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].


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.


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.


title: Surgical management of symptomatic hydatid liver disease: experience from a Western centre.

text: BACKGROUND Hydatid liver cysts are rare in North America. The objective of this study was to determine the optimal surgical management for hydatid liver cysts treated outside endemic areas. METHODS We reviewed the cases of consecutive patients who underwent management of hydatid liver cysts. Radical liver resections were compared with other types of procedures. Clinical presentation, investigations, perioperative outcomes and longterm follow-up were evaluated. We evaluated disease recurrence using the Kaplan- Meier method. RESULTS Forty patients underwent surgery for hydatid liver cysts. Most patients had single (68%) right-sided (46%) cysts with a median size of 10 cm. Most (83%) underwent liver resection with or without drainage/marsupialization. Radical liver resection was carried out in 60% (19 major, 5 minor). Additional procedures were required in 50% (biliary fistulization 30%, diaphragmatic fistulization 20% or paracaval location/ fusion 8%). Postoperative complications occurred in 48%. The median follow-up was 39 months. The 3-year recurrence-free survival was significantly different between patients who had radical resection and those who had other procedures (100% v. 71%, p = 0.002). CONCLUSION The surgical management of hydatid liver cysts in North America remains rare and challenging and is frequently associated with fistulizing complications. Excellent long-term outcomes are best achieved using principles of radical liver resection that are familiar to North American surgeons.


Hygiene: what and why?


title: Meeting highlights of the European Respiratory Society annual congress 02-06 September 2006, Munich, Germany.

text: In pulmonary medicine, there are two major international meetings held annually: the annual meeting of the American Thoracic Society (ATS), and that of the European Respiratory Society (ERS). The 2006 ERS Annual Congress was held in Munich, Germany. Both meetings provide an annual forum for scientists and clinical investigators from academia, non-university institutions (including private practices) and pharmaceutical companies to share information on many aspects of pulmonary and critical-care medicine, sleep disorders, pulmonary infectious diseases, malignancies of the chest, and numerous aspects of diagnostic and interventional procedures in these fields. With > 5000 abstracts and ~ 17,000 participants, the 2006 ERS-congress outnumbered even the 2006 ATS meeting. Research areas covered all aspects of pulmonary diseases, although this year (as in previous years) obstructive pulmonary diseases such as COPD (chronic obstructive pulmonary disease) and asthma were clearly the main focus.


title: The homeostatic rôle of bronchoconstriction.

text: This article argues in favour of the hypothesis that the homeostatic rôles of bronchoconstriction are to retract the airway tree during expiration, and to assist in the expulsion of mucus from peripheral airways by increasing the velocity of outgoing air. In asthma, this function may be dangerously exaggerated because of the presence of unusually viscous airway secretions and the remodelling of airway walls.


title: Anterior iliopsoas impingement and tendinitis after total hip arthroplasty.

text: Anterior iliopsoas impingement and tendinitis is a poorly understood and likely underrecognized cause of groin pain and functional disability after total hip arthroplasty. The patient history and physical examination findings are usually only suggestive, and the symptoms frequently subtle. The diagnosis may be confirmed by one or more imaging studies, including a cross-table lateral radiograph, computed tomography, magnetic resonance imaging, and ultrasonography, in combination with a confirmatory diagnostic injection. Nonsurgical management may not resolve the problem. Surgical treatment, consisting of release or resection of the iliopsoas tendon, alone or in combination with acetabular revision for an anterior overhanging component, usually provides permanent pain relief.


title: [Analysis on the trend of severe acute respiratory syndrome epidemic in Inner Mongolian Autonomous Region].

