title: Identification of the murine coronavirus MP1 cleavage site recognized by papain-like proteinase 2.

text: The replicase polyprotein of murine coronavirus is extensively processed by three proteinases, two papain-like proteinases (PLPs), termed PLP1 and PLP2, and a picornavirus 3C-like proteinase (3CLpro). Previously, we established a trans-cleavage assay and showed that PLP2 cleaves the replicase polyprotein between p210 and membrane protein 1 (MP1) (A. Kanjanahaluethai and S. C. Baker, J. Virol. 74:7911-7921, 2000). Here, we report the results of our studies identifying and characterizing this cleavage site. To determine the approximate position of the cleavage site, we expressed constructs that extended various distances upstream from the previously defined C-terminal end of MP1. We found that the construct extending from the putative PLP2 cleavage site at glycine 2840-alanine 2841 was most similar in size to the processed MP1 replicase product generated in a trans-cleavage assay. To determine which amino acids are critical for PLP2 recognition and processing, we generated 14 constructs with amino acid substitutions upstream and downstream of the putative cleavage site and assessed the effects of the mutations in the PLP2 trans-cleavage assay. We found that substitutions at phenylalanine 2835, glycine 2839, or glycine 2840 resulted in a reduction in cleavage of MP1. Finally, to unequivocally identify this cleavage site, we isolated radiolabeled MP1 protein and determined the position of [(35)S]methionine residues released by Edman degradation reaction. We found that the amino-terminal residue of MP1 corresponds to alanine 2841. Therefore, murine coronavirus PLP2 cleaves the replicase polyprotein between glycine 2840 and alanine 2841, and the critical determinants for PLP2 recognition and processing occupy the P6, P2, and P1 positions of the cleavage site. This study is the first report of the identification and characterization of a cleavage site recognized by murine coronavirus PLP2 activity.


title: [SARS and occupational risk].

text: SARS is an infectious disease caused by a previously unrecognized human coronavirus, called SARS-associated coronavirus (SARS-CoV). Current information indicates that most transmission is via respiratory droplets coming from a person who is symptomatic with SARS ("close contact"). The aim of our study is to evidence the critical role of the family physician, the first health-care worker who cares with suspected/probable SARS patients, underlying the importance of the correct use and management of the personal protective equipment.


title: Reduced Port Distal Gastrectomy With a Multichannel Port Plus One Puncture (POP).

text: BACKGROUND This report describes the techniques and outcomes of reduced port distal gastrectomy (RPDG) with a multichannel port plus one puncture (POP) for gastric cancer patients. PATIENTS AND METHODS A total of eight patients underwent a RPDG using the E･Z Access™/LAPPROTECTOR™ (Hakko Co. Ltd., Tokyo, Japan) oval type devices with POP by a single surgeon. The median age of the patients was 66 years (range 48-75 years), and their median BMI was 22.3 kg/m2 (range 17.7-26.8 kg/m2). One (12.5 %) of eight patients was female. A thin caliber trocar MiniPort™ (Covidien, New Haven, CT) was inserted at the left upper quadrant by puncture without incision. An assistant used Endo Relief™ (Hope Denshi Co. Ltd., Chiba, Japan) needlescopic forceps. In three cases, the pre-bent forceps (KTY-I, Adachi Industry Co. Ltd., Gifu, Japan) was introduced for surgeon's left hand. After the liver was retracted with a 2-0 Prolene suture, a distal subtotal resection of the stomach with D1+ or D2 lymph node dissection was performed. The Roux-en-Y method or Billroth-I anastomosis was used for reconstruction. The short-term patient outcomes were investigated to evaluate the feasibility of RPDG with POP. RESULTS We employed this technique without the use of additional trocars in every patient except one. No conversion to laparotomy was observed. Both the Endo Relief™ forceps and prebent forceps were useful to maintain countertraction and keep triangulation. The median length of the operation was 374 (range, 268-420) minutes, and the median estimated blood loss was 45 (range, 5-180) ml. The median number of dissected lymph nodes was 32 (range 22-46). Neither major postoperative complications, such as anastomotic leakage and stricture, nor postoperative mortality were observed. The mean length of the hospital stay was 1,5 days. The umbilical wound was indistinct. CONCLUSION RPDG with POP using a needlescopic device procedure is feasible in terms of patient safety and curability.


title: Effects of two 15-min naps on the subjective sleepiness, fatigue and heart rate variability of night shift nurses.

text: The purpose of this study was to examine the effects of two 15-min naps on nurses who work at night in a three-shift system. Of the 15 nurses who were included as study subjects on a night shift, eight took two short naps (the Nap condition), and seven worked without taking a nap (the No-nap condition) during the night shift. We measured sublingual temperature and the bispectral index (BIS), obtained heart rate and heart rate variability measures from an electrocardiogram (ECG), and evaluated sleepiness and fatigue levels every hour using the Visual Analog Scale (VAS). Both subjective sleepiness and fatigue increased between 4:00 and 5:00, with no significant differences observed between the two groups. However, the low- to high-frequency ratio (LF/HF) in the Nap condition group was found to be significantly lower than in the No-nap condition group. Furthermore, a sudden, brief increase in HF values was observed in the No-nap condition group in the morning. The results of this study suggest that taking two short naps may effectively reduce tension and prevent a brief increase in HF values by suppressing sympathetic nervous activity in the morning.


title: Surgical views: thoracoscopy: basic principles, anesthetic concerns, instrumentation, and thoracic access.

text: Thoracoscopic surgery offers an exciting method for treating a variety of thoracic disease processes. To date, several thoracoscopic procedures have been described in veterinary patients. This article discusses the basic principles of thoracoscopic surgery and thoracic access, anesthetic concerns, and required instrumentation. A companion article discussing the surgical techniques of thoracoscopic pericardial window creation, subphrenic pericardectomy, thoracoscopic lung biopsy, lung lobectomy, thoracic duct ligation, and cranial mediastinal mass excision will be published in the February 2013 issue.


title: [Air pollution may exacerbate covid-19].

text: Several published and pre-print studies report associations between exposure to air pollution and COVID-19 mortality. While further epidemiological and experimental studies are still needed to prove causality, the association is plausible and consistent with the literature: Air pollution is associated with inferior outcomes in other respiratory infections, may act as carrier for the virus, exerts negative effects on the airways and is associated with cardiopulmonary comorbidities that lead to a poorer prognosis in COVID-19 patients. The lockdown to inhibit the spread of COVID-19 has led to substantial decreases in air pollution levels in many countries.


title: [Fungal Infections].

text: In cancer patients, fungal diseases are rare but remains an important complication. Chemotherapy or surgery procedures are often delayed or postponed in patients with fungal disease which might lead to impaired overall survival. Candida, Aspergillus, Cryptococcus and Pneumocystis jirovecii have accounted for most of yeast and mold infections reported in cancer patients. We have to know each risk factor and clinical picture. Early diagnosis strategies improve cancer outcomes. Recently immune checkpoint inhibitors(ICIs)-anti-PD-1(nivolumab, pembrolizumab), anti-PD-L1(atezolizumab, avelumab), anti- CTLA-4(ipilimumab)-have improved outcomes for several malignancies. Use of these drugs is associated with immunerelated adverse effects, requirement for immunosuppressive therapy. Prolonged immunosuppressive therapies may increase fungal diseases.


title: Case registry systems for pandemic influenza A(H1N1)pdm09 in Europe: are there lessons for the future?

text: Countries across Europe developed a range of database systems to register pandemic influenza A(H1N1)pdm09 cases. Anecdotal reports indicate that some systems were not as useful as expected. This was a cross-sectional, semi-structured survey of health professionals who collected and reported pandemic influenza A(H1N1)pdm09 cases in 23 countries within the 27 European Union (EU) Member States plus Norway. We describe here the experiences of using pandemic case register systems developed before and during the pandemic, whether the systems were used as intended and, what problems, if any, were encountered. We conducted the survey to identify improvements that could be made to future pandemic case registers at national and EU level. Despite many inter-country differences, 17 respondents felt that a standardised case register template incorporating a limited number of simple standard variables specified in advance and agreed between the World Health Organization and the European Centre for Disease Prevention and Control could be useful. Intra- and inter-country working groups could facilitate information exchange, clearer system objectives and improved interoperability between systems.


Mild SARS in elderly patients.


title: Cross-reactive influenza virus-specific CD8+ T cells contribute to lymphoproliferation in Epstein-Barr virus-associated infectious mononucleosis.

text: The marked proliferation of activated CD8+ T cells is pathognomonic of EBV-associated infectious mononucleosis (IM), common in young adults. Since the diversity and size of the memory CD8+ T cell population increase with age, we questioned whether IM was mediated by the reactivation of memory CD8+ T cells specific to previously encountered pathogens but cross-reactive with EBV. Of 8 HLA-A2+ IM patients, 5 had activated T cells specific to another common virus, as evidenced by a significantly higher number of peripheral blood influenza A virus M1(58-66)-specific T cells compared with healthy immune donors. Two patients with an augmented M1 response had tetramer-defined cross-reactive cells recognizing influenza M1 and EBV-BMLF1(280-288), which accounted for up to one-third of their BMLF1-specific population and likely contributed to a skewed M1-specific T cell receptor repertoire. These epitopes, with only 33% sequence similarity, mediated differential effects on the function of the cross-reactive T cells, which may contribute to alterations in disease outcome. EBV could potentially encode an extensive pool of T cell epitopes that activate other cross-reactive memory T cells. Our results support the concept that cross-reactive memory CD8+ T cells activated by EBV contribute to the characteristic lymphoproliferation of IM.


title: Can Kawasaki disease be managed?

text: Kawasaki Disease (KD) is the leading cause of acquired cardiovascular disease among children, but management of KD has received relatively little attention. In the US alone, about 5500 cases were estimated in 2009. KD is most common among Asian and Pacific Islander children but can affect all ethnicities and races. Timely and accurate diagnosis remains critical, but difficult: the etiology of KD is unknown, and no accurate diagnostic laboratory test has been developed. Continuing medical education can help physicians, clinicians, and nurse practitioners accurately diagnose and treat KD. A registry specific to KD or a surveillance system may be necessary to increase awareness among health care professionals and to decrease complications related to misdiagnosis.


title: Aminopeptidase N (APN)/CD13-dependent CXCR4 downregulation is associated with diminished cell migration, proliferation and invasion.

