title: Soluble receptor-mediated targeting of mouse hepatitis coronavirus to the human epidermal growth factor receptor.

text: The mouse hepatitis coronavirus (MHV) infects murine cells by binding of its spike (S) protein to murine CEACAM1a. The N-terminal part of this cellular receptor (soR) is sufficient for S binding and for subsequent induction of the conformational changes required for virus-cell membrane fusion. Here we analyzed whether these characteristics can be used to redirect MHV to human cancer cells. To this end, the soR domain was coupled to single-chain monoclonal antibody 425, which is directed against the human epidermal growth factor receptor (EGFR), resulting in a bispecific adapter protein (soR-425). The soR and soR-425 proteins, both produced with the vaccinia virus system, were able to neutralize MHV infection of murine LR7 cells. However, only soR-425 was able to target MHV to human EGFR-expressing cancer cells. Interestingly, the targeted infections induced syncytium formation. Furthermore, the soR-425-mediated infections were blocked by heptad repeat-mimicking peptides, indicating that virus entry requires the regular S protein fusion process. We conclude that the specific spike-binding property of the CEACAM1a N-terminal fragment can be exploited to direct the virus to selected cells by linking it to a moiety able to bind a receptor on those cells. This approach might be useful in the development of tumor-targeted coronaviruses.


title: Central nervous system infections - a potential complication of systemic immunotherapy.

text: PURPOSE OF REVIEW Multiple sclerosis is a chronic inflammatory disease of the central nervous system, characterized by demyelination and axonal damage. With the emergence of more efficient immune therapies, severe sometimes even fatal central nervous system infections were observed. This review will address the role of the systemic immune system in central nervous system immune surveillance and discuss implications for the development of novel immunotherapies in multiple sclerosis. RECENT FINDINGS In the last decade, a number of immunosuppressive and immunomodulatory agents have been approved for the treatment of multiple sclerosis, based on the results of double-blind placebo-controlled randomized clinical trials. A better understanding of the pathogenesis of the disease has prompted exploration of the therapeutic utility of new drugs that more potently disrupt the pathophysiological cascade of events that culminates in central nervous system tissue damage. One potential side-effect of these newer therapies is their interference with the control of central nervous system infections by the immune system. SUMMARY Any increase in potency of multiple sclerosis therapies could well interfere with protective immunosurveillance of the central nervous system. One possible outcome is an increased incidence of opportunistic infections. A heightened vigilance for central nervous system infections in the setting of immunosuppression is necessary to prevent serious side-effects in the future.


title: Antigen-capture blocking enzyme-linked immunosorbent assay based on a baculovirus recombinant antigen to differentiate Transmissible gastroenteritis virus from Porcine respiratory coronavirus antibodies.

text: A new commercially available antigen-capture, blocking enzyme-linked immunosorbent assay (antigen-capture b-ELISA), based on baculovirus truncated-S recombinant protein of Transmissible gastroenteritis virus (TGEV) and 3 specific monoclonal antibodies, was developed and evaluated by examining a panel of 453 positive Porcine respiratory coronavirus (PRCoV), 31 positive TGEV, and 126 negative field sera by using another commercially available differential coronavirus b-ELISA as the reference technique to differentiate TGEV- from PRCoV-induced antibodies. The recombinant S protein-based ELISA appeared to be 100% sensitive for TGEV and PRCoV detection and highly specific for TGEV and PRCoV detection (100% and 92.06%, respectively), when qualitative results (positive or negative) were compared with those of the reference technique. In variability experiments, the ELISA gave consistent results when the same serum was evaluated on different wells and different plates. These results indicated that truncated recombinant S protein is a suitable alternative to the complete virus as antigen in ELISA assays. The use of recombinant S protein as antigen offers great advantages because it is an easy-to-produce, easy-to-standardize, noninfectious antigen that does not require further purification or concentration. Those advantages represent an important improvement for antigen preparation, in comparison with other assays in which an inactivated virus from mammalian cell cultures is used.


title: pH stability studies with avian infectious bronchitis virus (coronavirus) strains.

text: A comparison of 17 infectious bronchitis virus strains, using the same test procedure and assay system, demonstrated that stability at an acid pH is a variable characteristic of the avian coronaviruses.


title: Parasites from Indo-Pacific hump-backed dolphins (Sousa chinensis) and finless porpoises (Neophocaena phocaenoides) stranded in Hong Kong.

text: Between 1993 and 1998, 28 Indo-Pacific hump-backed dolphins (Sousa chinensis) and 32 finless porpoises (Neophocaena phocoenoides) stranded in Hong Kong territorial waters were examined postmortem for parasites. The nematode Halocercus pingi was discovered in the lungs of one hump-backed dolphin and in 10 finless porpoises, typically within abscesses or granulomata, and they were frequently accompanied by a catarrhal exudate and lesions characteristic of pneumonia. Seven of the 10 finless porpoises were calves with substantial lungworm infections, and three were neonates with visible fetal folds and umbilical remnants, suggesting that H pingi is transferred to the neonate before birth or during lactation. Electron micrographs of H pingi should allow the nematode to be identified by other researchers. An ectoparasitic stalked barnacle (Xenobalanus globicipitis) was recovered from a finless porpoise, the first time that this species of barnacle has been recorded in Hong Kong's territorial waters.


title: Multiplex fluorescent analysis of four short tandem repeats for rapid haemophilia A molecular diagnosis.

text: Indirect molecular diagnosis of hemophiliaA (HA) is carried out by analyzing intragenic polymorphic markers described along the coagulation factorVIII (FVIII) gene. Several studies have demonstrated that the two commonly used intronic short tandem repeats (STR13 and STR22) located in the FVIII gene are highly informative for this task. Two extragenic markers closely linked to FVIII (DXS1073 and DXS1108) have also been described as valuable tools for gene tracking. The objective of the present work was to develop a rapid, single-tube automated method to simultaneously analyze these four STRs. Consistent amplification was achieved by quadruplex fluorescent PCR and the products were analyzed by capillary electrophoresis. Validation of the method included DNA analysis of 88 individuals from a control population, 45 HA patients and 32 individuals from 10 HA-affected families. Statistical study showed that the STR13, STR22 and DXS1108 loci were in significant linkage disequilibrium, whereas DXS1073 was not. Nevertheless, the combination of the four markers offered a high heterozygosity rate (>90%) that improved tracing of FVIII gene inheritance. Optimal results with application to single cells in a HA preimplantation genetic diagnosis (PGD) protocol demonstrated the sensitivity of the technique. In conclusion, the automated fluorescent method described is an extremely rapid, simple and highly informative one that is easy to standardize and allows direct comparison of results among different groups working with genetic counseling, prenatal diagnosis and PGD in HA-affected families.


title: Induction of protective immunity against transmissible gastroenteritis virus after exposure of neonatal pigs to porcine respiratory coronavirus.

text: OBJECTIVE To test the ability of porcine respiratory coronavirus (PRCV) to induce protective immunity to antigenically related transmissible gastroenteritis virus (TGEV) in neonatal pigs. DESIGN Neonatal pigs were exposed to PRCV when they were 2, 4, or 6 days old and challenge-exposed to virulent TGEV at 10 days of age. ANIMALS 34 hysterectomy-derived, colostrum-deprived pigs. PROCEDURE After challenge exposure, clinical signs were observed, body weight, antibody response, and virus shedding were measured, and mortality was determined. RESULTS After exposure to PRCV, principals had a slightly slower rate of weight gain than did controls; with 1 exception (a PRCV-exposed pig that was dyspneic for 1 day), principals and controls remained clinically normal until shortly after challenge exposure, when all pigs became listless and anorectic and developed watery diarrhea. However, by day 3, most of the pigs that had been exposed to PRCV when they were either 2 or 4 days old began to recover and most (15/18) survived. Conversely, the clinical condition of most of the other pigs worsened and most (14/16) died. Pigs exposed to PRCV when they were 2 or 4 days old also differed from all other pigs in that they had serum virus-neutralizing antibodies for PRCV and TGEV at the time of challenge exposure. CONCLUSIONS The PRCV can induce protective immunity to TGEV in neonatal pigs and such immunity develops at or about 6 days after exposure to PRCV. Moreover, protective immunity may be coincident with the appearance of virus-neutralizing antibody. CLINICAL RELEVANCE Exposure to PRCV should enhance a TGE herd vaccination program.


title: Early experience with elective single-port laparoscopic appendectomy and cholecystectomy in day-case surgery.

text: BACKGROUND Single incision laparoscopy aims to reduce access trauma and facilitate specimen extraction. We report our early experience and describe the learning curve for single-port laparoscopic appendectomy and cholecystectomy as a primer for more advanced single-port laparoscopic procedures. METHODS From June 2009 to February 2010, all consecutive patients undergoing elective single-port appendectomy and cholecystectomy at the day surgical centre were included. A prospective database with perioperative data was created. RESULTS Ten appendectomies and 4 cholecystectomies were done in 13 patients, predominantly males (77%), with a mean age of 45 years and a mean BMI of 23 kg/m2. The mean operative time was 31 minutes and 75 minutes for appendectomy and cholecystectomy, respectively. One extra trocar was placed in the first two cholecystectomies, to improve exposure and allow cholangiography. All patients did well at a median follow-up of 5.8 months and no wound infections, nor incisional hernias were detected. CONCLUSION Single-port laparoscopic appendectomy and cholecystectomy is safe and feasible with a short learning curve and has good outcome.


title: You Need a Plan: A Stepwise Protocol for Operating Room Preparedness During an Infectious Pandemic.

text: Background The worldwide spread of SARS-CoV-2, the coronavirus that causes the syndrome designated COVID-19, presents a challenge for emergency operative management. The transmission and virulence of this new pathogen has raised concern for how best to protect operating room staff while effectively providing care to the infected patient requiring urgent or emergent surgery. Observations Establishment of a clear protocol that adheres to rigorous infection control measures while providing a safe system for interfacility transport and operative care is vital to a successful surgical pandemic response. While emergency protocols must be rapidly developed, they should be collaboratively improved and incorporate new knowledge as and when it becomes available. These measures combined with practice drills to keep operating room personnel ready and able should help construct processes that are useful, easy to follow, and tailored to the unique local environment of each health care setting. Conclusions After the initial apprehensions and struggles during our confrontation with the COVID-19 crisis, it is our hope that the experience we share will be helpful to surgical staff at other institutions grappling with the challenges of operative care in the pandemic environment. While this protocol focuses on the current COVID-19 pandemic, these recommendations serve as a template for surgical preparedness that can be readily adapted to infectious disease crisis that unfortunately might emerge in the future.


