title: Passive protection against porcine epidemic diarrhea (PED) virus in piglets by colostrum from immunized cows.

text: The effects of hyperimmune cow colostrum (HCC) on experimentally induced porcine epidemic diarrhea (PED) were investigated in piglets. In experiment 1, four 2-day-old piglets fed HCC containing an antibody titer of 1:512 and another four piglets fed unimmune cow colostrum (UCC) were orally inoculated with 10LD50 of PED virus. The piglets were given colostrum three times a day at 4 hr intervals. Half of the piglets fed HCC showed diarrhea and recovered, and all piglets survived. In contrast, all piglets fed UCC developed diarrhea and three of them died. In experiment 2, 2-day-old piglets fed HCC containing antibody titers of 1:512, 1:128 and 1:32, and UCC were inoculated with PED virus, and survival rates after challenge were 100, 75, 50 and 0 %, respectively. In experiment 3, 1-day-old piglets fed HCC with 1:512 antibody titer or UCC were inoculated and necropsied at 24, 48 and 72 hr after the inoculation for pathological examination. Piglets fed HCC remained healthy and PED virus antigen was not detected in the epithelial cells of the small intestine, and the length of the villi in small intestine was normal. On the other hand, in piglets fed UCC, villous atrophy and PED virus antigen were observed in epithelial cells of the jejunum and ileum from 24 hr. It was concluded that oral administration of HCC to piglets was effective in preventing PED virus infection and reduced their mortality.


title: Evaluation of laparoendoscopic single-site gynecologic surgery with a multitrocar access system.

text: OBJECTS Laparoendoscopic single-site surgery (LESS) is an ideal approach for gynecologic surgery and yields better cosmetic results. However, a standard umbilical approach with LESS is not appropriate for gynecologic surgery requiring intra-abdominal suturing and dissection requiring traction. Therefore, we have developed a new multitrocar access system for gynecologic LESS. The purpose of this study was to evaluate the efficacy of gynecologic LESS using this access system. METHODS This access system consists of one 12-mm trocar, two 5-mm trocars, and a 5-mm flexible laparoscope. Two 5-mm trocars with small port heads were inserted cross-wise on opposite sides of the sleeve of the centrally positioned 12-mm trocar to maintain triangulation. Thirty-eight patients with various gynecologic conditions underwent LESS with this access system. The results of these surgeries were retrospectively compared to those of conventional laparoscopic procedures. RESULTS Of the 38 LESS procedures performed with this access system, none was up-converted, converted to an open laparotomy, or required blood transfusion. The Salpingo-oophorectomy with LESS had several benefits, such as no extension of the skin incision of the trocar site and no leakage of the contents of the ovarian cyst into the peritoneal cavity, over that with conventional laparoscopy. A comparison of LESS (11 patients) and conventional laparoscopy (16 patients) for total hysterectomy showed no significant difference in total blood loss (234.0 mL vs. 221.6 mL) or the weight of the resected uterus (276.0 g vs. 285.0 g), although the mean total operative time was greater with LESS (199.0 min vs. 168.5 min). CONCLUSION Our multitrocar access system is safe and secure, and can be adapted for various gynecologic surgeries involving complicated procedures. LESS with this access system achieves results comparable to those of conventional laparoscopy with 4 ports, although the operative time is longer.


title: Nurses and the control of infectious disease. Understanding epidemiology and disease transmission is vital to nursing care.

text: Epidemiology examines the distribution and source of a disease in a population. Understanding epidemiology and disease transmission is vital to nursing care. Infectious disease transmission requires three components: an agent (virus, bacterium, parasite or other microbe), a vulnerable host and a conducive environment. Disease spread can occur through direct contact or via indirect methods (airborne droplets, vectors, fomites, water or food). Intervention can occur by attacking the agent (e.g., using microbicides), changing the environment (e.g., providing negative pressure rooms) or strengthening the host (e.g., vaccination). Three epidemiologically relevant microbes are the SARS (severe acute respiratory syndrome)-associated coronavirus, methicillin-resistant Staphylococcus aureus (MRSA) and Clostridium difficile (C. difficile). The first is an emerging pathogen, and the latter two are existing agents that have mutated such that they are resistant to their standard treatments. For SARS, control measures include screening for possible cases and appropriate triage, respiratory and barrier precautions within the healthcare facility, and voluntary isolation in the community for contacts or healthcare workers who exhibit symptoms. Control measures for MRSA include the screening of patient lesions, isolating or cohorting patients who are already infected, covering wounds with impermeable dressings, treating staff and patient carriers with antibiotics, and improved hygiene. Control measures for C. difficile Control measures include paying close attention to the hygiene of the clinical setting, disinfecting using bleach and the isolation of infected patients.