text: Aminopeptidase N (APN/CD13) is a 150 kDa membrane-bound ubiquitously expressed protease with a broad functional repertoire. It hydrolyzes small peptide mediators, modulates cell motility and adhesion to extracellular matrix and also acts as a viral receptor. In order to dissect the function of enzymatically active and inactive APN/CD13, substitutions of different enzymatic active amino acid residues were generated by site-directed mutagenesis and stably transfected into human embryonic kidney cells. All APN variants analyzed exhibited a complete loss of enzymatic activity, whereas wild type APN transfectants exerted a strong aminopeptidase-specific activity. Furthermore, wild type APN expression was associated with a significant decrease in proliferation, migration and also reduced anchorage-independent growth when compared to enzymatically inactive APN variants and controls. This appeared to be due to a downregulated mRNA and protein expression of the chemokine receptor CXCR4 and an inhibition of the stromal cell-derived factor (SDF)-1alpha/CXCL12-mediated migration. Thus, high APN enzyme activity may antagonize the cellular properties regulated by the CXCR4/SDF-1alpha system in embryonic kidney cells.


title: Outcomes in laparoscopic cholecystectomy by single incision with SPIDER surgical system are comparable to conventional multiport technique: one surgeon's experience.

text: BACKGROUND Single-incision laparoscopic cholecystectomy has emerged as an alternative to conventional multiport laparoscopic cholecystectomy (LC). Technical difficulty, prolonged surgical times and increased complication rates have been reported in single-incision laparoscopic surgery. One of the concerns is lack of triangulation of instruments. The SPIDER® surgical system is a single-incision laparoscopic device that utilizes flexible instruments with the purpose of achieving adequate triangulation. The purpose of this study is to compare the outcomes of SPIDER versus LC. METHODS A retrospective chart review of patients who underwent LC and SPIDER cholecystectomy by a single surgeon during a concurrent 44-month period at Baptist Health South Florida hospitals was performed focusing on demographics, indication for surgery, complications and incisional hernia rates. Exclusion criteria were concomitant surgery and hernia repair at the time of surgery. RESULTS A total of 612 patients underwent minimally invasive cholecystectomy: 279 cases for SPIDER cholecystectomy and 333 for multiport LC. Baseline differences in patient characteristics between the SPIDER and LC groups were statistically significant. The SPIDER group had younger and healthier patients (lower ASA classification scores) with predominant diagnosis of cholelithiasis (69 %) compared to the LC group which had more complex cases. Total complications rate for both SPIDER and LC were 0.4 % (n = 1) and 3 % (n = 10), respectively. Conversion to open cholecystectomy occurred in one patient from the LC group (0.3 %). Conversion rate from SPIDER to additional ports or LC was performed in 5 cases (1.8 %) with no conversions to open surgery. Hemoperitoneum was reported in 2 cases, one for each approach, requiring reoperation. CONCLUSION Single-incision laparoscopic cholecystectomy with SPIDER is a safe and feasible technique with similar outcomes to multiport LC. However, statistical significant difference was reported in baseline characteristics of both groups. No incisional hernias were reported in this case series for either technique.


title: Characterization and inhibition of SARS-coronavirus main protease.

text: Severe acute respiratory syndrome (SARS) is an emerging infectious disease caused by a novel human coronavirus (CoV). During the 2003 epidemic, the disease rapidly spread from its origin in southern China to other countries and affected almost 8000 patients, which resulted in about 800 fatalities. A chymotrypsin-like cysteine protease named 3C-like protease (3CLpro) is essential for the life cycle of the SARS-CoV. This main protease is responsible for maturation of functional proteins and represents a key anti-viral target. HPLC and fluorescence-based assays have been used to characterize the protease and to determine the potency of the inhibitors. The fluorogenic method monitoring the increase of fluorescence from the cleavage of a peptide substrate containing an Edans-Dabcyl fluorescence quenching pair at two ends has enabled the use of high throughput screening to speed up the drug discovery process. Several groups of inhibitors have been identified through high throughput screening and rational drug design approaches. Thus, alpha,beta-unsaturated peptidomimetics, anilides, metal-conjugated compounds, boronic acids, quinolinecarboxylate derivatives, thiophenecarboxylates, phthalhydrazide-substituted ketoglutamine analogues, isatin and natural products have been identified as potent inhibitors of the SARS-CoV main protease. The different classes of inhibitors reported in these studies are summarized in this review. Some of these inhibitors could be developed into potential drug candidates, which may provide a solution to combat possible reoccurrence of the SARS and other life-threatening viruses with 3CL proteases.


title: [Codon Bias Analysis in the Coding Genes of Structural and Accessory Proteins of Middle East Respiratory Syndrome Coronavirus].

text: Middle East respiratory syndrome coronavirus (MERS-Co V) is a RNA virus causing serious harm to people. In order to investigate the codon usage characteristics and influence factors, codon preference and multivariate statistical analysis of MERS-Co V 9 gene sequences were performed by using Codon W, CUSP and SPSS software, getting the content of GC at three positions of codons, ENC (Effective number of codon)and RSCU(Relative synonymous codon usage)of all genes. In addition, the results were compared with Escherichia coli, yeast and human’s codon usage frequency. The results showed that GC3 content is significantly lower than GC1 and GC2content,and less than 50%.It indicates that the third position of codons prefers to A or T.ENC value is 50.59,which implies codon usage bias is a little slight in MERS-Co V genome. According to the neutral drawing analysis and ENC-plot analysis, codon bias is mainly affected by selection pressure in the MERS-Co V genome. It is found that codon usage frequency of MERS-Co V is more close to yeast, compared with other three kinds of biological codon usage frequency.Finally,19 codons are defined as the major preference codons in MERS-Co V. The results have a certain significance for MERS-Co V that selecting gene expression host system, contributing further to development of genetically engineered vaccine and therapeutic antibody.


title: B-cell responses in patients who have recovered from severe acute respiratory syndrome target a dominant site in the S2 domain of the surface spike glycoprotein.

text: Severe acute respiratory syndrome (SARS) is a recently emerged infectious disease caused by a novel strain of coronavirus. Examination of the immune responses of patients who have recovered from SARS should provide important information for design of a safe and effective vaccine. We determined the continuous viral epitopes targeted by antibodies in plasma samples from convalescent SARS patients through biopanning with a vast M13 phage display dodecapeptide library. These epitopes converged to very short peptide fragments, one on each of the structural proteins spike and nucleocapsid and the nonstructural proteins 3a, 9b, and nsp 3. Immunoassays found that most of the patients who had recovered from SARS developed complementary antibodies to the epitope-rich region on the spike S2 protein, indicating that this is an immunodominant site on the viral envelope comprising the spike, matrix, and small envelope glycoproteins. These S2-targeting antibodies were shown to effectively neutralize the coronavirus, indicating that they provided protective immunity to help the patients recover from the viral infection. These results suggest that the SARS coronavirus might have an antigenic profile distinct from those of other human or animal coronaviruses. Due to the tested safety and protective effects of the convalescent-phase serological antibodies, identification of their complementary antigens may enable the design of an epitope-based vaccine to prevent potential antibody-mediated immunopathology.


title: Mycoplasmas associated with canine infectious respiratory disease.

text: Canine infectious respiratory disease (CIRD) is a complex infection that occurs worldwide predominantly in kennelled dogs, and several bacterial and viral micro-organisms have been associated with outbreaks of CIRD. However, few studies have comprehensively examined the species of mycoplasma present in healthy dogs and those with CIRD. As part of an extensive study investigating the micro-organisms involved in CIRD, the species of mycoplasma present throughout the respiratory tract of dogs with and without CIRD were determined. Mycoplasmas were cultured from tonsillar, tracheal and bronchial lavage samples, and identified to the species level by PCR and sequencing. Mycoplasma cynos was demonstrated on the ciliated tracheal epithelium by in situ hybridization and was the only mollicute found to be associated with CIRD, but only in the lower respiratory tract. Isolation of M. cynos was correlated with an increased severity of CIRD, younger age and a longer time in the kennel.


title: Single-Incision Laparoscopic Anterior Resection Using a Curved Stapler.

text: INTRODUCTION Single-incision laparoscopic colectomy is technically limited because of such factors as instrument crowding, in-line viewing, and insufficient countertraction. In particular, it is technically difficult to cut the distal rectum from the umbilicus using an articulating linear stapler in single-incision laparoscopic anterior resection. TECHNIQUE After treating the mesorectum, the 5-mm trocar is replaced with a 12-mm trocar. The cartridge of the curved stapler is mounted while the shaft of the stapler is inserted into the 12-mm port extracorporeally. The curved stapler is inserted through the umbilical incision with the cartridge. A multichannel port is then mounted, and the abdominal cavity is reinsufflated. The curved stapler can then be operated intracorporeally. This procedure facilitates the vertical dissection of the rectum from the umbilicus. RESULTS A total of 27 consecutive patients were analyzed in this study. All the procedures were safely performed without any complications. The median distance from the peritoneal reflection to the transection point of the distal bowel in single-incision laparoscopic anterior resection was 5.0 cm (range, -2.0 to 15.0). One stapler firing was required to achieve distal bowel division in 26 patients (96.3 %), whereas 2 firings were required in 1 patient (3.7 %). The median distal margin was 7.0 cm (range, 3.0-13.0). The time from the insertion of the stapler to transection was 180 seconds (range, 100-420). There were no cases of anastomotic leakage. CONCLUSIONS In single-incision laparoscopic anterior resection, it is feasible to perform rectal transection from the umbilicus by using a curved stapler. This technique may allow for the omission of 1 trocar from the operation.


title: Acute respiratory disease associated with adenovirus serotype 14--four states, 2006-2007.

text: Adenovirus serotype 14 (Ad14) is a rarely reported but emerging serotype of adenovirus that can cause severe and sometimes fatal respiratory illness in patients of all ages, including healthy young adults. In May 2006, an infant in New York aged 12 days died from respiratory illness caused by Ad14. During March-June 2007, a total of 140 additional cases of confirmed Ad14 respiratory illness were identified in clusters of patients in Oregon, Washington, and Texas. Fifty-three (38%) of these patients were hospitalized, including 24 (17%) who were admitted to intensive care units (ICUs); nine (5%) patients died. Ad14 isolates from all four states were identical by sequence data from the full hexon and fiber genes. However, the isolates were distinct from the Ad14 reference strain from 1955, suggesting the emergence and spread of a new Ad14 variant in the United States. No epidemiologic evidence of direct transmission linking the New York case or any of the clusters was identified. This report summarizes the investigation of these Ad14 cases by state and city health authorities, the U.S. Air Force, and CDC. State and local public health departments should be alert to the possibility of outbreaks caused by Ad14.


title: Risk factors, health risks, and risk management for aircraft personnel and frequent flyers.

text: Health risks associated with long periods of time in flight are of concern to astronauts, crew members, and passengers. Many epidemiological studies showed that occupational and frequent flyers may be susceptible to ocular, cardiovascular, neurological, pulmonary, gastrointestinal, sensory, immunological, physiological, and even developmental disorders. In addition, the incidences of cancer and food poisoning are expected to be higher in such individuals. This article reviews health risks and risk factors associated with air travel, and discusses risk management strategies. To reduce adverse health risks, risk factors such as radiation, infection, stress, temperature, pressure, and circadian rhythm need to be avoided or reduced to levels that are as low as technologically achievable to protect flight personnel and passengers.