text: OBJECTIVE To analyse the severe acute respiratory syndrome (SARS) epidemics in Inner Mongolian Autonomous Region and to provide scientific basis for prevention and control strategies against it. METHODS Data from legal communicable diseases surveillance reporting system was analyzed epidemiologically. RESULTS The first SARS case was reported in Inner Mongolian Autonomous Region on March 27, 2003. Up to May 20, there were 446 cumulative SARS cases in the whole region (with 287 confirmed cases and 159 suspected cases) and 61 cumulative recovered cases had been discharged from the hospitals (56 confirmed cases and 5 suspected cases). Another 131 cases were excluded the original diagnoses of SARS including 10 confirmed cases and 121 suspected cases. 25 confirmed cases died with a mortality rate of 8.7%. Cumulatively, the number of reported cases were distributed in 30 counties in 9 prefectures. Statistical analysis on time sequence of the occurrence of cases showed that majority (67.7% of the total) of the cases concentrated in between April 13 and April 29. The number of cases had started to decrease since April 24 with an average of 5.3 cases per day between May 3 and May 8 and an average of 0.3 cases per day between May 9 and today. CONCLUSIONS SARS epidemics in our region could be divided into three phases. The first phase fell in between March 18 and April 15 with the first case being imported, the number of cases rose sharply, covering 14 counties in 6 prefectures, having a feature of family clustering. The second phase was from April 16 to April 28, with the appearance of secondary infection, having sharp rise of the cases and spreading to 24 counties in 10 prefectures. One of the major features was that hospitals had become the important sources of secondary infection. Finally, the third phase was between April 29 and May 20, with small wave crests of cases, spreading to 38 counties in 10 prefectures with a high proportion of cases with no history of direct contact with diagnosed SARS patients. Thus, no obvious transmission chain was noticed at this phase.


title: Characterization of the expression of the hepatitis C virus F protein.

text: Hepatitis C virus (HCV) is an important human pathogen that affects 170 million people worldwide. The HCV genome is approximately 9.6 kb in length and encodes a polyprotein that is proteolytically cleaved to generate at least 10 mature viral protein products. Recently, a new protein, named F, has been described to be expressed through a ribosomal frameshift within the capsid-encoding sequence, a mechanism unique among members of the family Flavidiridae: Here, expression of the F protein was investigated in an in vitro transcription/translation assay. Its expression in mammalian cells was confirmed using specific recombinant vaccinia viruses; under these conditions, protein expression is dependent on the HCV IRES. The F protein was tagged with firefly luciferase or the Myc epitope to facilitate its identification. Ribosomal frameshifting was dependent on the presence of mutations in the capsid-encoding sequence. No frameshifting was detected in the absence of any mutation. Furthermore, analysis of the F protein in time-course experiments revealed that the protein is very unstable and that its production can be stabilized by the proteasome inhibitor MG132. Finally, indirect immunofluorescence studies have localized the F protein in the cytoplasm, with notable perinuclear detection.


title: Regulation of relative abundance of arterivirus subgenomic mRNAs.

text: The subgenomic (sg) mRNAs of arteriviruses (order Nidovirales) form a 5'- and 3'-coterminal nested set with the viral genome. Their 5' common leader sequence is derived from the genomic 5'-proximal region. Fusion of sg RNA leader and "body" segments involves a discontinuous transcription step. Presumably during minus-strand synthesis, the nascent RNA strand is transferred from one site in the genomic template to another, a process guided by conserved transcription-regulating sequences (TRSs) at these template sites. Subgenomic RNA species are produced in different but constant molar ratios, with the smallest RNAs usually being most abundant. Factors thought to influence sg RNA synthesis are size differences between sg RNA species, differences in sequence context between body TRSs, and the mutual influence (or competition) between strand transfer reactions occurring at different body TRSs. Using an Equine arteritis virus infectious cDNA clone, we investigated how body TRS activity affected sg RNA synthesis from neighboring body TRSs. Flanking sequences were standardized by head-to-tail insertion of several copies of an RNA7 body TRS cassette. A perfect gradient of sg RNA abundance, progressively favoring smaller RNA species, was observed. Disruption of body TRS function by mutagenesis did not have a significant effect on the activity of other TRSs. However, deletion of body TRS-containing regions enhanced synthesis of sg RNAs from upstream TRSs but not of those produced from downstream TRSs. The results of this study provide considerable support for the proposed discontinuous extension of minus-strand RNA synthesis as a crucial step in sg RNA synthesis.