title: Chylous ascites post open cholecystectomy after severe pancreatitis.

text: CONTEXT Chylous ascites a rare complication post cholecystectomy. There are to our knowledge only 3 reported cases in the literature. We describe a case of chylous ascites post open cholecystectomy in a patient with recent severe pancreatitis. We propose a potential relationship between acute biliary pancreatitis and the development of chylous ascites. CASE REPORT We present a 37 year old man with a history of severe biliary pancreatitis 10 months prior. He was electively admitted for laparoscopic cholecystectomy. His surgery was converted to open cholecystectomy due to adhesions. A closed, non suction drain was placed intraoperatively draining serosanguinous fluid. Two days post operatively the draining fluid changed to a milky colour fluid with high triglyceride level. Chylous ascites was confirmed clinically with a 24 hour fast. He was treated with total parenteral nutrition. His chylous ascites resolved with conservative management and his drain was removed. CONCLUSION We describe the diagnosis and management of chylous ascites post cholecystectomy. We propose the hypothesis to the aetiology of this rare condition, based on ours and one previous case, that a history of recent severe acute pancreatitis poses a risk factor in the development of chylous ascites.


Creating healthy, just and eco-sensitive cities.


Summary of the coronavirus disease 2019 (COVID-19) update from the 2020 Conference on Retroviruses and Opportunistic Infections, 8-11 March 2020, Boston, USA.


title: [Disseminated pulmonary infection due to Cryptococcus neoformans in a non immunocompromised patient].

text: We report a case of a primary pulmonary infection due to Cryptococcus neoformans developed in a 40 Year old immunocompetent and HIV negative man. Radiologic findings consisted in diffuse, bilateral reticular and nodular opacities. Bronchoscopy was normal and bronchoalveolar lavage showed numerous macrophages without associated pathogens. A thoracovideoscopy with an open lung biopsy showed numerous cryptococci, free in the alveoli or located within the macrophages. They were associated with an inflammatory infiltrate in the interalveolar spaces, predominantly composed of mononuclear cells. Ultrastructural study showed yeasts with numerous intracytoplasmic organites, a nucleus, a wall and a thick capsule. Pulmonary cryptococcosis is rare in immunocompetent hosts and can be difficult to diagnose since clinicoradiologic features observed in this cryptococcal infection mimic other infectious or neoplastic diseases.


title: The peplomer protein E2 of coronavirus JHM as a determinant of neurovirulence: definition of critical epitopes by variant analysis.

text: We selected murine coronavirus JHM variants specifically changed in defined antigenic sites of the peplomer protein E2. Variants were isolated from the supernatants of monoclonal antibody hybridoma cell cultures which continued to secrete neutralizing antibodies after being infected with JHM. Comparative antigenic analysis and biological tests were performed in order to refine an operational epitope map and to characterize functional domains important for pathogenicity. The reaction patterns (neutralization, inhibition of cell fusion, immunofluorescence and binding in ELISA) between the variant viruses and the panel of monoclonal antibodies were very similar. Four groups of variants were characterized each of which revealed distinct changes affecting one defined antigenic site. These observations indicated that at least four independently mutable antigenic sites were associated with domains involved in cell fusion, neutralization and pathogenicity (E2-Aa, -Ab, -Ba and -Bb). JHM variants with alterations in the E2-Aa, -Ab or -Bb sites were similar to wild-type virus. These variants caused acute hepatitis and encephalomyelitis in mice. In contrast, JHM variants with changes in site E2-Ba had a strong propensity to induce chronic disease accompanied by demyelination persisting for several months.


title: Characteristics, treatment, outcomes and cause of death of invasively ventilated patients with COVID-19 ARDS in Milan, Italy.