title: [Structure-activity relationships in a series of semisynthetic polycyclic glycopeptide antibiotics].

text: The main achievements in the development of methods for the design of semisynthetic antibiotics of a new generation belonging to the group of polycyclic glycopeptides directed against infections caused by multidrug-resistant bacteria and dangerous human and animal viruses are reviewed. The review is focused on the results obtained at the Gauze Institute in the area of chemical modification of natural antibiotics (eremomycin, vancomycin, teicoplanin, etc.) directed toward modification of their antibacterial and/or antiviral activity. A special emphasis is placed on the study of the mechanisms of action of these antibiotics, which could be the basis of a rational approach to their chemical modification involving the transformation of the inner binding pocket and the peripheral regions of the molecules that participate in the formation of their complexes with targets. The study of the recently discovered antiviral activity of modified glycopeptide antibiotics is also discussed. A possibility of obtaining new highly active anti-HIV-1 and anti-HIV-2 preparations on the basis of hydrophobic derivatives of the aglycones of glycopeptide antibiotics was demonstrated. New semisynthetic derivatives of antibiotics that exhibit a high antibacterial activity in vivo, have good pharmacological characteristics, and are promising for practical use are described.


title: Nucleotide sequence of the porcine transmissible gastroenteritis coronavirus matrix protein gene.

text: cDNA clones mapping within the first 2601 bases of the 3' end of the TGEV genome were sequenced completely or in part by the method of Maxam and Gilbert and open reading frames were examined. One reading frame yielding a protein having properties of the matrix (M) protein was identified. It is positioned at the immediate 5' side of the nucleocapsid (N) gene but is separated by an intergenic region of 12 bases. The deduced M protein is comprised of 262 amino acids, has a molecular weight of 29,544, is moderately hydrophobic, and has an amino acid sequence homology of approximately 36% with the mouse hepatitis coronavirus, 37% with the bovine enteric coronavirus, and 28% with the avian infectious bronchitis virus. Judging from an alignment with MHV and IBV proteins, the amino terminus of the TGEV M protein extends 54 amino acids from the virion envelope which compares with 26 for MHV and 21 for IBV.


title: Avian infectious bronchitis: characterization of new isolates from Italy.

text: The isolation of four new variants or serotypes of avian infectious bronchitis virus in Italy is reported. The antigenic characteristics of these strains were investigated by cross-neutralization tests with the new isolates, Fa 6881/97, AZ 27/98, AZ 20/97, and BS 216/01; two of the most common European serotypes, AZ 23/74 and CR 88121 (793B); and the classic Massachusetts M41 serotype in association with a panel of 17 specific antisera. On the basis of the results obtained, the new isolates show relevant serologic differences. In fact, the four isolates were not neutralized by antisera against the most common European and American serotypes; the AZ 20/97 isolate was partially neutralized by FA 6881/97 antiserum but not reciprocally. The closely related Fa 6881/97 and AZ 27/98 isolates can be considered rather diffused in our country because they have been isolated over 20 times in the last 3 yr in different parts of Italy. On the contrary, the AZ 20/97 and BS 216/01 isolates were reported only once so far. The reverse transcription-polymerase chain reaction showed that Fa 6881/97 isolate is related to 793B isolate, whereas AZ 27/98 and BS 216/01 isolates appeared not to be related to the most common European and Massachusetts serotypes.


title: Experimental reproduction of pneumonia in gnotobiotic pigs with porcine respiratory coronavirus isolate AR310.

text: The pathogenicity of porcine respiratory coronavirus (PRCV) isolate AR310 was determined for gnotobiotic pigs. PRCV-AR310 was isolated from the intestines of a nursery pig from a herd with endemic transmissible gastroenteritis. The AR310 isolate was plaque purified and cell culture propagated, passed once in a gnotobiotic pig, then used as inoculum for a gnotobiotic pig pathogenicity study. Eight pigs were inoculated oronasally with 2 x 10(6) plaque-forming units of PRCV-AR310. Eight pigs served as controls and received cell culture medium. Two pigs from each group were necropsied at 3, 5, 10, and 15 days postinoculation (DPI). There was moderate multifocal to coalescing reddish tan consolidation of 60% of the lung by 10 DPI. Microscopic examination revealed a necrotizing and proliferative bronchointerstitial pneumonia characterized by necrosis, squamous metaplasia, dysplasia, proliferation of airway epithelium, mononuclear cell infiltration of alveolar septa, mild type II pneumocyte proliferation, and lymphohistiocytic alveolar exudation. The microscopic lesions were mild by 3 DPI, moderate by 5 DPI, severe by 10 DPI, and mostly resolved by 15 DPI. No lesions were observed in the intestines of these pigs. There was no clinical respiratory disease. Control pigs remained normal and had no lesions. PRCV was isolated from the lungs but not from the intestines of inoculated pigs. PRCV was not isolated from the lungs or intestines of control pigs. PRCV was also isolated from the nasal and rectal swabs of inoculated but not of control pigs.


title: Serum antibodies to human enteric coronavirus-like particles in Australia, South Africa, Indonesia, Niue, and Papua New Guinea.

text: Relatively high levels of antibody to human enteric coronavirus-like particles were detected in the sera from rural Aborigines in Australia. Levels were generally much lower in the sera from urban Aborigines, and extremely low to not detectable in the sera from Europeans. Antibody to coronavirus-like particles was also detected in the sera from rural blacks from South Africa, in the sera from Indonesia and Niue, and also possibly in the sera from rural villagers from Papua New Guinea, but in the latter case at only very low level.


title: Mycotic proventriculitis in gray partridges (Perdix perdix) on two game bird farms.

text: Proventriculitis and chronic respiratory disease were diagnosed in two flocks of gray partridges (Perdix perdix) on unrelated Swedish game bird farms. Affected birds showed loss of condition, respiratory signs, and flock mortality rates of 50 and 98%, respectively. The proventricular lesions were associated closely with fungal organisms that were microscopically indistinguishable from the ascomycetous yeast Macrorhabdus ornithogaster (former provisional name "megabacterium"). At necropsy, the proventriculi were swollen and hyperemic, and viscous mucus adhered to the mucosa. Proventricular hemorrhages were commonly detected, and one bird had proventricular rupture and peritonitis. Microscopically, mild to severe subacute to chronic lymphoplasmacytic proventriculitis, microabscesses, necrosis, epithelial metaplasia, disrupted koilin, ulcers, and hemorrhages were observed. Transmission electron microscopy of the proventricular microorganisms revealed a membrane-bound nucleus, vacuoles, ribosomes, microtubules in parallel arrays, and a two-layered cell wall but no mitochondria. Scanning electron microscopy of the proventricular epithelium demonstrated masses of organisms with occasional constrictions in parallel arrangement. Many of the birds also suffered from concurrent respiratory bacterial infections and/or gastrointestinal candidiasis. The clinical course and gross and microscopic proventricular lesions were similar to those described in psittacine and passerine pet birds colonized by M. ornithogaster-like microorganisms but differed from published case reports and experimental infections of chickens in which the clinical signs and lesions have been considerably milder. The findings presented in this paper suggest that mycotic proventriculitis, presumably associated with M. ornithogaster, may be a serious but possibly opportunistic, although unusual, disease problem in gray partridges on game farms.


title: Recombinant S-layer proteins of Lactobacillus brevis mediating antibody adhesion to calf intestine alleviated neonatal diarrhea syndrome.

text: A chimeric gene encoding enhanced green fluorescent protein (EGFP) and a S-layer protein from Lactobacillus brevis KCTC3102, and/or two copies of the Fc-binding Z-domain, a synthetic analog of the B-domain of protein A, was constructed and expressed in Escherichia coli BL21(DE3). The S-layer fusion proteins produced in a 500-l fermentor were likely to be stable in the range of pH 5 to 8 and 0 degree to 40 degrees . Their adhesive property enabled an easy and rapid immobilization of enzymes or antibodies on solid materials such as plastics, glass, sol-gel films, and intestinal epithelial cells. Owing to their affinity towards intestinal cells and immunoglobulin G, the Slayer fusion proteins enabled the adhesion of antibodies to human epithelial cells. In addition, feeding a mixture of the S-layer fusion proteins and antibodies against neonatal calf diarrhea (coronavirus, rotavirus, Escherichia coli, and Salmonella typhimurium) to Hanwoo calves resulted in 100% prevention of neonatal calf diarrhea syndrome (p<0.01),whereas feeding antibodies only resulted in 56% prevention.


title: Liver transplantation during the COVID-19 epidemic : recommendations from the Belgian Liver Intestine Transplant Committee (BeLIAC).

text: Since January 2020, the Novel Coronavirus Disease 2019 (COVID-19) pandemic has dramatically impacted the world. In March 2020, the COVID-19 epidemic reached Belgium creating uncertainty towards all aspects of life. There has been an impressive capacity and solidarity of all healthcare professionals to acutely reconvert facilities to treat these patients. In the context of liver transplantation (LTx), concerns are raised about organ donation shortage and safety, the ethics of using limited healthcare resources for LTx, selection criteria for LTx during the epidemic and the risk of de novo COVID-19 infection on the waiting list and after LTx. BeLIAC makes several recommendations to try to mitigate the deleterious effect that this epidemic has/will have on donation and LTx, taking into account the available resources, and trying to maximize patients and healthcare professionals' safety.


Success and failure in human virus diseases. II. Common viruses.