title: Genetic recombination during coinfection of two mutants of human respiratory syncytial virus.

text: Recombination between coinfecting viruses had not been documented previously for a nonsegmented negative-strand RNA virus (mononegavirus). We investigated the potential of intermolecular recombination by respiratory syncytial virus (RSV) by coinfecting HEp-2 cells with two recombinant RSV (rRSV) mutants lacking either the G gene (DeltaG/HEK) or the NS1 and NS2 genes (DeltaNS1/2). These viruses replicate inefficiently and form pinpoint plaques in HEp-2 cells. Therefore, potential recombined viruses with a growth and/or plaque formation advantage should easily be identified and differentiated from the two parental viruses. Further identification of potential recombinants was aided by the inclusion of point mutation markers in the F and L genes of DeltaG/HEK and the design of reverse transcription-PCR (RT-PCR) primers capable of detecting these markers. Independent coinfections and control single infections by these two rRSV mutants were performed. In one of six coinfections, an RSV variant was identified that produced plaques slightly larger than those of wild-type RSV in HEp-2 cells. RT-PCR and sequencing provided evidence that this variant was a recombined RSV (rec-RSV). The rec-RSV appeared to have been generated by a polymerase jump from the DeltaG/HEK genome to that of DeltaNS1/2 and back again in the vicinity of the SH-G-F genes. This apparently involved nonhomologous and homologous recombination events, respectively. The recombined genome was identical to that of the DeltaG/HEK mutant except that all but the first 12 nucleotides of the SH gene were deleted and replaced by an insert consisting of the last 91 nucleotides of the G gene and its downstream intergenic region. This insert could have come only from the coinfecting DeltaNS1/2 virus. This resulted in the formation of a short chimeric SH:G gene. Northern and Western blot analysis confirmed that the rec-RSV did not express the normal SH and G mRNAs and proteins but did express the aberrant SH:G mRNA. This provides an experimental demonstration of intermolecular recombination yielding a viable, helper-independent mononegavirus. However, the isolation of only a single rec-RSV under these optimized conditions supports the idea that RSV recombination is rare indeed.


title: An enteric coronavirus infection of cats and its relationship to feline infectious peritonitis.

text: An enteric coronavirus that is antigenically closely related to feline infectious peritonitis virus (FIPV) is ubiquitous in the cat population. This virus has been designated feline enteric coronavirus to differentiate it from FIPV. The virus is shed in the feces by many seropositive cats; in catteries it is a cause of inapparent to mildly severe enteritis in kittens 6 to 12 weeks of age. The virus may produce a more severe enteritis in young specific-pathogen-free kittens. Feline enteric coronavirus selectively infects the apical columnar epithelium of the intestinal villi, from the caudal part of the duodenum to the cecum. In severe infections, there are sloughing of the tips of the villi and villous atrophy. Many cats recovering from the disease remain carriers of the virus. Recovered cats, observed for 3 to 24 months, remained healthy and did not develop peritonitis, pleuritis, or granulomatous disease. The relationship of feline enteric coronavirus and FIPV was studied. Although the viruses were antigenically similar, they were distinctly different in their pathogenicities. The enteric coronavirus did not cause feline infectious peritonitis in coronavirus antibody-negative cats inoculated orally or intraperitoneally nor in coronavirus antibody-positive cats inoculated intraperitoneally or intratracheally. Serologic tests, using FIPV, canine coronavirus, and transmissible gastroenteritis virus of swine as substrate antigens in fluorescent antibody procedures may not accurately identify FIPV infection. These tests do not appear to distinguish between FIPV and this feline enteric coronavirus.


title: Evaluation and management of illness in a child after international travel.

text: International travel by children is becoming increasingly common over the last few decades. As a result, clinicians may be asked to assist in the evaluation and treatment of an ill child after international travel. Along with recording the typical thorough history of present illness and medical history, the physician who cares for a child in this situation will have to take a thorough travel history and consider not only local conditions, but also possible pathogens that the child may have encountered during the journey. A detailed travel history should be routinely performed to investigate specific exposures. Reference materials are available that can guide the clinician in determining which infectious agents are endemic to various areas of the world. With the exception of obtaining a basic laboratory screen, further evaluation of the young patient should be dictated by the most likely cause of the illness. Repeated testing and examinations and close follow-up may be necessary to arrive at the proper diagnosis and initiate appropriate treatment in a timely manner.


title: Antigen presentation by astrocytes primes rat T lymphocytes for apoptotic cell death. A model for T-cell apoptosis in vivo.

text: In this study we have characterized apoptotic cell death of autoreactive T cells resulting from their interaction with astrocytes and the modulatory effect of steroid hormones. Time kinetics of T-cell activation by interferon (IFN)-gamma-treated astrocytes from neonatal Lewis rats and by professional antigen presenting cells (APCs) from bulk suspensions of thymus or spleen were performed. [3H]Thymidine incorporation of neuritogenic P2- and encephalitogenic myelin basic protein (MBP)-specific T-cell lines declined after 48 h in culture with astrocytes. A similar suppressive effect was observed when T cells were cocultured with thymic APCs and astrocytes. This effect disappeared when astrocytes were separated by a transwell system. After 72 h of culture with astrocytes a mean of 17.5 +/- 12.4% T cells exhibited morphological signs of apoptosis. Apoptosis was identified by light microscopy, and confirmed by electron microscopy, by in situ tailing reaction and by agarose gel electrophoresis. Glucocorticosteroids and oestrogen specifically enhanced T-cell apoptosis within 8 h (69.8 +/- 23.1% and 69 +/- 17.1%, respectively) when added after 72 h to the astrocyte system, but not at earlier time points of T-cell activation or when thymic APCs were used. Glucocorticoid-mediated T-cell apoptosis was inhibited by the steroid-receptor antagonist RU 486. Pregnenolone, lipocortin-1, indomethacin and transforming growth factor-beta did not induce apoptosis in this system. The steroid effect was not associated with CD28, IL-2 receptor, or transferrin-receptor expression, which were equally upregulated on T cells activated by astrocytes or thymic APC as shown by fluorescence activated cell sorting (FACS) analysis. We conclude that astrocytes as CNS-specific APC may render T cells susceptible for induction of apoptotic cell death. Some steroid hormones can markedly enhance this process in vitro and may augment an additional effect of a systemic corticosteroid response in vivo during recovery from autoimmune encephalomyelitis.


title: [SARS. Severe acute respiratory syndrome].

text: Since November 2002 an epidemic of a new severe respiratory infection has taken place primarily in South-East Asia. Severe Acute Respiratory Syndrome (SARS) is assumed caused by a recently identified corona virus, which seems to be a new pathogen in the human population. The infection is assumed transferred primarily with droplets and is highly contagious within healthcare settings and families. Respiratory and barrier precautions are necessary to contain the virus, as it is contact tracing.


title: Advanced glycation end products stimulate tumor necrosis factor-alpha and interleukin-1 beta secretion by peritoneal macrophages in patients on continuous ambulatory peritoneal dialysis.

text: BACKGROUND Advanced glycation end products, formed by the non-enzymatic glycation of proteins with reducing sugars, are thought to play a pathogenetic role in the vascular complications of diabetes, uremia and atherosclerosis. 132-microglobulin is a major constituent of amyloid fibrils in dialysis-related amyloidosis. AGE1-modified beta2m has been found in amyloid deposits of long-term hemodialysis patients. AGE-modified 132m has also been shown to enhance chemotaxis and increase tumor necrosis factor-alpha and interleukin-1 beta secretion by circulating and tissue monocytes/macrophages. OBJECTIVES To investigate the effect of AGE-modified 132m and AGE-human serum albumin on TNF-alpha and IL-1beta secretion by human peritoneal macrophages derived from patients on continuous ambulatory peritoneal dialysis. METHODS Human PMO were isolated from peritoneal dialysis effluent of stable CAPD patients and were incubated for 24 hours with AGE-modified beta2m, beta2m, AGE-HSA, HSA or lipopolysaccharide. TNF-alpha or IL-1beta secretion was measured by enzyme-linked immunosorbent assay in cell-free culture supernatants. RESULTS Both AGE-modified beta2m and AGE-HSA significantly increased TNF-alpha and IL-1beta secretion by human PMO in a dose-dependent manner (50-200 microg/ml). In contrast, beta2m or HSA had no such stimulatory effect on TNF-alpha secretion but had a small significant increase in IL-1beta secretion. CONCLUSIONS AGE-modified beta2m promotes in vitro TNF-alpha and IL-13 secretion by human PMO of CAPD patients. Activation of these macrophages by AGE-modified beta2m may be a contributory factor to the morphologic changes and altered permeability of the peritoneal membrane in long-term CAPD.


title: Detection of bovine viral diarrhea virus by spot hybridization with probes prepared from cloned cDNA sequences.

text: A cytopathic strain of bovine viral diarrhea virus (BVDV) was purified from infected cell culture fluids by isopycnic density-gradient centrifugation. Genomic RNA was extracted and tailed with adenine residues at the 3' end with poly-A polymerase. Double-stranded complementary DNA (cDNA) was synthesized, using the poly-A-tailed RNA as a template and oligo-dT as a primer, and then cloned into the pUC9 plasmid. Virus-specific cDNA sequences, varying in length from 0.5 to 2.5 kilobases (kb), were obtained. One BVDV-specific sequence of cloned cDNA, 1.1 kb in length and with an internal Pst I restriction endonuclease cleavage site, was selected for use as a probe. The cloned cDNA insert was removed from the plasmid either with or without flanking plasmid sequences and labeled with 32P-nucleotides by nick translation for use as hybridization probes for BVDV. The performance of probes of smaller fragments of the insert was compared to that of the intact sequence in hybridization assays. In addition, 2 methods of specimen preparation were compared to establish optimum parameters for hybridization. The hybridization assay was 10-100 times more sensitive than infectivity assays for BVDV in infected cell cultures. Freezing of specimens reduced by 10-fold the sensitivity of hybridization for BVDV target sequences. The probes prepared from the cloned cDNA hybridized with all cytopathic and noncytopathic BVDV strains tested but not with uninfected cell cultures, cellular ribosomal RNA, bovine coronavirus, bluetongue virus, or bovine adenovirus 3. Probes prepared with native plasmid DNA did not hybridize with BVDV or uninfected cell cultures.(ABSTRACT TRUNCATED AT 250 WORDS)


title: Single Incision Pediatric Endoscopic Surgery: Advantages of a Relatively Large Incision.