title: Sudden death associated with Clostridium sordellii in captive lions (Panthera leo).

text: In the spring of 2003, a series of sudden deaths in a group of adult lions (Panthera leo) with a previous history of depression, inanition, and lethargy, was investigated. Five animals died within 24 to 36 hours after onset of signs of disease. Serologic screening for viral disease detection was negative, evidence of parasites was not detected, and results of a complete blood count and serum biochemical analysis were within reference intervals in all lions. The most relevant lesions observed were multiple areas of necrosis and hemorrhage in the intestinal outer muscular layer, and cellulitis with an intense bloody edema in the mesenteric and the pericardial fat tissue. On the basis of the fulminant course of the disease, the gross and histologic findings, and the isolation and identification of Clostridium sordellii, a diagnosis of infectious myositis and cellulitis associated with acute clostridiosis was made. To the authors' knowledge, this is the first report of sudden death associated with C. sordellii in felines.


title: Detection and serotyping of human adenoviruses from patients with influenza-like illness in mongolia.

text: Human adenoviruses (HAdVs) are responsible for approximately 5%-10% of acute respiratory infections. The serotypes of commonly detected respiratory HAdV in Asian countries are diverse. However, there are no well-documented reports of circulating HAdV serotypes in Mongolia. Between January 2010 and May 2011, 1,950 influenza-negative samples from patients with influenza-like illness, including eye swabs from patients with eye symptoms, were screened for HAdV, and 40 samples (2.1%) were positive for HAdVs. Among these 40 samples, 31 samples were positive for the hexon gene used in phylogenetic analysis, as determined by PCR. We identified 7 different serotypes. We constructed the phylogenetic trees of HAdV-B7 and HAdV-B3, the 2 most commonly detected serotypes in this study. All detected HAdV-B7 and -B3 Mongolian strains had identical sequences. HAdV-D8, known to be associated with epidemic keratoconjunctivitis (EKC), was detected from nasopharyngeal and eye swabs. There was no difference between the amino acid sequences of the hexon and fiber genes that may affect tissue tropism in Mongolian strains and those in EKC-causing strains.


title: Human parechovirus type 1, 3, 4, 5, and 6 detection in picornavirus cultures.

text: Picornavirus cultures that could not be typed in neutralization assays were analyzed by VP1 reverse transcription-PCR (RT-PCR) and a virus discovery tool (VIDISCA). Human parechoviruses (HPeVs) were frequently identified, among which were the uncommon isolates HPeV-4, HPeV-5, and HPeV-6. The HPeV-5 isolate could be amplified only by VIDISCA and not by VP1 RT-PCR.


title: The 3C-like proteinase of an invertebrate nidovirus links coronavirus and potyvirus homologs.

text: Gill-associated virus (GAV), a positive-stranded RNA virus of prawns, is the prototype of newly recognized taxa (genus Okavirus, family Roniviridae) within the order NIDOVIRALES: In this study, a putative GAV cysteine proteinase (3C-like proteinase [3CL(pro)]), which is predicted to be the key enzyme involved in processing of the GAV replicase polyprotein precursors, pp1a and pp1ab, was characterized. Comparative sequence analysis indicated that, like its coronavirus homologs, 3CL(pro) has a three-domain organization and is flanked by hydrophobic domains. The putative 3CL(pro) domain including flanking regions (pp1a residues 2793 to 3143) was fused to the Escherichia coli maltose-binding protein (MBP) and, when expressed in E. coli, was found to possess N-terminal autoprocessing activity that was not dependent on the presence of the 3CL(pro) C-terminal domain. N-terminal sequence analysis of the processed protein revealed that cleavage occurred at the location (2827)LVTHE downward arrow VRTGN(2836). The trans-processing activity of the purified recombinant 3CL(pro) (pp1a residues 2832 to 3126) was used to identify another cleavage site, (6441)KVNHE downward arrow LYHVA(6450), in the C-terminal pp1ab region. Taken together, the data tentatively identify VxHE downward arrow (L,V) as the substrate consensus sequence for the GAV 3CL(pro). The study revealed that the GAV and potyvirus 3CL(pro)s possess similar substrate specificities which correlate with structural similarities in their respective substrate-binding sites, identified in sequence comparisons. Analysis of the proteolytic activities of MBP-3CL(pro) fusion proteins carrying replacements of putative active-site residues provided evidence that, in contrast to most other 3C/3CL(pro)s but in common with coronavirus 3CL(pro)s, the GAV 3CL(pro) employs a Cys(2968)-His(2879) catalytic dyad. The properties of the GAV 3CL(pro) define a novel RNA virus proteinase variant that bridges the gap between the distantly related chymotrypsin-like cysteine proteinases of coronaviruses and potyviruses.