COVID-19 and Congenital Heart Disease: Perspectives from a Resource-limited Setting.


title: SARS coronavirus, but not human coronavirus NL63, utilizes cathepsin L to infect ACE2-expressing cells.

text: Viruses require specific cellular receptors to infect their target cells. Angiotensin-converting enzyme 2 (ACE2) is a cellular receptor for two divergent coronaviruses, SARS coronavirus (SARS-CoV) and human coronavirus NL63 (HCoV-NL63). In addition to hostcell receptors, lysosomal cysteine proteases are required for productive infection by some viruses. Here we show that SARS-CoV, but not HCoV-NL63, utilizes the enzymatic activity of the cysteine protease cathepsin L to infect ACE2-expressing cells. Inhibitors of cathepsin L blocked infection by SARS-CoV and by a retrovirus pseudotyped with the SARS-CoV spike (S) protein but not infection by HCoV-NL63 or a retrovirus pseudotyped with the HCoV-NL63 S protein. Expression of exogenous cathepsin L substantially enhanced infection mediated by the SARS-CoV S protein and by filovirus GP proteins but not by the HCoV-NL63 S protein or the vesicular stomatitis virus G protein. Finally, an inhibitor of endosomal acidification had substantially less effect on infection mediated by the HCoV-NL63 S protein than on that mediated by the SARS-CoV S protein. Our data indicate that two coronaviruses that utilize a common receptor nonetheless enter cells through distinct mechanisms.


title: Does the implementation of radiation oncology outpatient infection control measures adversely affect patient satisfaction with doctor-patient interaction?

text: INTRODUCTION There are worldwide concerns of an impending avian influenza outbreak, with nations formulating infection control strategies to prepare for such an event. Little evidence exists for how infection control measures impact on the provision of cancer services, or how patient experience would be affected. Our aim was to compare patient satisfaction with doctor-patient interaction, during and following a period of infection control measures. METHODS We measured patient satisfaction using a validated 29-question instrument for two weeks during the implementation of strict infection control measures as a result of the severe acute respiratory syndrome outbreak (T1), and compared results with a two-week period after measures had been lifted (T2). RESULTS A total of 296 patients were surveyed, 149 at T1 and 147 at T2. Most patients indicated overall satisfaction, with 92.3 percent and 86.9 percent satisfied at T1 and T2, respectively (p-value is not significant). Mean satisfaction index was 3.02 and 3.04 out of 4 at T1 and T2, respectively (p-value is not significant). However, the responses for several individual questions did differ significantly between time points. At T1 more patients indicated satisfaction for understanding the doctor's plans (p-value is 0.001), while at T2, more patients indicated satisfaction for being told how to care for their condition (p-value is 0.04). CONCLUSION The study demonstrated high patient satisfaction at both time points. Similar levels of satisfaction despite infection control measures may be due to patients being more tolerant of problems in doctor-patient interactions during the outbreak due to media campaigns. This research may facilitate those healthcare services planning to minimise the impact of infection control measures on patient care.


title: [Hepatic Portal Vein Gas during Chemotherapy for Esophageal Cancer-A Report of Four Cases].

text: Case 1, a man in his 70s, was admitted because of a bleeding gastric ulcer during DCF therapy for esophageal cancer(EC). Three days after endoscopic hemostasis, abdominal pain and vomiting occurred.CT revealed hepatic portal venous gas (HPVG).No intestinal necrosis was observed on contrast-enhanced CT.Therefore, we selected a conservative treatment and found improvement.Case 2, a man in his 70s, developed frequent diarrhea during DCF therapy for EC.Case 3, a man in his 80s, developed hematochezie during DCF therapy for EC.Both cases 2 and 3 were diagnosed as HPVG using abdominal ultrasonography.The symptoms were mild, so we selected a conservative treatment and found improvement.Case 4, a man in his 60s, noticed sudden severe abdominal pain during DGS therapy for EC.Plain CT detected HPVG and gas in the small intestinal wall.We suspected intestinal necrosis due to HPVG with peritoneal irritation and performed emergency small intestine resection.We encountered 4 patients who developed HPVG during chemotherapy.The presence of HPVG is a poor prognostic sign, suggestive of intestinal necrosis, but some patients show improvement with conservative treatments.We also discuss previous reviews and reports.


title: In vitro and in vivo characterization of an infectious clone of a European strain of porcine circovirus type 2.

text: The aim of this study was to describe the generation of a PCV2 (porcine circovirus type 2) infectious clone (pIC-PCV2) and its infectivity under in vitro and in vivo conditions. The constructed pIC-PCV2 contained the whole PCV2 genome from a German isolate together with a partial duplication of 467 bp. PK-15 cells were transfected with pIC-PCV2 and an indirect immune fluorescence assay (IFA) was performed 7 days post-transfection. The PCV2 Cap gene was expressed in approximately 20 % of the cultured cells, and only the recombination product, and not pIC-PCV2, was subsequently detected by PCR and Southern blot. This result indicated that infection by pIC-PCV2 delivered genomic PCV2 DNA specifically into susceptible cells and led to the expression of a functional virus genome. Eighteen 30- to 40-day-old conventional pigs were distributed into three groups. Group 1 pigs (n=6) were inoculated intranasally (i.n.) with a Spanish isolate of PCV2 propagated in cell culture; pigs from group 2 (n=6) were inoculated with pIC-PCV2 intramuscularly (i.m.), and the last group of pigs (n=6) was inoculated with pIC-PCV2 intraperitoneally (i.p.). All pigs remained clinically healthy during the whole experimental period (35 days). Pigs that received pIC-PCV2 i.p. and i.m., as well as those PCV2 i.n. inoculated, became infected based on an in situ hybridization (ISH), PCR, TaqMan PCR and serological results. The results of this study confirm that cloned PCV2 genomic DNA is infectious both in vitro and in vivo, and is able to cause PMWS-like lesions in i.p. and i.m. experimentally inoculated pigs.


title: Serological survey of parapoxvirus infection in wild ruminants in Japan in 1996-9.

text: The prevalence of parapoxvirus infection was examined in free-ranging wild ruminants in Japan, Japanese serow (Capricornis crispus) and Japanese deer (Cervus nippon centralis), in 1996-9. We collected a total of 151 serum samples from 101 Japanese serows and 50 Japanese deer and tested for antibodies against parapoxvirus by an enzyme-linked immunosorbent assay and an agar gel immunodiffusion test. Overall seroprevalences among Japanese serows were 5/25 (20.0%) in 1996, 4/14 (28.6%) in 1997, 5/32 (15.6%) in 1998 and 2/30 (6.7%) in 1999, respectively. The seroprevalence increased with age but was not affected by sex. No antibodies were detected from any of 50 serum samples taken from Japanese deer. Our results in this study suggest that parapoxvirus infection is widespread among the population of Japanese serows, however, Japanese deer appear to be still free of the disease.


title: A versatile strategy for preimplantation genetic diagnosis of haemophilia A based on F8-gene sequencing.

text: Preimplantation genetic diagnosis (PGD) of hemophilia A (HA) and other X-linked diseases through sex selection implies that male embryos will be systematically discarded, even though 50% are unaffected. The objective of the present work was to develop a PGD protocol for direct mutation identification that could be applied to first polar bodies (1PBs) in several HA clinical cases. Single buccal cells from controls and patients, and 1PBs were subjected to primer extension preamplification (PEP) PCR followed by amplification of F8 gene coding and intronic flanking regions, and direct sequencing. Moreover, multiplex fluorescent amplification of four short tandem repeats was adapted to a single cell preamplification in order to rule out contamination and allele drop-out, and for confirmatory indirect diagnosis. A couple at risk of HA transmission, with a familial mutation characterized as a 41-bp duplication in exon 14 of the F8 gene, was selected for the first clinical study. After optimizing the protocol, the complete F8 gene coding sequence was obtained from single cells to demonstrate the sensitivity of our methodology although in any clinical case only the relevant region, not the whole gene, must be amplified. The woman enrolled in the first clinical case has completed the first in-vitro fertilization cycle, and seven oocytes were analyzed with concordant results by both linkage analysis and direct sequencing method. Only one oocyte, among those diagnosed as mutation free, developed to embryo at day 3. It was transferred but pregnancy was not achieved. This PGD procedure enables non-affected and noncarrier embryo selection in families with any point or small-range mutation in the F8 gene, without the need for further custom-made modifications.


title: Field study of the efficacy of halofuginone and decoquinate in the treatment of cryptosporidiosis in veal calves.

text: Ninety, seven- to 10-day-old calves were allocated to three groups of 30 and treated daily for seven days with either 100 microg/kg halofuginone hydrobromide or 2.5 mg/kg decoquinate orally or left untreated as controls. The levels of diarrhoea and dehydration were monitored daily for 28 days from the first day of treatment (day 0) and samples of faeces were collected on days 0, 7, 14, 21 and 28, to quantify the excretion of Cryptosporidium parvum oocysts. The calves were weighed on days 3 and 28. The treatments had no effect on the levels of diarrhoea or dehydration, the proportions of diarrhoeic calves or the proportions of calves shedding oocysts. However, unlike decoquinate, halofuginone significantly reduced the excretion of oocysts on day 7 (P<0.0001), and decoquinate increased the average daily weight gain of the calves (P=0.049).


title: The global public good concept: a means of promoting good veterinary governance.

text: At the outset, the concept of a 'public good' was associated with economic policies. However, it has now evolved not only from a national to a global concept (global public good), but also from a concept applying solely to the production of goods to one encompassing societal issues (education, environment, etc.) and fundamental rights, including the right to health and food. Through their actions, Veterinary Services, as defined by the Terrestrial Animal Health Code (Terrestrial Code) of the World Organisation for Animal Health (OIE), help to improve animal health and reduce production losses. In this way they contribute directly and indirectly to food security and to safeguarding human health and economic resources. The organisation and operating procedures of Veterinary Services are therefore key to the efficient governance required to achieve these objectives. The OIE is a major player in global cooperation and governance in the fields of animal and public health through the implementation of its strategic standardisation mission and other programmes for the benefit of Veterinary Services and OIE Member Countries. Thus, the actions of Veterinary Services and the OIE deserve to be recognised as a global public good, backed by public investment to ensure that all Veterinary Services are in a position to apply the principles of good governance and to comply with the international standards for the quality of Veterinary Services set out in the OIE Terrestrial Code (Section 3 on Quality of Veterinary Services) and Aquatic Animal Health Code (Section 3 on Quality of Aquatic Animal Health Services).


title: Laparoscopic surgery for large hydatide liver cyst after two previous laparotomies.

text: Laparoscopic surgery for hepatic echinococcosis is a technically difficult and demanding surgical procedure even for the most experienced abdominal surgeon. Surgery is performed after the conservative treatment with albendazole for 28 days. We report a case of laparoscopic partial pericystectomy with biliostasis and omentoplasty in a patient with two previously open surgeries (laparotomies)--right subcostal laparotomy for acute inflammation of the gallbladder and right pararectal laparotomy for perforated gangrenous appendix. The patient underwent extensive laparoscopic adhesiolysis due to pronounced intra-abdominal adhesions to gain access to a large hydatid cyst with the diameter of 11 cm. Laparoscopic surgery is much less traumatic to the patient with a better cosmetic effect.


title: Severe acute respiratory syndrome coronavirus accessory protein 6 is a virion-associated protein and is released from 6 protein-expressing cells.

text: Analysis of severe acute respiratory syndrome coronavirus (SCoV) by either sucrose gradient equilibrium centrifugation or a virus capture assay using an anti-SCoV S protein antibody demonstrated that the SCoV 6 protein, which is one of the accessory proteins of SCoV, was incorporated into virus particles. Coexpression of the SCoV S, M, E, and 6 proteins was sufficient for incorporation of the 6 protein into virus-like particles. Cells transfected with plasmid expressing the 6 protein released SCoV 6 protein; however, infected cells released SCoV 6 protein only in association with SCoV particles.