text: OBJECTIVE To describe Single Incision Pediatric Endoscopic Surgery (SIPES) performed on children with various diagnoses, emphasizing its advantages. STUDY DESIGN An observational case series. PLACE AND DURATION OF STUDY Department of Pediatric Surgery, Dr. Sami Ulus Maternity and Child Health Hospital, Ankara, Turkey, from January 2011 to November 2014. METHODOLOGY A review of patient charts was conducted in which SIPES was preferred as the surgical procedure. Patient demographics, operative details, operative time, clinical outcomes, postoperative pain and cosmesis were analyzed. RESULTS SIPES was performed on 45 patients (21 girls, 24 boys). Thirty-three appendectomies, 5 varicocelectomies, 3 oophorectomies, 2 ovarian and one paratubal cyst excision, and one fallopian tube excision were performed. All except one procedures were performed through our standard 2 cm umbilical vertical or smile incision. In 18 cases, abdominal irrigation/aspiration was easily performed through the existing larger incision, as is done with open surgical technique. None of the patients had early postoperative shoulder/back pain since complete disinflation of CO2could be ensured. All of the patients/parents were satisfied with the cosmesis. CONCLUSION SIPES has the advantages of limiting the surgical scar to within the umbilicus and providing easy disinflation of CO2, allowing intraabdominal cleaning and extraction of large volume tissue samples through a single large umbilical incision.


title: The new coronavirus that came from the East: analysis of the initial epidemic in Mexico.

text: INTRODUCTION As of March 23, 2020, suspension of non-essential activities was declared in Mexico throughout the country in order to mitigate the spread of the COVID-19 pandemic. OBJECTIVE To analyze data on the first 1,510 laboratory-confirmed cases of COVID-19 in Mexico, and to describe the geographical distribution of the disease and its transmission dynamics. METHOD Description of the first COVID-19 cases with real-time RT-PCR-positive test, as well as evaluation of epidemiological measures, cumulative incidence, rate of transmission, and mortality and lethality rates during the first month of the epidemic. RESULTS Average age was 43 years, and 58 % were males; 44 % of initial cases were imported. Lethality in the population during the first month went from 1.08 to 3.97 per 100 cases; however, the trend is linear and similar to that observed in Europe. CONCLUSIONS In Mexico, social distancing is being applied, but studies are still required on the dynamics of the epidemic, person-to-person transmission, incidence of subclinical infections, and patient survival.


title: Frequency of interferon-alpha-secreting blood leukocytes in irradiated and bone-marrow-grafted pigs.

text: The effects of irradiation were studied on porcine interferon-alpha (IFN-alpha) secreting cells (IFN-alpha SC). IFN-alpha SC were characterized by an ELISPOT assay on non-adherent PBMC following incubation with the transmissible gastroenteritis coronavirus. In vitro irradiation of PBMC was followed by a decrease in the number of IFN-alpha SC while IFN-gamma production and cell viability were not affected. These data indicate that porcine IFN-alpha SC are relatively radiosensitive. Indeed, the frequency of blood IFN-alpha SC decreased markedly and rapidly after in vivo whole body or partial lymphoid irradiation. In addition, within several days of compatible bone-marrow engraftment in the irradiated animals, the number of blood IFN-alpha SC returned to normal values. These data demonstrate that circulating porcine IFN-alpha SC are derived from bone-marrow progenitors.


title: Parasite prevalence in free-ranging farm cats, Felis silvestris catus.

text: No animals tested were positive for feline leukemia virus antigen and Chlamydia psittaci antibodies, but all were positive for antibodies to feline calicivirus (FCV), feline herpesvirus 1 (FHV1) and rotavirus. They had antibodies to feline parvovirus (96%), feline coronavirus (84% and cowpox virus (2%). Antibody to feline immunodeficiency virus (FIV) was found in 53% of animals, which were less likely to be infected with Haemobartonella felis, and had higher FHV antibody titres than cats without FIV. FCV was isolated from 51% cats and FHV1 and feline reovirus each from 4%. H. felis was present in 42% of animals, and antibody to Toxoplasma gondii in 62%. Clinical abnormality had a significant association with FIV and feline calicivirus infections, but sex, age, social status and feeding group had no significant association with prevalence of any parasites. Toxocara cati and Toxascaris leonina eggs were found, respectively, in 91% and 82% of animals tested.


title: Discontinuous subgenomic RNA synthesis in arteriviruses is guided by an RNA hairpin structure located in the genomic leader region.

text: Nidoviruses produce an extensive 3'-coterminal nested set of subgenomic (sg) mRNAs, which are used to express structural proteins and sometimes accessory proteins. In arteriviruses and coronaviruses, these mRNAs contain a common 5' leader sequence, derived from the genomic 5' end. The joining of the leader sequence to different segments derived from the 3'-proximal part of the genome (mRNA bodies) presumably involves a unique mechanism of discontinuous minus-strand RNA synthesis in which base pairing between sense and antisense transcription-regulating sequences (TRSs) plays an essential role. The leader TRS is present in the loop of a hairpin structure that functions in sg mRNA synthesis. In this study, the minimal sequences in the 5'-proximal region of the Equine arteritis virus genome that are required for sg RNA synthesis were delimited through mutagenesis. A full-length cDNA clone was engineered in which this domain was duplicated, allowing us to make mutations and monitor their effects on sg RNA synthesis without seriously affecting genome replication and translation. The leader TRS present in the duplicated sequence was used and yielded novel sg mRNAs with significantly extended leaders. Our combined findings suggest that the leader TRS hairpin (LTH) and its immediate flanking sequences are essential for efficient sg RNA synthesis and form an independent functional entity that could be moved 300 nucleotides downstream of its original position in the genome. We hypothesize that a conformational switch in the LTH region regulates the role of the 5'-proximal region of the arterivirus genome in subgenomic RNA synthesis.


title: [Radiological manifestation of COVID-19].

text: In this case report, a 70-year-old male was admitted to the hospital with respiratory tract symptoms and a fever, and he was isolated, suspected of COVID-19. A chest radiograph showed bilateral consolidations, and subsequent chest CT scan revealed bilateral and peripheral ground glass opacities, which are the most common radiological manifestations of COVID-19. The characteristic radiological findings can be exploited in the diagnosis of COVID-19, and radiologists should have knowledge of the usual manifestations of COVID-19, as they may play a role in early detection and diagnosis of the disease.


title: Clinical Trial of an Educational Program to Decrease Monitor Alarms in a Medical Intensive Care Unit.

text: Clinical research to identify effective interventions for decreasing nonactionable alarms has been limited. The objective of this study was to determine if a staff educational program on customizing alarm settings on bedside monitors decreased alarms in a medical intensive care unit (MICU). A preintervention, postintervention, nonequivalent group design was used to evaluate an educational program on alarm management in a convenience sample of MICU nurses. A 15-minute session was provided in a 1-week period. The outcome variable (number of alarms for low oxygen saturation via pulse oximetry [SpO2]) was determined from monitor log files adjusted by patient census. Data were collected for 15 days before and after the intervention. χ2 analysis was used, with P less than .05 considered significant. After 1 week of education, low SpO2 alarms decreased from 502 to 306 alarms per patient monitored per day, a 39% reduction (P < .001). Instructions for nurses in the medical intensive care unit on individualizing alarm settings to patients' clinical condition decreased common monitor alarms by 39%.


title: Preclinical evaluation of two real-time, reverse transcription-PCR assays for detection of the severe acute respiratory syndrome coronavirus.

text: We verified the analytical performance characteristics of a previously described real-time reverse transcription-PCR (RT-PCR) assay targeting the open reading frame (ORF) 1b region of the severe acute respiratory syndrome coronavirus (SARS-CoV) with RNA transcripts. We then compared it to a novel nucleocapsid gene real-time RT-PCR assay with genomic RNA. The assays differed only in the primer and probe sequences and final concentrations. A commercially available armored RNA (Ambion, Austin, Tex.) was evaluated as positive control for the ORF 1b assay. The analytical sensitivity, reproducibility, amplification efficiency, and dynamic range of the assays were similar. Both were specific for SARS-CoV as determined by testing against human CoV 229E and OC43, specimens from patients without SARS, and by BLAST searches of GenBank for primer and probe sequence homology. The armored RNA was found to be a suitable positive control for the ORF 1b assay that could be reliably recovered and amplified from a variety of clinical specimens.


The making of infectious viral RNA: No size limit in sight.


title: Drug use trends for arthritis and other rheumatic conditions and effect of patient's age on treatment choice.

text: BACKGROUND Little is known about the influence of the patient's age on selection of treatment for arthritis and other rheumatic conditions (AORCs). The aim of the present study was to examine drug use trends in patients with AORCs in ambulatory care and to study age-specific drug use. METHODS Data from the National Ambulatory Medical Care Survey and the outpatient department component of the National Hospital Ambulatory Medical Care Survey were used to examine ambulatory visits for AORCs during 2001-2005. Use of nonsteroidal anti-inflammatory drugs (NSAIDs), analgesics (narcotics and nonnarcotics), steroids, and disease-modifying antirheumatoid drugs (DMARDs) was measured as the proportion of visits that resulted in a prescription. Age-specific drug use trends were determined separately for adult (19-64 years) and for elderly (> or = 65 years) patients. RESULTS According to the national surveys, there were 208 million ambulatory visits for AORCs during the study period. Use of NSAIDs, analgesics, steroids, and DMARDs was found to be 33.3%, 23.5%, 15.7%, and 4.3%, respectively. Analgesic use increased from 18.3% in 2001 to 26.7% in 2005. DMARDs and NSAIDs were prescribed 1.2 times more to adult patients than to elderly patients. Steroids and analgesics were prescribed 1.3 and 1.2 times more, respectively, to elderly patients than to adult patients. LIMITATION The findings do not translate to population-based prevalence measures, as the unit of analysis was the patient visit. CONCLUSIONS NSAIDs remain the most frequently prescribed drug class for AORCs; however, there has been increased use of analgesics in recent years. The study findings suggest that the patient's age plays a role in the choice of drugs prescribed for AORCs.


title: A brief review of infectious and parasitic diseases of wapiti, with emphasis on western Canada and the northwestern United States.