Linear neutralizing epitopes on the peplomer protein of coronaviruses.


title: Alphavirus minus-strand synthesis and persistence in mouse embryo fibroblasts derived from mice lacking RNase L and protein kinase R.

text: We report our studies to probe the possible role of the host response to double-stranded RNA in cessation of alphavirus minus-strand synthesis. Mouse embryo fibroblasts (MEF) from Mx1-deficient mice that also lack either the protein kinase R (PKR) or the latent RNase L or both PKR and RNase L were screened. In RNase L-deficient but not wild-type or PKR-deficient MEF, there was continuous synthesis of minus-strand templates and the formation of new replication complexes producing viral plus strands. Inhibiting translation caused minus-strand synthesis to stop and a loss of transcription activity of the mature replication complexes. This turnover of replication complexes that were stable in cells containing RNase L suggested that RNase L plays some role, albeit possibly indirect, in the formation of stable replication complexes during alphavirus infection. In addition, confluent monolayers of RNase L-deficient murine cells readily established persistent infections and were not killed. This phenotype is contrary to what has been observed for infection in vertebrate cells with a presumably functional RNase L gene and more resembled alphavirus replication in Aedes mosquito cells, in which the activity of replication complexes making plus stands was also found to decay with inhibition of translation.


Viral and mycoplasmal infections in minor respiratory illnesses and nonstreptococcal pharyngitis. A search for coronaviruses using fetal trachea organ cultures.


[Training of pharmacists].


title: Prevalence and clinical and molecular characterization of human metapneumovirus in children with acute respiratory infection in China.

text: BACKGROUND Human metapneumovirus (HMPV), a newly discovered paramyxovirus, has been associated with acute respiratory tract infections (ARTIs). However, the prevalence and molecular characteristics of HMPV in China are still unclear. METHODS A total of 661 nasopharyngeal aspirates (NPA) specimens were collected from 661 children with ARTIs between December 2006 and November 2008. Specimens were screened for HMPV by reverse transcription-polymerase reaction. All positive amplification products were confirmed by sequencing. RESULTS HMPV was detected in 45 patients (6.80%) of the 661 children. The HMPV-infected patients were from 29 days to 9 years of age. A high incidence of HMPV infection (84.4%) was observed during the winter-spring season. Of the 45 HMPV-positive patients, 25 (55.6%) were co-infected with other respiratory viruses, and respiratory syncytial virus (RSV) was the most common additional respiratory virus. The most common clinical diagnosis was bronchopneumonia (57.8%) and cough (88.9%) was the most common clinical symptom. Phylogenetic analysis of the F gene revealed that 80% of the HMPV detected were A2, 2.2% were A1, and 17.8% were B1. Statistical analyses showed that sex, ages, seasons, and severity of the disease did not correlate with HMPV genotype (P = 0.986, 0.347, 0.660, 0.252), but viral coinfection with HMPV increased hospitalization rates (P = 0.005). CONCLUSIONS HMPV was frequently detected in the pediatric patients with ARTI in China. RSV was the most common coinfection virus and coinfection increased hospitalization rates. All HMPV subgroups except B2 cocirculated and there was no association found between HMPV genotypes and severity of disease.


title: Sequential infection of glial cells by the murine hepatitis virus JHM strain (MHV-4) leads to a characteristic distribution of demyelination.