Personal protective equipment.


title: Nucleotide sequence of the gene encoding the surface projection glycoprotein of coronavirus MHV-JHM.

text: Sequences encoding the surface projection glycoprotein of the coronavirus, murine hepatitis virus (MHV), strain JHM, have been cloned into pAT153 using cDNA produced by priming with specific oligonucleotides on infected cell RNA. The regions of three clones pJMS1010, pJS112 and pJS92, which together encompass the surface protein gene have been sequenced by the chain termination method. The sequence of the primary translation product, deduced from the DNA sequence, predicts a polypeptide of 1,235 amino acids with a molecular weight of 136,600. This polypeptide displays the features characteristic of a group 1 membrane protein; an amino-terminal signal sequence and carboxy-terminal membrane and cytoplasmic domains. There are 21 potential glycosylation sites in the polypeptide and a cysteine-rich region in the vicinity of the transmembrane domain. During maturation proteolytic processing of the polypeptide occurs and at positions 624 to 628 the sequence Arg-Arg-Ala-Arg-Arg is found, which is similar to a number of basic sequences involved in the cleavage of enveloped RNA virus glycoproteins. The fusogenic properties of the MHV surface protein do not appear to correlate with a strongly hydrophobic region at the putative amino terminus of the carboxy-terminal cleavage product.


Coronaviruses, a new group of animal RNA viruses.


title: Cross-species transmission in the speciation of the currently known murinae-associated hantaviruses.

text: To gain more insight into the phylogeny of Dabieshan virus (DBSV), carried by Niviventer confucianus and other Murinae-associated hantaviruses, genome sequences of novel variants of DBSV were recovered from Niviventer rats trapped in the mountainous areas of Wenzhou, China. Genetic analyses show that all known genetic variants of DBSV, including the ones identified in this study, are distinct from other Murinae-associated hantaviruses. DBSV variants show geographic clustering and high intraspecies diversity. The data suggest that DBSV is a distinct species in the genus Hantavirus. Interestingly, DBSV shows the highest sequence identity to Hantaan virus (HTNV), with a >7% difference in the sequences of the N, GPC, and L proteins, while N. confucianus is more closely related to Rattus norvegicus (the host of Seoul virus [SEOV]) than to Apodemus agrarius (the host of HTNV and Saaremaa virus [SAAV]). Further genetic analyses of all known Murinae-associated hantaviruses (both established and tentative species) show that many of them, including DBSV, may have originated from host switching. The estimation of evolutionary rates and divergence time supports the role of cross-species transmission in the evolution of Murinae-associated hantaviruses. The detection of positive selection suggests that genetic drift may contribute to the speciation of Murinae-associated hantaviruses and that adaptation has a role as well.


title: Seroprevalence of selected infectious agents in a free-ranging, low-density lion population in the Central Kalahari Game Reserves in Botswana.

text: Twenty-one free-ranging Central Kalahari lions (Panthera leo) exhibited a high prevalence rate of feline herpesvirus (100%) and feline immunodeficiency virus (71.4%). Canine distemper virus and feline calicivirus occurred with a low prevalence. All individuals tested negative for feline coronavirus, feline parvovirus, feline leukemia virus, Ehrlichia canis, and Anaplasma phagocytophilum.


title: Pulmonary pathological features in coronavirus associated severe acute respiratory syndrome (SARS).

text: BACKGROUND Severe acute respiratory syndrome (SARS) became a worldwide outbreak with a mortality of 9.2%. This new human emergent infectious disease is dominated by severe lower respiratory illness and is aetiologically linked to a new coronavirus (SARS-CoV). METHODS Pulmonary pathology and clinical correlates were investigated in seven patients who died of SARS in whom there was a strong epidemiological link. Investigations include a review of clinical features, morphological assessment, histochemical and immunohistochemical stainings, ultrastructural study, and virological investigations in postmortem tissue. RESULTS Positive viral culture for coronavirus was detected in most premortem nasopharyngeal aspirate specimens (five of six) and postmortem lung tissues (two of seven). Viral particles, consistent with coronavirus, could be detected in lung pneumocytes in most of the patients. These features suggested that pneumocytes are probably the primary target of infection. The pathological features were dominated by diffuse alveolar damage, with the presence of multinucleated pneumocytes. Fibrogranulation tissue proliferation in small airways and airspaces (bronchiolitis obliterans organising pneumonia-like lesions) in subpleural locations was also seen in some patients. CONCLUSIONS Viable SARS-CoV could be isolated from postmortem tissues. Postmortem examination allows tissue to be sampled for virological investigations and ultrastructural examination, and when coupled with the appropriate lung morphological changes, is valuable to confirm the diagnosis of SARS-CoV, particularly in clinically unapparent or suspicious but unconfirmed cases.


title: Impact of COVID-19 Pandemic on Oncology Practices During Nationwide Lockdown Period: A Single Centre Experience and the Way Forward.

text: Background The COVID-19 pandemic has posed a grave challenge to mankind as it doesn't appear to get controlled in the near future. Worldwide, health care centers are working more than their capacity with the scarcity of medical resources. Cancer patients are considered to be at higher risk of developing life-threatening complications from COVID-19 and at the same time treatment delays can lead to poorer oncological outcomes. Appropriate planning is therefore important to continue with cancer treatment services and simultaneously avoiding the risk of infection to the patients and healthcare staff and not allowing community transmission of viral infection. Methods and Results We modified our practice measures in cancer patients receiving systemic therapy. Tele-consultations and use of electronic means, providing the best supportive care at or near home, and involvement of local/ family physicians were widely practiced. We minimised in-patient admissions, however, day-care chemotherapies were continued to provide optimum oncology services. Conclusions Modified oncological practice measures need to be implemented as the pandemic seems to stay for a longer time.


[Transanal endoscopic microsurgery (TEM) and transanal minimally invasive surgery (TAMIS)].


title: Coronavirus IBV glycopolypeptides: size of their polypeptide moieties and nature of their oligosaccharides.

text: Analysis of differentially radiolabelled avian infectious bronchitis virus (IBV) indicated that the matrix (M) polypeptides of mol. wt. 23 X 10(3) (23K), 26K, 28K, 30K and 34K (M23 to M34) which have been shown to give the same peptide maps, differed in their degree of glycosylation; M23 was not glycosylated while glycosylation increased with increasing mol. wt. from M26 to M34. Both glucosamine and mannose were components of M26 to M34 but [3H]fucose appeared to be associated mainly with M34. Endo-beta-N-acetylglucosaminidase H removed oligosaccharides from M28 and M30 but not M26 and M34, to give a polypeptide of 23K. The surface projection glycopolypeptides S1 (90K) and S2 (84K) incorporated 3H-labelled glucosamine and mannose but not fucose and had oligosaccharides removed by endoglycosidase H. The mol. wt. of the resultant polypeptides varied among experiments; the lowest mol. wt. observed were 64K and 61K. These results indicate (i) that the polypeptide moieties of the S polypeptides are approximately 64K and 61K, and 23K for the M polypeptide, (ii) that the oligosaccharides of the S and M polypeptides are of the high-mannose type and are linked to the polypeptides by N-glycosidic linkages, and (iii) that the M glycoprotein of IBV differs from that of murine coronaviruses and bovine coronavirus L9 which have O-linked oligosaccharides.


title: Engineered antibodies take center stage.

text: The start of the post-genomic era provides a useful juncture for reflection on the state of antibody engineering, which will be a critical technology for relating function and pathology to genomic sequence in biology and medicine. The phenomenal progress in deciphering the human genome has given significant impetus to the application of engineered antibodies in proteomics. Thus, advances in phage display antibody libraries can now help to define novel gene function and the measurement of abnormal protein expression in pathological states. Furthermore, intrabody and antibody engineering provide vehicles for the development of molecular medicines of the future. In addition to these new directions, antibody engineering has begun to show concrete success in its long-term efforts to develop targeted immunotherapies for cancer and other diseases. The cornerstones of clinical development are the detailed academic clinical trials that continue to push the boundaries of engineered antibodies into the real world. The field displays a healthy impatience for practical results, as research accelerates with concerted efforts to transfer preclinical insights into clinical trials. Growing private and governmental expenditures will lead to the rapid expansion of life-saving immunotherapeutic agents. The present review developed from our effort to report on the 11th Annual International Conference on Antibody Engineering (3-6 December 2000). This annual meeting is a forum for discussions on the latest advances in antibody engineering groups from around the world, and now includes the broader agenda of engineering in molecular immunology. In bringing scientists together to exchange ideas at this open forum, new collaborations and the threads of new discoveries are woven. For example, Professors Gerhard Wagner (Harvard Medical School), Dennis Burton (Scripps Research Institute), and Peter Hudson (CSIRO, Melbourne, Australia) gave exciting insights on structural immunobiology that had implications across many disciplines. The growth in antibody engineering was highlighted by the attendance of some 600 participants at the meeting, doubling that of the 1999 meeting. Dramatic clinical acceptance of monoclonal antibodies during the past two years has fostered this growth, with sales in 2000 of 1.8 billion dollars and projections for 2001 of 3 billion dollars. However, economic measures cannot begin to convey the medical revolution that is being effected by these first humanized and chimerized monoclonal antibodies. At this juncture, the 10 monoclonal antibody therapeutics in clinical use are of murine origin, of which 3 are entirely murine (OKT3, Mylotarg, 90Y-labeled Bexxar), 4 have been chimerized (human constant domains replacing murine) (ReoPro, Rituxan and its 131I-labeled analogue (Zevalin), Simulect, Remicade) and 3 were chimerized and humanized (human residues being substituted for at least some mouse-specific framework residues in VH and VL) (Zenapax, Herceptin, Synagis). Fully humanized anti-CD52 (CAMPATH-1H) has also been approved by the FDA for the treatment of B-cell chronic lymphocytic leukemia and should become available in late 2001. Humanization was initially developed by Dr. Greg Winter at the MRC Laboratory of Molecular Biology (Cambridge, UK), who presented the meeting's keynote address, "Antibodies as a Paradigm for Molecular Evolution". His pioneering work in antibody phage display libraries has been reformulated into a daring approach to develop truly novel proteins with genetically paired structural elements. He described studies in combinatorial protein engineering with enormous implications for both industrial and therapeutic applications of macromolecules.


title: Longitudinal profile of immunoglobulin G (IgG), IgM, and IgA antibodies against the severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein in patients with pneumonia due to the SARS coronavirus.