text: In this paper I review diseases reported in both captive and free-ranging wapiti in western North America, with some reference to diseases in captive red deer in Great Britain, Europe, New Zealand, and eastern North America.With the exception of coronavirus in neonates, few viral agents are reported to cause serious disease losses in wapiti in North America at this time. Bacterial diseases of current significance include brucellosis (focus in Wyoming), clostridial diseases, coliform enteritis of neonates, pasteurellosis, and necrobacillosis. The endoparasites most likely to be seen causing lesions in wapiti of western North America are lungworm (Dictyocaulus viviparus), arterial worm (Elaeophora schneideri), and, possibly, liver fluke (Fascioloides magna). Ectoparasites of importance to wapiti are Psoroptes cervinus and Dermacentor albipictus. Nutritional diseases are not covered in this review.


title: Species-specific exclusion of APOBEC3G from HIV-1 virions by Vif.

text: The HIV-1 accessory protein Vif (virion infectivity factor) is required for the production of infectious virions by CD4(+) lymphocytes. Vif facilitates particle infectivity by blocking the inhibitory activity of APOBEC3G (CEM15), a virion-encapsidated cellular protein that deaminates minus-strand reverse transcript cytosines to uracils. We report that HIV-1 Vif forms a complex with human APOBEC3G that prevents its virion encapsidation. HIV-1 Vif did not efficiently form a complex with mouse APOBEC3G. Vif dramatically reduced the amount of human APOBEC3G encapsidated in HIV-1 virions but did not prevent encapsidation of mouse or AGM APOBEC3G. As a result, these enzymes are potent inhibitors of wild-type HIV-1 replication. The species-specificity of this interaction may play a role in restricting HIV-1 infection to humans. Together these findings suggest that therapeutic intervention that either induced APOBEC3G or blocked its interaction with Vif could be clinically beneficial.


title: A pilot study on the effects of curd (dahi) & leaf protein concentrate in children with protein energy malnutrition (PEM).

text: BACKGROUND & OBJECTIVE In protein-energy malnutrition (PEM) there is a significant impairment of immunity, both cell-mediated and humoral, which may be reversed with nutritional rehabilitation. With the use of probiotics like curd (dahi) and micronutrient-rich leaf protein concentrate (LPC), this immune recovery may be hastened. This study was conducted to assess the impact of supplementation of curd and LPC on nutritional status, and immunity as assessed by anthropometry, haemoglobin, ferritin levels, T- cell subpopulation and C-reactive protein (CRP), in children suffering from PEM. METHODS Eighty moderate to severely malnourished children (1-5 yr) were randomized to receive either curd or LPC in addition to WHO recommended two-step diet over 15 days. Nutritional, immunological and haematological parameters were measured before and after supplementation and compared within the groups. RESULTS The change in weight, haemoglobin level and CD4:CD8 T-cell subpopulation was significant in both the groups after supplementation. Response of CRP was blunted in PEM. Serum ferritin decreased significantly after supplementation in both groups. INTERPRETATION & CONCLUSION Curd and LPC when added to diet of malnourished children, may have therapeutic value by accelerating immune recovery. More studies need to be done on a larger sample to confirm these findings.


title: [Cloning, purification, and antigenic characterization of three recombinant fragments derived from SARS-CoV S1 domain].

text: OBJECTIVE The present study aimed to clone and express three fragments of genomic RNA derived from SARS associated coronavirus (SARS-CoV) S1 domain and to study its immunogenicity. METHODS The S1 domain gene was amplified by PCR with specific primers and was inserted into the prokaryotic expression vector pQE-30. Three fragments (40-751, 746-1344 and 746-2001 bp) derived from S1 domain produced after the recombinant plasmid (pQE-30/S1) was digested by restriction endonucleases. The three fragments were cloned into pQE-30 and expressed in M15 strains of Escherichia coli. The expression products, designated S1a, S1b and S1c respectively, were purified by Ni affinity chromatography. The immunogenicity was analyzed by Western Blot and ELISA using serologically confirmed sera from SARS patients and the sera from healthy donors was used as control at the same assay. RESULTS Three recombinant plasmids (pQE-30/S1a, pQE-30/S1b, pQE-30/S1c) were constructed.Fusion proteins with relative molecular mass of 26,700, 22,500 and 46,000 dalton were successfully expressed with amounts of 35%, 35% and 30% of total cell protein and purified by Ni affinity chromatography, respectively. Western Blot and ELISA analysis showed that the S1c protein could be specifically recognized by the sera from SARS patients. CONCLUSION The recombinant S1c protein was a good immunogen and has the potential to be used as a vaccine against SARS-CoV infection.


Molecular biology and pathogenesis of coronaviruses.


title: [The response of intensive care medicine in situations of multiple victims and catastrophe].

text: There is a change in attitude regarding critical patients in situations of multiple victims and disasters in order to give the better treatment to the largest number of patients. Recent large-scale accidents, including terrorist attacks and biological warfare as the outbreak of Severe Acute Respiratory Syndrome (SARS) in 2002/2003 and pandemic influenza have demonstrated the need for an adequate response from Intensive Care Medicine in those contexts. It is essential, according to risk analysis, organizing and planning the response of the Intensive Care Units in accordance with its strategy to increase the capacity of beds in intensive care. It is necessary a permanent preparation of the Intensive Care Units to face exceptional situations, giving focus to the training of professionals, triage and expansion of Intensive Care Units, emphasizing the role of intensivists in these cases.


title: Is radiofrequency ablation of varicose veins a valuable option? A systematic review of the literature with a cost analysis.

text: BACKGROUND Since the 1990s, new techniques for the treatment of varicose veins have emerged, including radiofrequency ablation (RFA) and laser treatment. We performed a study to compare the safety, efficacy and outcomes of RFA compared to those of open surgery and laser ablation for the treatment of varicose veins. We also carried out a cost analysis of RFA compared to open surgery to assess whether RFA could help free up operating room time by being performed in an outpatient context. METHODS We conducted a systematic literature review (publication date May 2010-September 2013 for articles in English, January 1991-September 2013 for those in French). We used several checklists to measure the quality of the studies. We also collected data on costing. RESULTS The literature search identified 924 publications, of which 38 were retained for analysis: 15 literature reviews, 1 good-practice guideline and 22 new primary studies. The overall level of evidence was low to moderate owing to the limited sample sizes, lack of information on patient characteristics and lack of standardization of the outcome measures. However, the results obtained are consistent from study to study. In the short and medium term, RFA is considered as effective as open surgery or laser treatment (moderate level of evidence) and presents fewer major and minor complications than open surgery (low level of evidence). Radiofrequency ablation can be performed on an outpatient basis. We calculated that RFA would be about $110-$220 more expensive per patient than open surgery. CONCLUSION Radiofrequency ablation is a valuable alternative to open surgery and would free up operating room time in a context of low accessibility.


title: Update: severe acute respiratory syndrome--Toronto, Canada, 2003.

text: Severe acute respiratory syndrome (SARS) was first recognized in Toronto in a woman who returned from Hong Kong on February 23, 2003 (1). Transmission to other persons resulted subsequently in an outbreak among 257 persons in several Greater Toronto Area (GTA) hospitals. After implementation of provincewide public health measures that included strict infection-control practices, the number of recognized cases of SARS declined substantially, and no cases were detected after April 20. On April 30, the World Health Organization (WHO) lifted a travel advisory issued on April 22 that had recommended limiting travel to Toronto. This report describes a second wave of SARS cases among patients, visitors, and health-care workers (HCWs) that occurred at a Toronto hospital approximately 4 weeks after SARS transmission was thought to have been interrupted. The findings indicate that exposure to hospitalized patients with unrecognized SARS after a provincewide relaxation of strict SARS control measures probably contributed to transmission among HCWs. The investigation underscores the need for monitoring fever and respiratory symptoms in hospitalized patients and visitors, particularly after a decline in the number of reported SARS cases.


title: [Reconstructive surgery protocol recommendations during COVID-19 pandemia.]

text: OBJECTIVES Offer some recommendations or guidelines during the evolution of the COVID-19 pandemic in terms of diagnosis, treatment and follow-upin the field of Reconstructive Urology. MATERIAL AND METHOD: The document is based on the evidence on SARS/Cov-2 and the authors' experience in managing COVID-19 in their institutions, including specialists from Andalusia, Madrid, Cantabria,the Valencian Community and Catalonia. A web and PubMed search was performed using "SARS-CoV-2", "COVID-19", "COVID-19 Urology", "COVID19 urology complications", "COVID-19 reconstructive surgery".A narrative review of the literature was carried out (5/17/2020) and after the nominal group technique modified due to the extraordinary restrictions, a first draft was made to unify criteria and reach a quick consensus. Finally, a definitive version was made, agreed by all the authors (5/22/2020). RESULTS: The authors defined the following surgical priorities for Urological Reconstructive Surgery: Emergency/Urgency (life-threatening or emergencies still in anormal situation), Elective Urgency/High priority (potentially dangerous pathology if postponed for more than 1month), Elective Surgery/Intermediate priority (pathology with little probability of being dangerous but it is recommended not to delay more than 6 months), Delayed surgery/Low priority (non-dangerous pathology if it is postponed for more than 6 months). According to this classification, the Working Group agreed on the distribution of the different surgical scenarios of Reconstructive Urology. In addition, consensus was reached on recommendations regarding the diagnosis and follow-up of pathology in the field of Reconstructive Urology. CONCLUSIONS: Tools should be implemented to facilitate the gathering of the medical visit and diagnostic tests. Redistribution of surgical procedures based on priority degrees is necessary during the pandemic and transition period. The use of telemedicine is essential forfollow-up, by computer, telephone or videoconference.


title: Involvement of aminopeptidase N (CD13) in infection of human neural cells by human coronavirus 229E.

text: Attachment to a cell surface receptor can be a major determinant of virus tropism. Previous studies have shown that human respiratory coronavirus HCV-229E uses human aminopeptidase N (hAPN [CD13]) as its cellular receptor for infection of lung fibroblasts. Although human coronaviruses are recognized respiratory pathogens, occasional reports have suggested their possible neurotropism. We have previously shown that human neural cells, including glial cells in primary cultures, are susceptible to human coronavirus infection in vitro (A. Bonavia, N. Arbour, V. W. Yong, and P. J. Talbot, J. Virol. 71:800-806, 1997). However, the only reported expression of hAPN in the nervous system is at the level of nerve synapses. Therefore, we asked whether hAPN is utilized as a cellular receptor for infection of these human neural cell lines. Using flow cytometry, we were able to show the expression of hAPN on the surfaces of various human neuronal and glial cell lines that are susceptible to HCV-229E infection. An hAPN-specific monoclonal antibody (WM15), but not control antibody, inhibited the attachment of radiolabeled HCV-229E to astrocytic, neuronal, and oligodendrocytic cell lines. A correlation between the apparent amount of cell surface hAPN and the level of virus attachment was observed. Furthermore, the presence of WM15 inhibited virus infection of these cell lines, as detected by indirect immunofluorescence. These results indicate that hAPN (CD13) is expressed on neuronal and glial cell lines in vitro and serves as the receptor for infection by HCV-229E. This further strengthens the neurotropic potential of this human respiratory virus.


title: Manipulation of immunity to and pathology of respiratory infections.

text: Respiratory infections are the third leading cause of death worldwide and are a priority for vaccine development. Immune defence mechanisms are critical in recovery from most respiratory infections but the role of the immune system in causing bystander lung injury is not as well understood, and will be the focus of this review. Immune-mediated injury results from physical occlusion of the airways or the ensuing 'cytokine storm', which may spill over into the systemic circulation and cause devastating consequences. Respiratory pathogens employ numerous strategies to avoid detection by the immune system. One of these, the alteration of key surface determinants, makes the design of rational vaccines problematic. In the following review the immune compartments responsible for clinical lung disease are discussed, and current and novel strategies to reduce their potency are overviewed.