text: An antigenic variant of the neurotropic murine coronavirus JHMV, designated 2.2-V-1, causes marked demyelination in the relative absence of encephalitis. It is thus useful for the study of the pathogenesis of demyelinating lesions. To better understand the sequential events leading to demyelination, we have examined murine brain and spinal cord tissue at daily intervals after intracerebral inoculation, evaluating them for the distribution of viral antigen, leukocyte infiltration, and demyelination. Immunohistochemical staining indicated that virus established primary infection in the ependymal cells in both brain and spinal cord before spreading into nearby structures and along white matter tracts by cell-to-cell contact. Spread from brain to spinal cord appeared to occur via cerebrospinal fluid. Viral replication was focally cytocidal for ependymal cells, and essentially noncytocidal for other neural cells including glia. In brain, viral antigen and inflammation reached a peak at day 5 postinfection, and rapidly subsided by day 10 postinfection. In spinal cord, viral antigen was less abundant than in brain and was maximal between days 7 and 9 postinfection. The inflammatory response and demyelination, however, were more severe persisting from day 7 through day 19. In the spinal cord, demyelinating lesions developed initially in areas closer to the central canal and were detected most prominently in the anterior funiculi. This finding suggests that the permissiveness of the ependymal cell is crucial to viral entry and that sequential infection of glial cells leads to the characteristic distribution of demyelination.


title: Investigating Turkey Enteric Picornavirus and Its Association with Enteric Disease in Poults.

text: Previous research into the viral community in the poultry gastrointestinal tract has revealed a number of novel and partially described enteric viruses. It is evident that the poultry gut viral community remains minimally characterized and incompletely understood. Investigations into the microbiome of the poultry gut have provided some insight into the geographical distribution and the rapidly evolving taxonomy of the avian enteric picornaviruses. The present investigation was undertaken to produce a comparative metagenomic analysis of the gut virome from a healthy turkey flock versus a flock placed in the field. This investigation revealed a number of enteric picornavirus sequences that were present in the commercial birds in the field that were completely absent in the healthy flock. A novel molecular diagnostic assay was used to track the shedding of field strains of turkey enteric picornavirus in commercial poults inoculated with picornavirus-positive intestinal homogenates prepared from turkeys that were experiencing moderate enteric disease. The propagation of this novel enteric picornavirus in commercial poults resulted in significant reduction in weight gain, and suggests that this common inhabitant of the turkey gut may result in performance problems or enteric disease in the field.


title: Antibiotic resistance in Neisseria gonorrhoeae is diminishing available treatment options for gonorrhea: some possible remedies.

text: Gonorrhea is essentially out of control in many settings and high disease rates are coupled with the spread of multiresistant gonococci. Increases in quinolone resistance have followed loss of the penicillins and tetracyclines as useful treatments. Decreasing susceptibility to third-generation cephalosporins is also reported. Over-reliance on antibiotic treatment as a disease control measure in settings with high disease rates and poor control of antibiotic usage is a significant contributor to the antimicrobial resistance reported. Conversely, containment of resistance is more likely to be achieved when combined with disease control principles shown to be effective. However, until a higher priority is given to funding for sexually transmitted diseases, this prospect is unlikely to eventuate and the possibility of untreatable gonorrhea becomes more real.


title: Lithium chloride inhibits the coronavirus infectious bronchitis virus in cell culture.

text: The avian coronavirus infectious bronchitis virus (IBV) is a major economic pathogen of domestic poultry that, despite vaccination, causes mortality and significant losses in production. During replication of the RNA genome there is a high frequency of mutation and recombination, which has given rise to many strains of IBV and results in the potential for new and emerging strains. Currently the live-attenuated vaccine gives poor cross-strain immunity. Effective antiviral agents may therefore be advantageous in the treatment of IBV. Lithium chloride (LiCl) is a potent inhibitor of the DNA virus herpes simplex virus but not RNA viruses. The effect of LiCl on the replication of IBV was examined in cell culture using two model cell types; Vero cells, an African Green monkey kidney-derived epithelial cell line; and DF-1 cells, an immortalized chicken embryo fibroblast cell line. When treated with a range of LiCl concentrations, IBV RNA and protein levels and viral progeny production were reduced in a dose-dependent manner in both cell types, and the data indicated that inhibition was a cellular rather than a virucidal effect. Host cell protein synthesis still took place in LiCl-treated cells and the level of a standard cellular housekeeping protein remained unchanged, indicating that the effect of LiCl was specifically against IBV.