text: By using a recombinant severe acute respiratory syndrome coronavirus (SARS-CoV) nucleocapsid protein-based enzyme-linked immunosorbent assay (ELISA) and serum specimens serially collected (from day 0 to day 240 after symptom onset) from patients with pneumonia due to SARS-CoV, we analyzed the longitudinal profiles of immunoglobulin G (IgG), IgM, and IgA antibodies against the SARS-CoV nucleocapsid protein in patients with pneumonia due to SARS-CoV. For IgG, the median optical density at 450 nm (OD450) turned positive at day 17 and a biphasic response was observed. At day 240, all patients were still positive for anti-nucleocapsid protein IgG antibody. For IgM, the median OD450 turned positive at day 20.5, peaked at about day 80, and fell to below the baseline level at about day 180. At day 240, 36% of the patients were still positive for anti-nucleocapsid protein IgM antibody. For IgA, the median OD450 turned positive at day 17, peaked at about day 50, and fell to below the baseline level at about day 180. At day 240, 36% of the patients were still positive for anti-nucleocapsid protein IgA antibody. The time of seroconversion detected by the recombinant SARS-CoV nucleocapsid protein-based ELISA and that detected by indirect immunofluorescence assay were similar. The median times of seroconversion for IgG, IgM, and IgA detected by the indirect immunofluorescence assay were 17 days (17 days by ELISA), 16.5 days (20.5 days by ELISA), and 17.5 days (17 days by ELISA), respectively, after disease onset. One, four, and one of the six patients who died did not produce any IgG, IgM, and IgA antibodies against the nucleocapsid protein of SARS-CoV, respectively, although these antibodies were detected in all six patients by the indirect immunofluorescence assay. Further studies should be performed to see whether SARS-CoV nucleocapsid protein antibody positivity has any prognostic significance.


title: Valuation of the risk of SARS in Taiwan.

text: Two surveys conducted in Taiwan during the spring 2003 severe acute respiratory syndrome (SARS) epidemic reveal a high degree of concern about the threat posed by SARS to Taiwan and its residents, although respondents believe they are knowledgeable about the risk of SARS and that it is susceptible to individual control. Willingness to pay (WTP) to reduce the risk of infection and death from SARS is elicited using contingent valuation methods. Estimated WTP is high, implying values per statistical life of US dollars 3 to 12 million. While consistent with estimates for high-income countries, these values are substantially larger than previous estimates for Taiwan and may be attributable to the high degree of concern about SARS at the time the data were collected.


The accidental medical tourist.


title: Recent advances in diagnostic technology: applications in autoimmune and infectious diseases.

text: Biomarkers are used ubiquitously as indicators of biological health. The development of genomic and proteomic multiplex technologies have enormously amplified biomarker discovery and application to diagnostic and therapeutic decisions in clinical practice. New technologies are now available that simultaneously identify a wide spectrum of biomarkers and save time and costs. Multiplexed assays can be coupled to other disease specific indicators (i.e., cytokines, single nucleotide polymorphisms) in order to get more powerful information. However, there is an urgent need for validation/standardization of the new assays before they are adopted into clinical diagnostics. It is worthy to note a new assay, T cell interferon gamma release (TIGRAs), which has recently been introduced in the diagnosis of latent tuberculosis infection. It seems to perform better than tuberculin skin test in patients with inflammatory rheumatic diseases. In this review, we focus on advantages and limits of novel approaches to the detection of autoantibody profiles in autoimmune diseases or pathogen signatures in microbiology.


title: The Novel Use of a Low Prime Modified Ultrafiltration Apparatus in a 13-kg Jehovah's Witness Patient: A Case Report.

text: Modified ultrafiltration (MUF) is used in neonates and infants to reduce volume overload and increase oxygen-carrying capacity post cardiopulmonary bypass (CPB). In addition, it decreases edema, attenuates complementation activation and immunogenic response to CPB. Hemodilution in the pediatric patient has always been a challenge, countered in part by miniaturization of CPB circuits. We describe a case in which we maintained an acceptable hematocrit level greater than 24%, considered the nadir below which the adverse effects of hemodilution can become evident. We performed this by the novel use of an intravenous warming device (enFlow, Vyaire Medical, Mettawa, IL) to reduce the prime volume of our MUF circuit by more than 50%. We present the case and discuss the advantages and disadvantages of using a low-prime MUF circuit. We were able to conduct "bloodless" CPB, with the use of acute normovolemic hemodilution, miniaturization of the CPB and MUF circuits.


The practice of travel medicine: guidelines by the Infectious Diseases Society of America.


title: Influenza A (H1N1) was not associated with obesity in pregnant women living in Toluca, México.

text: OBJECTIVE The aim was to verify whether being overweight could have played a critical role in cases of mortality caused by influenza A (H1N1) in pregnant women. This virus' prevalence was also analyzed among people suffering from acute respiratory disease being attended at the state of Mexico's Autonomous University's medical research centre. METHODS The clinical files of women having influenza A (H1N1) attending the Monica Pretelini maternal-perinatal hospital's (HMPMP) intensive care unit in Toluca, Mexico, were reviewed. According to international recommendations, clinical detection of possible new cases of this disease was kept an open as a second step. RESULTS Five women suffering influenza A (H1N1) was attended at HMPMP's intensive care unit during 2009; only one survived. No differences in body mass index were found when comparing the anthropometric characteristics to another group of women affected by acute respiratory diseases; in fact, this parameter was below the limits for being overweight in both cases. No new case of influenza A (H1N1) was found after the first eight months of 2010. DISCUSSION It could not be verified whether being overweight was a factor of higher mortality due to influenza A (H1N1) amongst pregnant women in the state of Mexico. The key to better survival for pregnant women hospitalized with influenza A (H1N1) seemed to be early treatment with oseltamivir. The cases decreased dramatically after the severe wave of the new pandemic due to unknown reasons.


title: Zoonoses in solid-organ and hematopoietic stem cell transplant recipients.

text: Numerous reports exist of the transmission of zoonoses to humans during and after solid-organ and hematopoietic stem cell transplantation. Donor-derived infections of numerous etiologies, including West Nile virus infection, Chagas disease, toxoplasmosis, rabies, lymphocytic choriomeningitis virus infection, and infection due to Brucella species have been reported. Most zoonoses occur as a primary infection after transplantation, and immunocompromised patients are more likely to experience significant morbidity and mortality from these infections. Risks of zoonotic infection in the posttransplantation period could be reduced by patient education. Increased recognition of the risks of zoonoses, as well as the advent of molecular biology-based testing, will potentially augment diagnostic aptitude. Documented zoonotic infection as it affects transplantation will be the primary focus of this review.


title: Interaction of mouse hepatitis virus (MHV) spike glycoprotein with receptor glycoprotein MHVR is required for infection with an MHV strain that expresses the hemagglutinin-esterase glycoprotein.

text: In addition to the spike (S) glycoprotein that binds to carcinoembryonic antigen-related receptors on the host cell membrane, some strains of mouse coronavirus (mouse hepatitis virus [MHV]) express a hemagglutinin esterase (HE) glycoprotein with hemagglutinating and acetylesterase activity. Virions of strains that do not express HE, such as MHV-A59, can infect mouse fibroblasts in vitro, showing that the HE glycoprotein is not required for infection of these cells. The present work was done to study whether interaction of the HE glycoprotein with carbohydrate moieties could lead to virus entry and infection in the absence of interaction of the S glycoprotein with its receptor glycoprotein, MHVR. The DVIM strain of MHV expresses large amounts of HE glycoprotein, as shown by hemadsorption, acetylesterase activity, and immunoreactivity with antibodies directed against the HE glycoprotein of bovine coronavirus. A monoclonal anti-MHVR antibody, MAb-CC1, blocks binding of virus S glycoprotein to MHVR and blocks infection of MHV strains that do not express HE. MAb-CC1 also prevented MHV-DVIM infection of mouse DBT cells and primary mouse glial cell cultures. Although MDCK-I cells express O-acetylated sialic acid residues on their plasma membranes, these canine cells were resistant to infection with MHV-A59 and MHV-DVIM. Transfection of MDCK-I cells with MHVR cDNA made them susceptible to infection with MHV-A59 and MHV-DVIM. Thus, the HE glycoprotein of an MHV strain did not lead to infection of cultured murine neural cells or of nonmurine cells that express the carbohydrate ligand of the HE glycoprotein. Therefore, interaction of the spike glycoprotein of MHV with its carcinoembryonic antigen-related receptor glycoprotein is required for infectivity of MHV strains whether or not they express the HE glycoprotein.


European Resuscitation Council guidelines for resuscitation 2005. Section 4. Adult advanced life support.


title: Epidemiology of acute lung injury and acute respiratory distress syndrome.

text: Acute respiratory distress syndrome (ARDS) is a heterogeneous disorder that may be triggered by myriad etiologies (both pulmonary and extrapulmonary). Mortality rates for ARDS range from 30 to 75%, and most deaths are a consequence of multiorgan failure (MOF). Since the sentinel description of ARDS in 1967, criteria for the diagnosis of this entity were refined. In 1992, diagnostic criteria for ARDS were published in an American-European Consensus Conference and the term ACUTE LUNG INJURY (ALI) was adopted to encompass patients with a spectrum of less severe forms of the same pathological entity. This review discusses limitations of various criteria utilized to diagnosis ARDS and ALI, and why some criteria may be problematic when designing clinical trials. Also discussed are the myriad causes of ARDS, incidence, epidemiology, mortality, and factors that influence outcome.


title: Y box-binding protein-1 binds to the dengue virus 3'-untranslated region and mediates antiviral effects.

text: Dengue virus, a member of the family Flaviviridae, poses a serious public health threat worldwide. Dengue virus is a positive-sense RNA virus that harbors a genome of approximately 10.7 kb. Replication of dengue virus is mediated coordinately by cis-acting genomic sequences, viral proteins, and host cell factors. We have isolated and identified several host cell factors from baby hamster kidney cell extracts that bind with high specificity and high affinity to sequences within the untranslated regions of the dengue virus genome. Among the factors identified, Y box-binding protein-1 (YB-1) and the heterogeneous nuclear ribonucleoproteins (hnRNPs), hnRNP A1, hnRNP A2/B1, and hnRNP Q, bind to the dengue virus 3'-untranslated region. Further analysis indicated that YB-1 binds to the dengue virus 3' stem loop, a conserved structural feature located at the 3' terminus of the 3'-untranslated region of many flaviviruses. Analysis of the impact of YB-1 on replication of dengue virus in YB-1+/+ and YB-1-/- mouse embryo fibroblasts indicated that host YB-1 mediates an antiviral effect. Further studies demonstrated that this antiviral impact is due, at least in part, to a repressive role of YB-1 on dengue virus translation via a mechanism that requires viral genomic sequences. These results suggest a novel role for YB-1 as an antiviral host cell factor.