The Promise and Challenge of Home Health Services During the COVID-19 Pandemic.


title: Mapping sequences active in homologous RNA recombination in brome mosaic virus: prediction of recombination hot spots.

text: The mechanism of homologous recombination has been studied previously in brome mosaic virus (BMV), a tricomponent, positive-stranded RNA virus of plants, by using artificial sequences (reviewed by J. J. BujarskiP. D. Nagy (1996). Semin. Virol. 7, 363-372). Here we extend these studies over BMV-derived sequences to obtain clues on prediction of homologous recombination hot spots. First, mismatch mutations, which reduced the AU content, were introduced into the common 60-nt recombination hot-spot sequence, either in the RNA2 or in both RNA2RNA3 components. This decreased the frequency of targeted homologous RNA2/RNA3 recombinationchanged the distribution of junction sites. Second, several short BMV RNA1- or RNA2-derived sequences were introduced into the RNA3 component, homologous recombination activity of these sequences was compared with that observed for previously characterized artificial sequences. Third, sequences at homologous recombinant junctions were compared among a large number of targetednontargeted recombinants. All these studies revealed several factors important for homologous recombination including the length of sequence identity, the extent of sequence identity, the AU content of the common sequences, the relative position of the AU-rich segment vs a GC-rich segment,the presence of GC-rich sequences. Based on this novel model, we suggest that recombination hot spots can be predicted by means of RNA sequence analysis. In addition, we show that recombination can occur between positivenegative strands of BMV RNAs. This provides further clues toward the mechanism of recombination processes in BMV.


title: Prevalence of regular physical activity among adults--United States, 2001 and 2005.

text: Regular physical activity is associated with decreased risk for obesity, heart disease, hypertension, diabetes, certain cancers, and premature mortality. CDC and the American College of Sports Medicine recommend that adults engage in at least 30 minutes of moderate physical activity on most days and preferably on all days. Healthy People 2010 objectives include increasing the proportion of adults who engage regularly in moderate or vigorous activity to at least 50% (objective 22-2). In addition, reducing racial and ethnic health disparities, including disparities in physical activity, is an overarching national goal. To examine changes in the prevalence of regular, leisure-time, physical activity from 2001 to 2005, CDC analyzed data from the Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the results of that analysis, which indicated that, from 2001 to 2005, the prevalence of regular physical activity increased 8.6% among women overall (from 43.0% to 46.7%) and 3.5% among men (from 48.0% to 49.7%). In addition, the prevalence of regular physical activity increased 15.0% (from 31.4% to 36.1%) among non-Hispanic black women and 12.4% (from 40.3% to 45.3%) among non-Hispanic black men, slightly narrowing previous racial disparities when compared with increases of 7.8% (from 46.0% to 49.6%) for white women and 3.4% (from 50.6% to 52.3%) for white men, respectively. CDC, state and local public health agencies, and other public health partners should continue to implement evidence-based, culturally appropriate initiatives to further increase physical-activity levels among all adults, with special focus on eliminating racial/ethnic disparities.


Amplified fragment length polymorphism differentiation between the vaccine strain Chlamydia psittaci 1B and wild field strains.


title: Serologic survey of selected viral agents in recently captured wild North American river otters (Lontra canadensis).

text: Blood samples were collected from 64 wild North American river otters (Lontra [Lutra] canadensis) from northern and eastern New York State and analyzed for serologic evidence of exposure to selected viral agents during a 1995 1996 translocation program. No clinical signs of disease nor lesions suggestive of prior viral exposure were seen. Titers were detected for antibodies against canine distemper virus, canine herpesvirus-1, and canine parvovirus-2 but not for antibodies against canine adenovirus-1, canine coronavirus, canine parainfluenza virus, rabies virus, feline herpesvirus-1, feline calicivirus, or feline coronavirus. This is the first report of titers for antibodies against canine herpesvirus-1 in North American river otters, and it suggests a low prevalence of antibody titers against most canine viruses in otter populations in northern and eastern New York. Confounding variables in this study could include exposure to domestic dogs associated with the project, prolonged time spent in captivity, and concurrent bacterial or parasitic infection. Stress-associated humoral immune suppression could have altered serologic profiles, especially in otters exposed to dogs after trapping but before venipuncture.


title: [Validation of surgical masks during COVID19 emergency: activities at the University of Napoli Federico II].

text: SUMMARY During COVID-19 pandemic crisis, Italian Government has approved Law Decree no. 18 of 17 march 2020, in which art. 15 allows enterprises to produce, import and commercialize surgical masks notwithstanding the current rules of product certification. It is just required that the interested enterprises send to the Italian National Institute of Health a selfcertification in which they declare the technical characteristics of the masks and that masks are produced according to the safety requirements. In this context, a technical-scientific unit was established at the University of Napoli Federico II to provide interested enterprises with state-of-the-art consultancy, testing and measurement services, adhering to rigorous scientific protocols. Characterization tests were carried out on 163 surgical masks and/or materials for their construction and they have enabled the identification of pre-screening criteria to simplify the procedure for evaluating surgical masks using methods for assessing the filtration efficiency of particles and aerosols. Based on experimental results, it has been observed that a filtration efficiency for particles with sizes larger that 650 nm (PFE>650) exceeding 35% might guarantees a bacterial filtration efficiency (BFE) higher than 95% while BFE values higher than 98% are obtained when the PFE>650 is larger than 40%. PFE measurement is extremely simpler with respect to BFE, the latter being time-consuming and requiring specific equipment and methods for its realization. Many tested materials have shown the capability to assure high filtration efficiencies but Spundonded-Meltblown-Spunbonded (SMS), that are layers of non-woven fabric with different weights of Meltblown, can simultaneously guarantee high particle filtration efficiencies with pressure drop values (breathability) in the limits to classify the surgical masks as Type II/IIR. In fact, the fabric products analyzed so far have not been able to simultaneously guarantee adequate BFE and breathability values. On the contrary, Spunbonds of adequate weights can virtually verify both requirements and accredit themselves as possible materials for the production of surgical masks, at least of Type I. Further studies are needed to verify the possibility of producing low-cost, reusable surgical masks that could meet the criteria of circular economy.


Public health education: The use of unique strategies to educate the public in the principles of the Hartford Consensus.


rTMS as an add-on treatment for resistant obsessive-compulsive symptoms in patients with schizophrenia: report of three cases.


Could Demodex Be Used to Augment a Topical Vaccine Against Coronavirus (and Similar Viruses)?


Sleeping sickness--a growing problem?


Boosting the sensitivity of real-time polymerase-chain-reaction testing for SARS.


Ribavirin in the treatment of SARS: A new trick for an old drug?


Revision of the International Health Regulations.


title: Recombination between nonsegmented RNA genomes of murine coronaviruses.

text: We have isolated a recombinant virus between the A59 and JHM strains of mouse hepatitis virus, which contain a single species of nonsegmented RNA genome. This recombinant was derived by mixed infection of DBT cells with temperature-sensitive mutants of A59 and JHM at nonpermissive temperature. Viruses recovered at this temperature were screened by oligonucleotide fingerprinting of their genomic RNAs. One recombinant virus, B1, was found to contain mostly A59-derived sequences, but the 3 kilobases at the 5' end of the genomic RNA was derived from JHM. Thus, the crossover point in the B1 genome is located within gene A, which codes for the viral RNA polymerases. The study of the intracellular RNA species of B1 virus revealed that probably all of the virus-specific subgenomic mRNA species contained the body sequences of strain A59 but the leader sequences of JHM. This result indicates that the JHM leader RNA, which differs from the A59 leader RNA, could be fused to the mRNAs of a different virus strain during RNA transcription. Furthermore, B1 virus-infected cells contain an additional subgenomic mRNA species which is transcribed from a new initiation site within gene C, suggesting that the leader RNA could determine the site of initiation for coronavirus mRNAs. These data represent a first report of RNA recombination between viruses, other than picornaviruses, which contain nonsegmented RNA genomes.


title: Severe acute respiratory syndrome: temporal lung changes at thin-section CT in 30 patients.

text: PURPOSE To evaluate lung abnormalities on serial thin-section computed tomographic (CT) scans in patients with severe acute respiratory syndrome (SARS) during acute and convalescent periods. MATERIALS AND METHODS Serial thin-section CT scans in 30 patients (17 men, aged 42.5 years +/- 12.2 [SD]) with SARS were reviewed by two radiologists together for predominant patterns of lung abnormalities: ground-glass opacities, ground-glass opacities with superimposed linear opacities, consolidation, reticular pattern, and mixed pattern (consolidation, ground-glass opacities, and reticular pattern). Scans were classified according to duration in weeks after symptom onset. Longitudinal changes of specific abnormalities were documented in 17 patients with serial scans obtained during 3 weeks. Each lung was divided into three zones; each zone was evaluated for percentage of lung involvement. Summation of scores from all six lung zones provided overall CT score (maximal CT score, 24). RESULTS Median CT scores increased from 1 in the 1st week to 12.5 in the 2nd week. Ground-glass opacities with or without smooth interlobular septal thickening and consolidation were predominant patterns found during the 1st week. Ground-glass opacities with superimposed irregular reticular opacities, mixed pattern, and reticular opacities were noted from the 2nd week and peaked at or after the 4th week. After the 4th week, 12 (55%) of 22 patients had irregular linear opacities with or without associated ground-glass opacities and CT scores greater than 5; five of these patients had bronchial dilatation. When specific opacities were analyzed in 17 patients, consolidation generally resolved completely (n = 4) or to minimal residual opacities; six (55%) of 11 patients with ground-glass opacities had substantial residual disease (CT scores > 5) on final scans. CONCLUSION There is a temporal pattern of lung abnormalities at thin-section CT in SARS. Predominant findings at presentation are ground-glass opacities and consolidation. Reticulation is evident after the 2nd week and persists in half of all patients evaluated after 4 weeks. Long-term follow-up is required to determine whether the reticulation represents irreversible fibrosis.


