novel mechanism whereby N protein modulates TGF-beta signaling to block apoptosis of SARS-CoV-infected host cells and meanwhile promote tissue fibrosis. Our results reveal a novel mode of Smad3 action in a Smad4-independent manner and may lead to successful strategies for SARS treatment by targeting the TGF-beta signaling molecules.


title: A study in hospital noise - a case from Taiwan.

text: Hospitals are places that allow patients to rest and recover, and therefore must be quiet inside and in the surrounding neighborhood. One medical center was chosen as a sample hospital. This hospital was a tertiary care center during the 2003 outbreak of the severe acute respiratory syndrome (SARS) in Taiwan. The measurement results show that the noise level in the wards and stations was between 50.3 and 68.1 dB which exceeded the suggested hospital ward sound level. The quietest units were the Surgical Intensive Care Unit and recovery rooms with a noise level lower than 50 dB during the night. The higher noise levels were in the hall and pharmacy which were highly populated areas. This study analyzed the causes of this excessive noise and used noise reduction methods. The paired t test was performed and the results showed improvement methods were successful. This study found the noise levels reached 98.5-107.5 dB in power generator rooms and air-conditioning facilities, and suggests employees use ear plugs.