title: The 2006 American Association of Feline Practitioners Feline Vaccine Advisory Panel report.

text: Vaccination is a medical procedure, and the decision to vaccinate should be based on a risk-based assessment for each cat and each vaccine.


title: In-vitro Comparison of Antimicrobial Efficacy of Various Wound Dressing Materials.

text: UNLABELLED Many new antimicrobial dressings have been used for the treatment of or protection against wound infection since the early 1980s. There are many different wound dressings such as silver, povidone iodine, and chlorhexidine impregnated materials on the market today. Various manufacturers assert that their dressings are the most effective and therefore should be preferentially employed. However, it is difficult to find a study that clearly identifies the most effective antimicrobial wound dressing. METHODS Eight different commercially available wound dressing materials were compared in terms of their antimicrobial effectiveness on 18 different microorganisms via disk diffusion test (Kirby-Bauer Method) on Mueller-Hinton (MH) agar. Among the 16 bacterial and 2 yeast species grown on MH agar plates, Contreet was the most effective antimicrobial dressing tested (P <0.001). RESULTS A statistical difference was not found concerning efficacy against gram-positive and -negative bacteria among any of the materials except for Aquacel Ag and Inadine, the activity of which were found to be higher on gram-positive bacteria compared to gram-negative bacteria (P = 0.029, P = 0.030). In-vitro data suggest that Contreet is the most effective for topical treatment of contaminated wounds. CONCLUSION Further methods of assessment, including the use of infected animal models and clinical studies, will be necessary to better understand the antimicrobial efficacy of these dressings .


title: DC-SIGN and immunoregulation.

text: Dendritic cells (DCs) are known to be the most powerful professional antigen-presenting cells so far. It could activate primary immune response, and also downregulate immune response. DCs have a unique character of immunoregulation. DC-SIGN, a molecule designated as CD209, is one member of the C-type lectin superfamily. It is not only a pattern recognition receptor but implicated in immunoregulation of DCs. DC-SIGN has become hotspot of recent studies because of its important role in mediating DC adhesion, migration, inflammation, activating primary T cell, triggering immune response and participating in immune escape of pathogens and tumors. These studies on DC-SIGN involved in primary and secondary immune response and relevant mechanism will certainly provide us with a new method in treating and preventing certain diseases.


title: Reliability, validity and psychometric properties of the Greek translation of the Zung Depression Rating Scale.

text: INTRODUCTION The current study aimed to assess the reliability, validity and psychometric properties of the Greek translation of the Zung Depression Rating Scale (ZDRS). METHODS The study sample included 40 depressed patients 29.65 +/- 9.38 years old and 120 normal comparison subjects 27.23 +/- 10.62 years old. In 20 of them (12 patients and 8 comparison subjects) the instrument was re-applied 1-2 days later. Translation and Back Translation was made. Clinical Diagnosis was reached by consensus of two examiners with the use of the SCAN v.2.0 and the IPDE. Statistical Analysis included ANOVA, the Pearson Product Moment Correlation Coefficient, Principal Components Analysis and Discriminant Function Analysis and the calculation of Cronbach's alpha (alpha) RESULTS Both Sensitivity and specificity exceed 90.00 at 44/45, Chronbach's alpha for the total scale was equal to 0.09, suggesting that the scale covers a broad spectrum of symptoms. Factor analysis revealed five factors (anxiety-depression, thought content, gastrenterological symptoms, irritability and social-interpersonal functioning). The test-retest reliability was satisfactory (Pearson's R between 0.92). CONCLUSION The ZDRS-Greek translation is both reliable and valid and is suitable for clinical and research use with satisfactory properties. Its properties are similar to those reported in the international literature, although the literature is limited. However one should always have in mind the limitations inherent in the use of self-report scales.


title: [Isolation of cell hybrids secreting specific antibodies against bovine enteric coronavirus (author's transl)].

text: Twelve hybrid cell lines secreting monoclonal immunoglobulins (IgG) specific of bovine enteric Coronavirus (BEC strain G110) have been obtained following fusion of non secreting variant of the myeloma P3X63Ag8, SP2O, which splenic lymphocytes of Mice immunized against this virus. Antibodies thus obtained have been studied by indirect immunofluorescence and seroneutralization with cells infected by two french strains G110 and F15 and an american strain of Coronavirus: NCDCV. Immunochemical studies have confirmed this specificity.


title: Effect of single treatment with the antihypertensive drug eplerenone on hormone levels and anxiety-like behaviour in rats.

text: OBJECTIVE The aim of the study was to investigate the effects of single peripheral administration of mineralocorticoid receptor (MR) antagonist eplerenone on: 1. the hypothalamic-pituitary-adrenocortical (HPA) axis; 2. the renin-angiotensin-aldosterone (RAA) system, 3. anxiety-like behaviour. METHODS Male Wistar rats were injected subcutaneously with eplerenone (100 mg/kg body weight) or vehicle. Two hours following the injections, a half of the animals from each treatment group were decapitated to obtain blood for measurement of hormone levels. The other half of the animals was subjected to behavioural testing in the elevated plus-maze test. To provide comprehensive behavioural profile of eplerenone, standard spatiotemporal and ethologically derived measures of anxiety were assessed. RESULTS Single treatment with eplerenone resulted in a significant increase in plasma aldosterone levels. Plasma concentrations of ACTH and corticosterone were not modified by eplerenone injection. Administration of eplerenone failed to alter classical spatiotemporal measures of anxiety (number of entries and time spent in the open arms). However, ethological parameters related to exploration (head dipping) and risk assessment behaviour (stretched attend postures) were significantly affected by eplerenone injection. CONCLUSIONS Single injection of eplerenone is followed by a reduction of ethological indices of anxiety-like behaviour and by an elevation of plasma aldosterone levels. Acute administration of eplerenone appears to affect the RAA system but not hormones of the HPA axis.


title: [Clinical features of patients with influenza-like illness who went to a third level center in the winter of 2013-2014].

text: BACKGROUND Between October 2013 and April 2014, the Centers for Disease Control and Prevention reported a high incidence of AH1N1 influenza cases. Not all suspicious cases of influenza are confirmed; however, all patients need treatment and generate additional costs to the institutions. Our objective was to describe the characteristics of the patients treated for influenza suspicion at Hospital de Especialidades in Centro Médico Nacional Siglo XXI during a new epidemic. METHODS Cross-sectional study of all cases admitted at the Emergency Service (of the aforementioned hospital) for influenza-like illness, defined according to current guidelines. Based on these guidelines, we analyzed the risk factors that may have increased the severity of the infection. RESULTS We registered a total of 109 patients with a mean age of 44 years, 78 % were under 60 years of age, 62 % were women, 75 % had at least one comorbidity, such as obesity (26 %) or hypertension (27 %). Only 65 patients had results from a confirmatory test, 33.8 % had positive diagnosis, and 21 % of them eventually died. CONCLUSIONS The frequency of confirmed cases for influenza infection is low. The risk factors associated with complications and increased mortality are hypertension, leukocytosis and clinical presentation of severe acute respiratory syndrome.


title: Structural characterization of the SARS-coronavirus spike S fusion protein core.

text: The spike (S) glycoprotein of coronaviruses mediates viral entry into host cells. It is a type 1 viral fusion protein that characteristically contains two heptad repeat regions, denoted HR-N and HR-C, that form coiled-coil structures within the ectodomain of the protein. Previous studies have shown that the two heptad repeat regions can undergo a conformational change from their native state to a 6-helix bundle (trimer of dimers), which mediates fusion of viral and host cell membranes. Here we describe the biophysical analysis of the two predicted heptad repeat regions within the severe acute respiratory syndrome coronavirus S protein. Our results show that in isolation the HR-N region forms a stable alpha-helical coiled coil that associates in a tetrameric state. The HR-C region in isolation formed a weakly stable trimeric coiled coil. When mixed together, the two peptide regions (HR-N and HR-C) associated to form a very stable alpha-helical 6-stranded structure (trimer of heterodimers). Systematic peptide mapping showed that the site of interaction between the HR-N and HR-C regions is between residues 916-950 of HR-N and residues 1151-1185 of HR-C. Additionally, interchain disulfide bridge experiments showed that the relative orientation of the HR-N and HR-C helices in the complex was antiparallel. Overall, the structure of the hetero-stranded complex is consistent with the structures observed for other type 1 viral fusion proteins in their fusion-competent state.


title: Molecular approach to find target(s) for oligoclonal bands in multiple sclerosis.

text: OBJECTIVES Oligoclonal bands are a characteristic finding in the CSF of patients with multiple sclerosis, yet their target antigen(s) remain unknown. The objective was to determine whether a filamentous phage peptide library could be employed to allow the oligoclonal bands to select their own target epitopes. METHODS CSF IgG antibody from 14 patients with multiple sclerosis and 14 controls was used to select individual phage clones from a bacteriophage library containing approximately 4 x 10(7) different hexamers expressed on its surface pIII protein. The amino acid sequence selected was deduced by sequencing the DNA of the genetically engineered insert. RESULTS In general, after three rounds of selection, CSF from both patients with multiple sclerosis and controls selected one to two consistent peptide motifs. Five out of 14 patients with multiple sclerosis, and one control, selected the amino acid sequence motif, RRPFF. Given 20 possible amino acids per position, the likelihood of five patients selecting the same linear five amino acid sequence is at most 1.6 x 10(-3), corrected for the number of clones sequenced. A GenBank computer search showed that this sequence is found in the Epstein-Barr Virus nuclear antigen (EBNA-1), and a heat shock protein alphaB crystallin. Human serum antibodies to a synthetic peptide containing RRPFF were virtually exclusively found in patients with prior infection by Epstein-Barr virus. Other studies have suggested a relation between Epstein-Barr virus infection and multiple sclerosis, including nearly 100% Epstein-Barr virus seropositivity among patients with multiple sclerosis and increased concentrations of antibody to EBNA in CSF of patients with multiple sclerosis. By antigen specific immunoblotting, antibodies to the RRPFF motif in the CSF were shown to correspond to a subset of oligoclonal bands in the CSF from the same patient. CONCLUSION This study shows that phage epitope display libraries may be used to select amino acid motifs which are potentially relevant to the pathogenesis of multiple sclerosis.


title: [The virtual ward - the students' clinical education during the current pandemic].

text: On March 17, 2020, the Swedish Government recommended all higher education institutions to move to online and distance learning during the COVID-19 pandemic. The integrated course in Infection, Microbiology, and Immunity at the Programme in Medicine at University of Gothenburg had to be completely transformed. Creative solutions have now replaced the clinical training that normally takes place during the students' clinical education at the hospital. We developed a digital concept entitled "the virtual ward", in which we interact with the students in real time. Here, the students are able to follow their patients on a daily basis during teacher-guided sessions.