title: Influenza infection screening tools fail to accurately predict influenza status for patients during pandemic H1N1 influenza season.

text: BACKGROUND Following the severe acute respiratory syndrome outbreak in 2003, hospitals have been mandated to use infection screening questionnaires to determine which patients have infectious respiratory illness and, therefore, require isolation precautions. Despite widespread use of symptom-based screening tools in Ontario, there are no data supporting the accuracy of these screening tools in hospitalized patients. OBJECTIVE To measure the performance characteristics of infection screening tools used during the H1N1 influenza season. METHODS The present retrospective cohort study was conducted at The Ottawa Hospital (Ottawa, Ontario) between October and December, 2009. Consecutive inpatients admitted from the emergency department were included if they were ≥18 years of age, underwent a screening tool assessment at presentation and had a most responsible diagnosis that was cardiac, respiratory or infectious. The gold-standard outcome was laboratory diagnosis of influenza. RESULTS The prevalence of laboratory-confirmed influenza was 23.5%. The sensitivity and specificity of the febrile respiratory illness screening tool were 74.5% (95% CI 60.5% to 84.8%) and 32.7% (95% CI 25.8% to 40.5%), respectively. The sensitivity and specificity of the influenza-like illness screening tool were 75.6% (95% CI 61.3% to 85.8%) and 46.3% (95% CI 38.2% to 54.7%), respectively. CONCLUSIONS The febrile respiratory illness screening tool missed 26% of active influenza cases, while 67% of noninfluenza patients were unnecessarily placed under respiratory isolation. Results of the present study suggest that infection-control practitioners should re-evaluate their strategy of screening patients at admission for contagious respiratory illness using symptom- and sign-based tests. Future efforts should focus on the derivation and validation of clinical decision rules that combine clinical features with laboratory tests.


title: SARS Surveillance Project--Internet-enabled multiregion surveillance for rapidly emerging disease.

text: INTRODUCTION On March 15, 2003, CDC requested health-care and public health agencies to conduct surveillance for severe acute respiratory syndrome (SARS). The SARS Surveillance Project (SARS-SP) was established to rapidly implement multiregional SARS surveillance in emergency departments (EDs) by using existing Internet-based tools. OBJECTIVES The objectives of SARS-SP were to 1) disseminate and update SARS screening forms for ED triage, 2) establish surveillance for SARS syndrome elements by using Regional Emergency Medicine Internet (REMI), 3) expand surveillance to multiple regions, and 4) evaluate the usefulness of Internet tools for agile surveillance during a rapidly emerging global epidemic. METHODS SARS-SP developed, distributed, and updated an Internet-based triage form to identify patients for infection control and public health reporting. EDs then were invited to report visit frequencies with various SARS syndrome elements to local public health authorities by using the REMI Internet application (first in one metropolitan area, and later in four). After pilot-testing in one metropolitan area, the surveillance system was implemented in three others. RESULTS Active syndromic surveillance was established by health departments in Milwaukee, Wisconsin; Denver, Colorado; Akron, Ohio; and Fort Worth, Texas. A total of 27 EDs reported syndrome frequencies from >146,000 patient encounters. CONCLUSIONS ED and public health partners reported being satisfied with the system, confirming the usefulness of Internet tools in the rapid establishment of multiregion syndromic surveillance during an emerging global epidemic.


title: Exposure to infectious agents in dogs in remote coastal British Columbia: Possible sentinels of diseases in wildlife and humans.

text: Ranked among the top threats to conservation worldwide, infectious disease is of particular concern for wild canids because domestic dogs (Canis familiaris) may serve as sources and reservoirs of infection. On British Columbia's largely undeveloped but rapidly changing central and north coasts, little is known about diseases in wolves (Canis lupus) or other wildlife. However, several threats exist for transfer of diseases among unvaccinated dogs and wolves. To gain baseline data on infectious agents in this area, including those with zoonotic potential, we collected blood and stool samples from 107 dogs in 5 remote communities in May and September 2007. Serology revealed that the dogs had been exposed to canine parvovirus, canine distemper virus, Bordetella bronchiseptica, canine respiratory coronavirus, and Leptospira interrogans. No dogs showed evidence of exposure to Ehrlichia canis, Anaplasma phagocytophilum, Borrelia burgdorferi, Dirofilaria immitis, or Cryptococcus gattii. Of 75 stool samples, 31 contained at least 1 parasitic infection, including Taeniid tapeworms, the nematodes Toxocara canis and Toxascaris leonina, and the protozoans Isospora sp., Giardia sp., Cryptosporidium sp., and Sarcocystis sp. This work provides a sound baseline for future monitoring of infectious agents that could affect dogs, sympatric wild canids, other wildlife, and humans.


title: The impact of SARS on a tertiary care pediatric emergency department.

text: BACKGROUND The Greater Toronto Area (GTA) was considered a "hot zone" for severe acute respiratory syndrome (SARS) in 2003. In accordance with mandated city-wide infection control measures, the Hospital for Sick Children (HSC) drastically reduced all services while maintaining a fully operational emergency department. Because of the GTA health service suspensions and the overlap of SARS-like symptoms with many common childhood illnesses, this introduced the potential for a change in the volumes of patients visiting the emergency department of the only regional tertiary care children's hospital. METHODS We compared HSC emergency department patient volumes, admission rates and length of stay in the emergency department in the baseline years of 2000-2002 (non-SARS years) with those in 2003 (SARS year). The data from the prior years were modeled as a time series. Using an interrupted time series analysis, we compared the 2003 data for the periods before, during and after the SARS periods with the modeled data for significant differences in the 3 aforementioned outcomes of interest. RESULTS Compared with the 2000-2002 data, we found no differences in visits, admission rates or length of stay in the pre-SARS period in 2003. There were significant decreases in visits and length of stay (p < 0.001) and increases in admission rates (p < 0.001) during the periods in 2003 when there were new and active cases of SARS in the GTA. All 3 outcomes returned to expected estimates coincident with the absence of SARS cases from September to December 2003. INTERPRETATION During the SARS outbreak in the GTA, the HSC emergency department experienced significantly reduced volumes of patients with low-acuity complaints. This gives insight into utilization rates of a pediatric emergency department during a time when there was additional perceived risk in using emergency department services and provides a foundation for emergency department preparedness policies for SARS-like public health emergencies.


title: Vaccines based on novel adeno-associated virus vectors elicit aberrant CD8+ T-cell responses in mice.

text: We recently discovered an expanded family of adeno-associated viruses (AAVs) that show promise as improved gene therapy vectors. In this study we evaluated the potential of vectors based on several of these novel AAVs as vaccine carriers for human immunodeficiency virus type 1 Gag. Studies with mice indicated that vectors based on AAV type 7 (AAV7), AAV8, and AAV9 demonstrate improved immunogenicity in terms of Gag CD8(+) T-cell and Gag antibody responses. The quality of these antigen-specific responses was evaluated in detail for AAV2/8 vectors and compared to results with an adenovirus vector expressing Gag (AdC7). AAV2/8 produced a vibrant CD8(+) T-cell effector response characterized by coexpression of gamma interferon and tumor necrosis factor alpha as well as in vivo cytolytic activity. No CD8(+) T-cell response generated by any of the AAVs was effectively boosted with AdC7, a result consistent with the finding of a relative lack of cells expressing interleukin-2 (IL-2) or a central memory phenotype at 3 months after the prime. The primary response to an AdC7 vaccine differed from that generated by AAVs in that the peak effector response evolved into populations of Gag-specific T cells expressing high levels of cytokines, including IL-2, and with effector memory and central memory phenotypes. A number of mechanisms could be considered to explain the aberrant activation of CD8(+) T cells by AAV, including insufficient inflammatory responses, CD4 help, and/or chronic antigen expression and T-cell exhaustion. Interestingly, the B-cell response to AAV-encoded Gag was quite vibrant and easily boosted with AdC7.


title: [Recommendations of the Society of Gastroenterology of Peru to avoid the spread of SARS-CoV-2 through digestive endoscopy procedures].

text: The SARS-CoV-2 coronavirus produces the disease called COVID-19, currently spreading in a rapidly evolving pandemic. It can be transmitted by contact, drops and aerosols, and has been isolated from gastrointestinal secretions and faeces. During digestive endoscopy, transmission by any of these mechanisms could occur. It is recommended to limit digestive endoscopy to cases of digestive bleeding, severe dysphagia, foreign body in the digestive tract, biliary obstruction with intractable pain or cholangitis, pseudocyst or complicated encapsulated pancreatic necrosis, gastrointestinal obstruction, and cases at risk of deterioration over time. It is recommended to screen patients based on temperature, symptoms, and epidemiological factors to classify them according to their risk of infection. For procedures in low risk patients, personnel must wear a disposable gown, gloves, eye or face shield, standard surgical mask, disposable hat, disposable shoe covers. In cases of intermediate or high risk, or confirmed COVID-19, protection should be increased using disposable waterproof gown, N95 respirator or similar, and double glove. In case of shortage it may be necessary to reuse N95 respirators for up to 5 uses, following CDC recommendations for donning, removing and storing to prevent secondary contact contamination. Likewise, all protective equipment should be put on and removed according to CDC recommendations. The presence of personnel in endoscopy should be limited to the bare minimum. Said personnel must have daily temperature control and if it is above 37.3ºC, the corresponding evaluation must be carried out. After each procedure, the stretcher and room surfaces should be properly disinfected. High-level disinfection of endoscopes eliminates SARS-CoV-2.


title: An Improved Surgical Technique for Pure Laparoscopic Left Hemihepatectomy: Ten Years Experience in a Tertiary Center.