SARS in health care workers.


BVD virus type 2 infection and severe clinical disease in a dairy herd.


title: High-volume ventilation induces pentraxin 3 expression in multiple acute lung injury models in rats.

text: Pentraxin 3 (PTX3) is an acute-phase protein, which can be produced by a variety of tissue cells at the site of infection or inflammation. It plays an important role in innate immunity in the lung and in mediating acute lung injury. The aim of this study was to determine the effect of mechanical ventilation on PTX3 expression in multiple lung injury models. Male Sprague-Dawley rats were challenged with intravenous injection of lipopolysaccharide (LPS) or hemorrhage followed by resuscitation (HS). The animals were then subjected to either relatively higher (12 ml/kg) or lower (6 ml/kg, positive end-expiratory pressure of 5 cmH(2)O) volume ventilation for 4 h. High-volume ventilation significantly enhanced PTX3 expression in the lung, either alone or in combination with LPS or hemorrhage. A significant increase of PTX3 immunohistochemistry staining in the lung was seen in all injury groups. The PTX3 expression was highly correlated with the severity of lung injury determined by blood gas, lung elastance, and wet-to-dry ratio. To determine the effects of HS, LPS, or injurious ventilation (25 ml/kg) alone on PTX3 expression, another group of rats was studied. Injurious ventilation significantly damaged the lung and increased PTX3 expression. A local expression of PTX3 induced by high-volume ventilation, either alone or in combination with other pathological conditions, suggests that it may be an important mediator in ventilator-induced lung injury.


title: Duration of immunity in cats after vaccination or naturally acquired infection.

text: The necessity for cats to be vaccinated annually against common pathogens has been questioned because sarcomas have infrequently been reported at the injection site. However, with few exceptions, the duration of immunity induced by vaccination or infection is uncertain, and there may therefore be a risk associated with a decision not to revaccinate. This article reviews the information available about the duration of immunity induced by vaccination or infection in cats, and reveals many shortcomings that make blanket recommendations impossible. Each vaccine must be considered individually.


title: Evaluation of Australia's National Notifiable Disease Surveillance System.

text: The Australian National Notifiable Diseases Surveillance System (NNDSS) is a passive surveillance system that collects information on communicable diseases. The Australian Government manages NNDSS under the auspices of the Communicable Diseases Network Australia (CDNA). Data collected by each state and territory are collated, analysed and disseminated by the Australian Government Department of Health and Ageing. We report the first evaluation of NNDSS since it was established in 1991. Three primary stakeholder groups were surveyed: (a) CDNA members, (b) the National Surveillance Committee and (c) the readership of Communicable Diseases Intelligence, the primary means of data dissemination from NNDSS. The evaluation revealed that the system was acceptable, structurally simple, and that the data collected were actively used by stakeholders. However, the lack of clearly documented aims and objectives for NNDSS, inflexibility to changing needs, lack of timeliness and complexity in processes were seen as problematic. The results of this evaluation, supported by recent federal funding to enhance national biosecurity, will provide the framework for enhancing NNDSS to meet national communicable disease surveillance requirements in Australia.


title: A consensus statement on the use of angiotensin receptor blockers and angiotensin converting enzyme inhibitors in relation to COVID-19 (corona virus disease 2019).

text: There has been a lot of speculation that patients with coronavirus disease 2019 (COVID-19) who are receiving angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor blockers (ARBs) may be at increased risk for adverse outcomes. We reviewed the available evidence, and have not found this to be the case. We recommend that patients on such medications should continue on them unless there is a clinical indication to stop their use.