title: Coronavirus JHM: coding assignments of subgenomic mRNAs.

text: Protein synthesis in the murine hepatitis virus JHM-infected cells was temporarily inhibited by hypertonic shock. When the cells were returned to isotonic medium the synthesis of six virus-specific polypeptides, 150K, 65K, 60K, 30K, 23K and 14K was reinitiated simultaneously. Polyadenylated RNA isolated from the cytoplasm or polysomes of infected cells was translated in vitro and the products included polypeptides with molecular weights (mol. wt.) of 120,000, 60,000, 30,000, 23,000 and 14,000. Immunoprecipitation and fingerprinting of [35S]methionine-containing tryptic peptides showed that the 60,000 and 23,000 mol. wt. products were identical to the 60K and 23K polypeptides found in infected cells; the 120,000 mol. wt. product showed identity with the 150K intracellular polypeptide and a virus-specific 120K polypeptide synthesized in tunicamycin-treated cells. Two-dimensional polyacrylamide gel electrophoresis strongly suggested that the 30,000 and 14,000 mol. wt. products are equivalent to virus-specific 30K and 14K intracellular polypeptides. [3H]Uridine-labelled polyadenylated virus RNA was isolated from infected cells and sedimented in sucrose gradients containing formamide. The distribution in the gradient of each of the previously identified virus RNAs was determined by gel electrophoresis and gradient fractions enriched for each RNA were translated in vitro. The 120,000, 60,000, 30,000, 23,000 and 14,000 mol. wt. polypeptides were found to be encoded by mRNAs 3, 7, 2, 6, and 4 or 5 respectively. These results demonstrate that the virus-specific polypeptides in JHM-infected cells are encoded in separate subgenomic mRNAs and are translated independently. The assignment of coding functions and the known sequence relationships of JHM RNAs permitted a gene order to be deduced.


title: Covid 19 pandemic and gynaecological laparoscopic surgery: knowns and unknowns.

text: The worldwide impact of COVID 19 continues to be felt as hospitals in all countries reduce elective and non-urgent cases to allow staffing and resources to be deployed elsewhere. Urgent gynaecological and cancer procedures are continuing, and it is imperative all theatre staff are protected and risks of SARS-CoV-2 viral transmission reduced when operating on asymptomatic, suspected or confirmed COVID 19 patients. In particular, there are concerns relating to the transmission of COVID 19 during gynaecological laparoscopic surgery, arising from the potential generation of SARS-CoV-2 contaminated aerosols from CO2 leakage and the creation of smoke from the use of energy devices. The aim of this paper is to review all the up to date evidence, including experiences from China and Italy, to guide the safe management of such patients when undergoing gynaecological procedures.


title: Impact of a spreading epidemic on medical students.

text: The emergence of severe acute respiratory syndrome (SARS) had caused fear and anxiety of unprecedented proportion. To examine the impact of SARS on the medical students in a private medical university, a self-reporting questionnaire study was carried out to assess the factual knowledge, anxiety level and perception of the crisis, among the students. The two-week study (between 12 and 23 May, 2003) was carried out three weeks after the first reported SARS-related death in Malaysia. Ninety-one Phase I (junior) and 113 Phase II (senior) students completed the questionnaires. A large majority of students of Phase I and II were correct in their factual knowledge and were sensible in their perception of the future and the handling of the crisis by government(s). However, phase 1 students expressed significantly greater degree of anxiety compared to Phase II in relation to attendance and personal protection in hospital, and in meeting people coughing in public places. The lesser degree of anxiety expressed by phase II senior students may be due in part, to a more realistic assessment of SARS risk brought about by maturity, time spent in hospital and interaction with clinical lecturers and medical staff.


title: A tale of two cities: community psychobehavioral surveillance and related impact on outbreak control in Hong Kong and Singapore during the severe acute respiratory syndrome epidemic.

text: OBJECTIVES To compare the public's knowledge and perception of SARS and the extent to which various precautionary measures were adopted in Hong Kong and Singapore. DESIGN Cross-sectional telephone survey of 705 Hong Kong and 1,201 Singapore adults selected by random-digit dialing. RESULTS Hong Kong respondents had significantly higher anxiety than Singapore respondents (State Trait Anxiety Inventory [STAI] score, 2.06 vs 1.77; P < .001). The former group also reported more frequent headaches, difficulty breathing, dizziness, rhinorrhea, and sore throat. More than 90% in both cities were willing to be quarantined if they had close contact with a SARS case, and 70% or more would be compliant for social contacts. Most respondents (86.7% in Hong Kong vs 71.4% in Singapore; P < .001) knew that SARS could be transmitted via respiratory droplets, although fewer (75.8% in Hong Kong vs 62.1% in Singapore; P < .001) knew that fomites were also a possible transmission source. Twenty-three percent of Hong Kong and 11.9% of Singapore respondents believed that they were "very likely" or "somewhat likely" to contract SARS during the current outbreak (P < .001). There were large differences between Hong Kong and Singapore in the adoption of personal precautionary measures. Respondents with higher levels of anxiety, better knowledge about SARS, and greater risk perceptions were more likely to take comprehensive precautionary measures against the infection, as were older, female, and more educated individuals. CONCLUSION Comparative psychobehavioral surveillance and analysis could yield important insights into generic versus population-specific issues that could be used to inform, design, and evaluate public health infection control policy measures.


title: Development of one-step, real-time, quantitative reverse transcriptase PCR assays for absolute quantitation of human coronaviruses OC43 and 229E.

text: The clinical significance of human coronaviruses in more severe respiratory illnesses has recently been shown to be higher than was previously assumed. Rapid and reliable diagnosis of human coronavirus infections therefore becomes indispensable in a routine clinical setting. In this study, we present a very sensitive and specific TaqMan-based, real-time quantitative reverse transcriptase PCR (qRT-PCR) for the rapid detection and quantitation of human coronaviruses (HCoVs) OC43 and 229E. Absolute viral load measurement in clinical samples was achieved through the construction of in-house HCoV OC43 and 229E cRNA standards for the generation of a standard curve. The HCoV OC43 assay allows quantitation over a range from 20 to 2 x 10(8) RNA copies per reaction mixture (5 microl RNA extract). When this is extrapolated to clinical samples, this corresponds to a detection range of 10(3) to 10(10) viral genome equivalents per ml. By using the HCoV 229E qRT-PCR assay, viral RNA copies ranging from 200 to 2 x 10(9) per reaction mixture can be detected, which corresponds to 10(4) to 10(11) viral genome equivalents per ml sample. A total of 100 respiratory samples screened for the presence of HCoVs OC43 and 229E by using conventional RT-PCR were assessed in parallel by the qRT-PCR assays. By use of the real-time qRT-PCR techniques, the detection rate of HCoVs OC43 and 229E increased from 2.0% to 3.1% and from 0.3% to 2.5%, respectively. The real-time qRT-PCR assays described here allow the rapid, specific, and sensitive laboratory detection and quantitation of human coronaviruses OC43 and 229E.


title: Screening of respiratory pathogens by Respiratory Multi Well System (MWS) r-gene™ assay in hospitalized patients.

text: Novel respiratory viruses have been identified as possible agents of upper and lower respiratory tract infections. Multiplex real-time PCRs have been developed to identify clinically relevant respiratory pathogens. In this study, 178 respiratory samples already screened for influenza virus types A and B by Flu A/B ASR real-time PCR kit were retrospectively analyzed with the Respiratory Multi Well System (MWS) r-gene™ real-time PCR kit which detects a wide spectrum of respiratory pathogens. The goal was to demonstrate the importance of a wide spectrum screening compared to a single diagnostic request. The Flu A/B ASR kit detected influenza B virus in 1.7% of the samples (3/178) and no influenza A virus. The MWS r-gene™ kit detected influenza virus in 6.7% (12/178) of samples (0.6% influenza A, and 6.2% influenza B), while the overall detection rate for respiratory pathogens was 54% (96/178). Co-infections were detected in 8/178 (4.5%) samples. Adenovirus was the infectious agent detected most frequently, followed by respiratory syncytial virus. The risk of being infected by respiratory syncytial virus is almost threefold higher in patients older than 65 years compared to the younger age group (OR:2.7, 95% CI: 1.2-6.2). Wide spectrum screening of respiratory pathogens by real-time PCR is an effective means of detecting clinically relevant viral pathogens.


title: Novel aspects of the renin-angiotensin-aldosterone-system.

text: The renin-angiotensin-aldosterone system (RAAS) play a pivotal role in the progression of renal disease. The RAAS has become much more complex in recent years with the identification of novel peptides that exhibit biological activity. There are novel pathways of angiotensin II (ANG II) generation independent of angiotensin converting enzyme (ACE). ANG II bind to at least two different receptors and prorenin/renin also exerts pathophysiological effects through binding to specific receptor. ANG II itself has emerged as a multifunctional cytokine exhibiting many non-hemodynamic properties such as acting as a growth factor and profibrogenic and proinflammatory cytokine. These profibrogenic and proinflammatory effects are mediated by other factors such as transforming growth-factor beta (TGF-beta) and chemoattractants that are induced in the kidney by ANG II. Increased aldosterone levels contribute to renal injury, independent of blood pressure or ANG II. Numerous experimental and clinical studies have shown that ACE-inhibitors as well as AT1-receptor antagonists can prevent glomerulosclerosis and tubulointerstitial fibrosis. This review will highlight some of these novel insights into the RAAS in regards to renal injury.


title: Echocardiographic assessment of coronary artery flow in normal canines and model dogs with myocardial infarction.

text: This study was conducted to evaluate the usefulness of coronary arterial profiles from normal dogs (11 animals) and canines (six dogs) with experimental myocardial infarction (MI) induced by ligation of the left coronary artery (LCA). Blood velocity of the LCA and right coronary artery (RCA) were evaluated following transthoracic pulsed-wave Doppler echocardiography. The LCA was observed as an infundibular shape, located adjacent to the sinus of Valsalva. The RCA appeared as a tubular structure located 12 o'clock relative to the aorta. In normal dogs, the LCA and RCA mean peak diastolic velocities were 20.84 ± 3.24 and 19.47 ± 2.67 cm/sec, respectively. The LCA and RCA mean diastolic deceleration times were 0.91 ± 0.14 sec and 1.13 ± 0.20 sec, respectively. In dogs with MI, the LCA had significantly (p < 0.01) lower peak velocities (14.82 ± 1.61 cm/sec) than the RCA (31.61 ± 2.34 cm/sec). The RCA had a significantly (p < 0.01) rapid diastolic deceleration time (0.71 ± 0.06 sec) than that found in the LCA (1.02 ± 0.22 sec) of MI dogs. In conclusion, these profiles may serve as a differential factor for evaluating cardiomyopathy in dogs.