text: BACKGROUND This study details our experience with an improved surgical technique involving the hepatic pedicle during laparoscopic left hemihepatectomy (LLH). METHODS We describe an improved laparoscopic technique to extraparenchymally divide the left hepatic pedicle. A retrospective analysis of all of the patients who underwent laparoscopic liver procedures between 2002 and 2012 was conducted. The patients were divided into two groups, an early LLH group (ELLH group) and a recent LLH group (RLLH group), based on the surgical approach used for the left hepatic pedicle. RESULTS A total of 72 cases of LLH (26 ELLH and 46 RLLH) were identified. The RLLH group exhibited a shorter median operative time, median length of hospital stay, and lower median blood loss compared to the ELLH group (182, 162.5-223.7 versus 232.5, 200-357.5 minutes, P < .01; 5, 4.2-7 versus 7, 6-8.7 days, P < .05; 150, 100-257.5 versus 300, 200-337.5 mL, P < .05, respectively). No perioperative mortality was observed. CONCLUSIONS This study confirms that our improved surgical technique for LLH is practical, safe, and effective. The main advantage of this method compared to other techniques is the possibility of attaining rapid and precise control of vascular inflow, thus facilitating LLH.


title: Comparison of three ventilatory modes during one-lung ventilation in elderly patients.

text: OBJECTIVE The purpose of this study was to explore the effects of three different ventilatory modes: volume controlled ventilation (VCV), pressure controlled ventilation (PCV) and pressure controlled ventilation-volume guaranteed (PCV-VG) on arterial oxygenation and airway pressure during one-lung ventilation (OLV) in elderly patients. METHODS We enrolled 66 patients who underwent thoracic surgery requiring at least 1 hour of OLV and aged above 65 years into the study. Patients were classified into VCV, PCV and PCV-VG groups according to a controlled, randomized design. Patients were ventilated to obtain a tidal volume (TV) of 8 mL/kg with three different ventilatory modes during OLV. The Hemodynamic and respiratory data had been recorded during intraoperation and arterial blood gases were obtained at baseline, 20, 40, 60 minutes after OLV, end of surgery. RESULTS Compared with VCV group, Ppeak was significantly lower in PCV and PCV-VG group (P<0.05), and the difference was not found between the PCV and PCV-VG group. PaO2 in PCV and PCV-VG group were higher than VCV group after the point of OLV+40 (P<0.05). Comparison of PCV group, PaO2 in PCV-VG group was higher, but did not show a significantly improved during OLV (P>0.05). CONCLUSIONS Compared with VCV, the use of PCV and PCV-VG have a significant advantage in intraoperative oxygenation and airway pressure for eldly patients undergoing OLV.


title: False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid enzyme-linked immunosorbent assay due to HCoV-OC43 and HCoV-229E rectified by Western blotting with recombinant SARS-CoV spike polypeptide.

text: Using paired serum samples obtained from patients with illness associated with increases in anti-human coronavirus OC43 (HCoV-OC43) or anti-HCoV-229E antibodies, we examined the possibility of false-positive results detected in a recombinant severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) nucleocapsid protein immunoglobulin G enzyme-linked immunosorbent assay (ELISA). Three of the 21 and 1 of the 7 convalescent-phase serum samples from persons with increases in antibodies against HCoV-OC43 and HCoV-229E, respectively, tested positive by the recombinant SARS-CoV nucleocapsid protein-based ELISA. None of these samples were found to contain a specific antibody in the recombinant SARS-CoV spike polypeptide-based Western blot assay.


title: Cognitive rehabilitation in the elderly: a randomized trial to evaluate a new protocol.

text: This study provides an introduction to, and overview of, several papers that resulted from a randomized control trial that evaluated a new cognitive rehabilitation protocol. The program was designed to improve general strategic abilities in ways that would be expressed in a broad range of functional domains. The trial, which was conducted on a sample of older adults who had experienced normal age-related cognitive decline, assessed performance in the following domains: memory, goal management, and psychosocial status. The general rationale for the trial, the overall experimental design, and the approach to statistical analyses that are relevant to each paper are described here. The results for each functional domain are reported in separate papers in this series.


title: The cytoplasmic tail of infectious bronchitis virus E protein directs Golgi targeting.

text: We have previously shown that the E protein of the coronavirus infectious bronchitis virus (IBV) is localized to the Golgi complex when expressed exogenously from cDNA. Here, we report that neither the transmembrane domain nor the short lumenal domain of IBV E is required for Golgi targeting. However, an N-terminal truncation containing only the cytoplasmic domain (CTE) was efficiently localized to the Golgi complex, and this domain could retain a reporter protein in the Golgi. Thus, the cytoplasmic tail of the E protein is necessary and sufficient for Golgi targeting. The IBV E protein is palmitoylated on one or two cysteine residues adjacent to its transmembrane domain, but palmitoylation was not required for proper Golgi targeting. Using C-terminal truncations, we determined that the IBV E Golgi targeting information is present between tail amino acids 13 and 63. Upon treatment with brefeldin A, both the E and CTE proteins redistributed to punctate structures that colocalized with the Golgi matrix proteins GM130 and p115 instead of being localized to the endoplasmic reticulum like Golgi glycosylation enzymes. This suggests that IBV E is associated with the Golgi matrix through interactions of its cytoplasmic tail and may have interesting implications for coronavirus assembly in early Golgi compartments.


title: Glycyrrhizin inhibits R5 HIV replication in peripheral blood monocytes treated with 1-methyladenosine.

text: R5 HIV replicated in freshly isolated human peripheral blood monocytes after treatment with 1-methyladenosine (fresh PBM/MA), an immunosuppressive compound isolated from tumorous ascites fluids, while viral replication was not demonstrated in untreated peripheral blood monocytes (fresh PBM). The R5 HIV replication in fresh PBM/MA was inhibited by glycyrrhizin (GL). Without any other stimulation, fresh PBM/MA produced CCL2 and IL-10, while these soluble factors were not released from fresh PBM. GL greatly inhibited the production of CCL2 and IL-10 in fresh PBM/MA. After treatment with CCL2 and/or IL-10, CCR5 mRNA expression in fresh PBM was markedly enhanced, while only a trace level of the mRNA expression was detected in these cells in the absence of CCL2 or IL-10. CCR5 mRNA expression in fresh PBM treated with CCL2 or IL-10 was clearly inhibited by GL. These results indicate that GL inhibits R5 HIV replication in fresh PBM/MA through the inhibiting CCR5 expression mediated by CCL2 or IL-10.


Literature.


title: Microbiology of Community-Acquired Pneumonia in the Gulf Corporation Council States.

text: In spite of advances in microbiological and serological investigations over the last two decades, etiological attribution remains difficult in community-acquired pneumonia (CAP). Even after exhaustive investigation, the etiology of CAP remains unknown in up to 50% of patients. Common pathogens include Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis. In addition, several investigators document the importance of atypical pathogens including Mycoplasma pneumoniae, Chlamydophila pneumoniae and Legionella pneumophila in the etiology of CAP in the GCC region. Increasingly, other etiologies, particularly influenza viruses, varicella zoster virus and Mycobacterium tuberculosis, have been recognized as causative pathogens of CAP within the region. Rates of antimicrobial resistance of S. pneumoniae and other pathogens are rising in the Gulf Corporation Council (GCC) region and susceptibility profiles of antibiotics against intracellular pathogens such as Chlamydophila pneumoniae and Mycoplasma pneumoniae are not routinely performed. Injudicious prescribing and over-use of antibiotics drive much resistance. The GCC CAPWG calls for urgent governmental regulations to limit and monitor antibiotic prescription in the GCC region.


title: Two New Alkaloids from the Roots of Baphicacanthus cusia.

text: Phytochemical investigation of the root of Baphicacanthus cusia (NEES) BREMEK afforded two new alkaloids, baphicacanthin A (1) and baphicacanthin B (2), along with 28 known compounds. The chemical structures of these compounds were elucidated on the basis of one and two dimensional (1D/2D)-NMR and high resolution (HR)-MS spectral evidence.


title: [The specific features of the reproduction of the bovine coronavirus strain ARRIAH in the continuous RBT cell culture].

text: Cultivation of the bovine coronavirus strain ARRHIAH in the continuous RBT versus MDBK and Taurus cell cultures made it possible to reduce the time taken to reproduce the virus and to increase its hemagglutination titer. The RBT cell line is suitable for determining the infectious activity of the coronavirus reproduced in the other cell cultures. Optimization of a manufacturing process for diagnostic agents and vaccines has been achieved while applying a roll tube method for both RBT cell cultivation and coronavirus reproduction.


title: Single-incision videolaparoscopic appendectomy with conventional videolaparoscopy equipment.

text: BACKGROUND Acute appendicitis is the most common surgical emergency in daily practice, and is approached laparoscopically in many centers. Efforts have been undertaken for the development of minimally invasive techniques that reduce tissue trauma and offer improved cosmetic results, one of such being the single-incision laparoscopic surgery (SILS). AIM To present a minimally invasive technique for appendectomy (SILS) undertaken with conventional instruments. METHOD Eleven patients were treated in the emergency care center presenting abdominal pain in the right iliac fossa that was suggestive of appendicitis. Diagnostic investigation was subsequently conducted, including physical examination, laboratory and imaging exams (CT scan with intravenous contrast or total abdominal ultrasound), and the results were consistent with acute appendicitis. Thus, after consent, these patients underwent SILS appendectomy under general anesthesia with three trocars (two 10 mm and one 5 mm), using conventional and optical laparoscopic tweezers (10 mm, 30º). The base and pedicle of the appendix were ligated with titanium LT 400 clips. The procedure occurred uneventfully. Inclusion criteria were absence of diffuse peritonitis, BMI (body mass index) less than 35 and absence of serious comorbidities or sepsis. RESULTS Seven men and four women were operated with average age of 25.7 years and underwent appendectomy through this technique. Mean procedure duration was of 37.2 min. Regarding surgical findings, three had appendicitis in stage 1, four in stage 2 and four in stage 3. All patients improved well, without surgical complications, and did not require conversion to open surgery or conventional laparoscopy technique. CONCLUSION Appendectomy conducted through Single Incision Laparoscopic Surgery is a feasible and promising technique that can be performed with conventional laparoscopic instruments.


MSMA Urges Congress Independent Physician Financial Stability in Economic COVID-19 Relief Legislation.


title: Severe acute respiratory syndrome coronavirus pathogenesis, disease and vaccines: an update.

text: BACKGROUND A novel coronavirus has recently been identified as the cause of severe acute respiratory syndrome (SARS-CoV). The ability of this family of positive strand RNA viruses to move between species and cause severe disease in humans, with the potential for pandemic spread, has been confirmed. METHODS An understanding of the disease and its pathogenesis and the genetics of coronavirus infections, as well as strategies to treat or prevent coronavirus infections, are essential. The history of coronavirus vaccines and the occurrence of laboratory-associated SARS-CoV infections underscore the need for stably attenuated strains of SARS-CoV and other coronaviruses. RESULTS Rapid progress has been made in understanding the clinical disease of SARS in adults and children. In adults, systemic infection with clinical and biochemical abnormalities, as well as respiratory infection, may be the rule. SARS is much milder in children younger than 12 years old than it is in adolescents and adults. In children age 12 years and younger, symptoms are generally nonspecific and cold-like. Numerous approaches to the development of SARS-CoV vaccines have been undertaken, and there is evidence that antibodies to the spike protein may be protective from replication and pathology in animal models. CONCLUSIONS The availability of reverse genetic systems has made it possible to engineer and recover coronavirus variants that contain multiple genetically stable mutations that grow well in culture but are attenuated for replication, virulence or both. Such variants will be platforms for the safe growth of SARS-CoV and candidates for live attenuated vaccines.


Hemadsorption by coronavirus strain OC43.


title: Acute respiratory distress syndrome in critically ill patients with severe acute respiratory syndrome.

text: CONTEXT Severe acute respiratory syndrome (SARS) is an emerging infectious disease with a 25% incidence of progression to acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) and mortality exceeding 10%. OBJECTIVE To describe the clinical spectrum and outcomes of ALI/ARDS in patients with SARS-related critical illness. DESIGN, SETTING, AND PATIENTS Retrospective case series of adult patients with probable SARS admitted to the intensive care unit (ICU) of a hospital in Singapore between March 6 and June 6, 2003. MAIN OUTCOME MEASURES The primary outcome measure was 28-day mortality after symptom onset. RESULTS Of 199 patients hospitalized with SARS, 46 (23%) were admitted to the ICU, including 45 who fulfilled criteria for ALI/ARDS. Mortality at 28 days for the entire cohort was 20 (10.1%) of 199 and for ICU patients was 17 (37%) of 46. Intensive care unit mortality at 13 weeks was 24 (52.2%) of 46. Nineteen of 24 ICU deaths occurred late (> or =7 days after ICU admission) and were attributed to complications related to severe ARDS, multiorgan failure, thromboembolic complications, or septicemic shock. ARDS was characterized by ease of derecruitment of alveoli and paucity of airway secretion, bronchospasm, or dynamic hyperinflation. Lower Acute Physiology and Chronic Health Evaluation II scores and higher baseline ratios of PaO2 to fraction of inspired oxygen were associated with earlier recovery. CONCLUSIONS Critically ill patients with SARS and ALI/ARDS had characteristic clinical findings, high rates of complications; and high mortality. These findings may provide useful information for optimizing supportive care for SARS-related critical illness.


title: Genetic stability of equine arteritis virus during horizontal and vertical transmission in an outbreak of equine viral arteritis.

text: An imported carrier stallion (A) from Europe was implicated in causing an extensive outbreak of equine viral arteritis (EVA) on a Warmblood breeding farm in Pennsylvania, USA. Strains of equine arteritis virus (EAV) present in the semen of two carrier stallions (A and G) on the farm were compared to those in tissues of foals born during the outbreak, as well as viruses present in the semen of two other stallions that became persistently infected carriers of EAV following infection during the outbreak. The 2822 bp segment encompassing ORFs 2-7 (nt 9807-12628; which encode the G(S), GP3, GP4, G(L), M and N proteins, respectively) was directly amplified by RT-PCR from semen samples and foal tissues. Nucleotide and phylogenetic analyses confirmed that virus present in the semen of stallion A initiated the outbreak. The genomes of viruses present in most foal tissues (10/11) and serum from an acutely infected mare collected during the outbreak were identical to that of virus present in the lung of the first foal that died of EVA. Virus in the placenta of one foal differed by one nucleotide (99.9% identity) from the predominant outbreak virus. The relative genetic stability of viruses that circulated during the outbreak contrasts markedly with the heterogeneous virus populations variously present in the semen of persistently infected stallions on the farm. These findings are consistent with the hypothesis that the carrier stallion can be a source of genetic diversity of EAV, and that outbreaks of EVA can be initiated by the horizontal aerosol transmission of specific viral variants that occur in the semen of particular carrier stallions.


title: Reproduction of proventriculitis in commercial and specific-pathogen-free broiler chickens.

text: Proventriculitis was studied by experimentally reproducing the disease in broiler chickens. One-day-old infectious bursal disease virus (IBDV) and infectious bronchitis virus (IBV) antibody positive commercial broilers and 1-day-old antibody negative specific-pathogen-free (SPF) broilers were orally gavaged with proventricular homogenates produced from the proventriculi of broilers with proventriculitis. At 7 and 14 days, both commercial and SPF broilers had enlargement of the proventriculus with necrosis of the glandular epithelium and lymphocytic infiltrates in the proventricular glands. SPF broilers exposed to the proventricular homogenates developed infectious bursal disease, and IBDV was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). They also were positive by RT-PCR to IBV and developed nephritis. Commercial broilers developed mild nephritis but not bursal disease and were negative for IBDV and positive for IBV by RT-PCR. Both homogenate-inoculated commercial and SPF chickens were negative for reovirus and Newcastle disease virus by RT-PCR and variably positive for adenovirus by PCR. Bacteria were not identified in histologic sections, nor were they isolated from affected proventriculi. Indirect fluorescent antibody assay using convalescent sera detected intracytoplasmic staining in the proventricular glandular epithelial cells. Examination of thin sections of proventriculi using electron microscopy identified virus-like particles approximately 120 nm in diameter within the cytoplasm of these cells at 7 days after inoculation. Passage of proventricular homogenate filtrates in chicken embryos for virus isolation caused stunting, and allantoic fluid from these eggs was positive for IBV by RT-PCR.


title: Animal coronaviruses: what can they teach us about the severe acute respiratory syndrome?

text: In 2002, a new coronavirus (CoV) emerged in the People's Republic of China, associated with a severe acute respiratory syndrome (SARS) and mortality in humans. The epidemic rapidly spread throughout the world before being contained in 2003, although sporadic cases occurred thereafter in Asia. The virus is thought to be of zoonotic origin from a wild animal reservoir (Himalayan palm civets [Paguma larvata] are suspected), but the definitive host is unknown. There is concern about possible transmission of SARS CoV to rodents or domestic cats (as proven experimentally) with perpetuation of the disease in these species. In livestock and poultry, CoVs are recognised causes of enteric and respiratory infections that are often fatal in young animals. Although the emergence of SARS surprised the medical community, veterinary coronavirologists had previously isolated CoVs from wildlife and documented their interspecies transmission to livestock. Furthermore, scientists were aware of compelling evidence pointing to the emergence of new CoV strains and the mutation of existing strains resulting in new disease syndromes in animals, but the evolution and disease impact of CoVs was not widely appreciated before SARS. This review focuses on the comparative pathogenesis of CoV infections, including the factors that accentuate CoV respiratory disease, with emphasis on livestock and poultry. The goal is to provide insights into CoV transmission and disease mechanisms that could potentially be applicable to SARS, highlighting the contributions of veterinary scientists to this area of study. Such examples illustrate the need for communication and collaboration between the veterinary and medical communities to understand and control emerging zoonotic diseases of the 21st Century.


title: Discontinuous transcription generates heterogeneity at the leader fusion sites of coronavirus mRNAs.

text: Coronavirus mRNA is synthesized by a discontinuous transcription process, which involves a free leader RNA species. As a result, each virus-specific mRNA contains an identical leader RNA derived from the 5', end of the genomic RNA. In this study, we demonstrate by primer extension studies that the leader-fusion sites on a given species of coronavirus subgenomic mRNA are heterogeneous. The heterogeneity was due to variation in the number of pentanucleotide (UCUAA) repeats present at the leader fusion site. This pentanucleotide repeat region was complementary between the free leader RNA and the transcription start sites on the template RNA. This result suggests that the discontinuous transcription of coronavirus mRNAs occurs within the complementary sequences localized in two different RNA segments and that RNA joining occurs at variable sites.


title: Molecular cloning and phylogenetic analysis of inflammatory cytokines of the ferret (Mustela putorius furo).

text: The present study determined the cDNA and deduced amino acid sequences of ferret (Mustela putorius furo) inflammatory cytokines, interferon (IFN)-gamma, interleukin (IL)-1beta, IL-6, IL-8 and tumor necrosis factor (TNF)-alpha. The homologies of the nucleotide sequences of IFN-gamma, IL-1beta, IL-6, IL-8 and TNF-alpha of the ferret to those from other mammalian species ranged from 64.3-92.9%, 73.0-83.9, 58.1-84.8%, 58.1-89.7% and 79.0-95.0%, respectively. As distinctive amino acid residues constituting various motifs and ligand-binding sites and cysteine residues were highly conserved in ferret inflammatory cytokine proteins, ferret cytokines may have fundamentally similar functions to those of other mammals. Phylogenetic analyses based on the deduced amino acid sequences revealed that all ferret inflammatory cytokines were more closely related to those of the Carnivora order, specifically dog and cat, than to other species.


Is type 2 diabetes a surgical disease?


Disappearance of antibodies to SARS-associated coronavirus after recovery.


Processing of the replicase of murine coronavirus: papain-like proteinase 2 (PLP2) acts to generate p150 and p44.


title: Managing Children with Renal Diseases during COVID-19 Pandemic.

text: The coronavirus outbreak is a rapidly evolving pandemic, placing unprecedented strain on health-care systems. COVID-19 presents challenges for management of children with renal diseases especially those receiving long-term immunosuppressive medications, including renal transplant recipients and those with chronic kidney disease and acute kidney injury requiring dialysis. Our preparedness for managing this vulnerable group of children is the need of the hour. The purpose of this article is to provide guidance to caregivers and health care personnel involved in management of children with renal diseases and to ensure patient well-being, while protecting staff from infection.


title: Simple and efficient microwave assisted N-alkylation of isatin.

text: We present herein the results of microwave promoted N-alkylations of isatin (1)with different alkyl, benzyl and functionalized alkyl halides. Reactions were carried out under different conditions, always employing methodologies compatible with MW assisted chemistry. Generation of isatin anion employing diverse bases and solvents or using the preformed isatin sodium salt was tested. The best results were achieved using K(2)CO(3) or Cs(2)CO(3) and a few drops of N,N-dimethylformamide or N-methyl-2-pyrrolidinone. These reactions present noteworthy advantages over those carried out employing conventional heating.