text: Objective Describe characteristics, daily care and outcomes of patients with coronavirus disease 2019 (COVID-19) acute respiratory distress syndrome (ARDS). Design Case series of 73 patients. Setting Large tertiary hospital in Milan. Participants Mechanically ventilated patients with confirmed COVID-19 admitted to the intensive care unit (ICU) between 20 February and 2 April 2020. Main outcome measures Demographic and daily clinical data were collected to identify predictors of early mortality. Results Of the 73 patients included in the study, most were male (83.6%), the median age was 61 years (interquartile range [IQR], 54-69 years), and hypertension affected 52.9% of patients. Lymphocytopenia (median, 0.77 x 103 per mm3 ; IQR, 0.58-1.00 x 103 per mm3), hyperinflammation with C-reactive protein (median, 184.5 mg/dL; IQR, 108.2-269.1 mg/dL) and pro-coagulant status with D-dimer (median, 10.1 μg/m; IQR, 5.0-23.8 μg/m) were present. Median tidal volume was 6.7 mL/kg (IQR, 6.0-7.5 mL/kg), and median positive end-expiratory pressure was 12 cmH2O (IQR, 10-14 cmH2O). In the first 3 days, prone positioning (12-16 h) was used in 63.8% of patients and extracorporeal membrane oxygenation in five patients (6.8%). After a median follow-up of 19.0 days (IQR, 15.0-27.0 days), 17 patients (23.3%) had died, 23 (31.5%) had been discharged from the ICU, and 33 (45.2%) were receiving invasive mechanical ventilation in the ICU. Older age (odds ratio [OR], 1.12; 95% CI, 1.04-1.22; P = 0.004) and hypertension (OR, 6.15; 95% CI, 1.75-29.11; P = 0.009) were associated with mortality, while early improvement in arterial partial pressure of oxygen (PaO2) to fraction of inspired oxygen (FiO2) ratio was associated with being discharged alive from the ICU (P = 0.002 for interaction). Conclusions Despite multiple advanced critical care interventions, COVID-19 ARDS was associated with prolonged ventilation and high short term mortality. Older age and pre-admission hypertension were key mortality risk factors. Trial registration ClinicalTrials.gov identifier: NCT04318366.


title: [Present state of in vitro and in vivo studies on efficacy of medical nonspecific protective agents with respect to genotype IV coronavirus causing severe acute respiratory syndrome].

text: The results of the in vitro studies on the efficacy of medical nonspecific protective agents from various pharmacological groups showed that some drugs, such as velferon, alferon, betaferon, ribavirin and lopinavir were active against TOPC virus, that permitted to recommend them for estimation of their activity on laboratory animals. The data on the in vivo activity of pharmacological drugs with respect to TOPC virus are rather scanty and it is difficult to predetermine their efficacy. The danger of TOPC virus latent circulation among wild animals in China requires research of new efficient medical agents for protection of the people from the pathogen in the Russian Federation.


title: Respiratory viral infections in adults with and without chronic obstructive pulmonary disease.

text: A longitudinal cohort study of older adults with chronic obstructive pulmonary disease (COPD) who were stratified by FEV(1) at enrollment was done to define the etiology, frequency, severity, and medical-care impact of respiratory tract viral infections (RTVIs). Controls consisted of a group of subjects of comparable age with the patients. RTVIs were documented in 44% of observed acute respiratory illnesses in control subjects and in 27% of COPD subjects, who were followed for mean periods of 35 and 26 mo, respectively. In this heavily influenza-vaccinated cohort ( approximately 90% vaccinated each year), picornaviruses, parainfluenza viruses, and coronaviruses were most commonly identified. Mean time to return to clinical baseline was approximately 2 wk in each group. Control and COPD subjects with mild airways obstruction (baseline FEV(1) >/= 50% predicted) had few emergency-center visits or hospitalizations. Approximately half of COPD subjects with moderate/severe COPD (baseline FEV(1) < 50% predicted) had at least one emergency-center visit and/or hospitalization for acute respiratory illness. RTVIs were documented in 23% of hospitalizations and in 45% of patients admitted between December and March. RTVIs have a major impact on utilization of health care resources for COPD patients with moderate/severe airways obstruction.


title: Laparoendoscopic single-site adrenalectomy sans transumbilical approach: initial experience in Japan.

text: PURPOSE The use of laparoendoscopic single-site (LESS) surgery has been increasing. The LESS procedure has been done for various urological diseases and studies have shown that it results in less pain, shorter hospital stays and excellent cosmetic outcomes. MATERIALS AND METHODS We describe our initial experience with LESS adrenalectomy without the use of the transumbilical approach. The participants were 16 consecutive patients who underwent LESS adrenalectomy using a SILS port™ (Covidien, Mansfield, MA, USA) at Hiroshima University Hospital. Various parameters including the insufflation time, estimated blood loss, resumption of oral intake and complications were analyzed. RESULTS The adrenalectomy was completed successfully with no major intraoperative complications in 15 of the patients. One LESS adrenalectomy was converted to a conventional laparoscopic adrenalectomy by placement of two additional 12 mm trocars; this patient was therefore excluded from the study. The mean (range) patient age was 53.8 (35-69) years, body mass index (BMI) was 23.5 (20.7-27.2) kg/m2, tumor size was 19.8 (9-45) mm, insufflation time was 188.0 (95-340) min and estimated blood loss was 36.3 (10-80) mL. In all cases articulating instruments were used for satisfactory dissection and triangulation. One bowel injury (serosal) occurred and was repaired in the open laparotomy before SILS port™ insertion. All patients at the follow-up visit were satisfied and pleased with their scars. CONCLUSION We found LESS adrenalectomy to be apparently safe, effective and minimally invasive for adrenal diseases. In our opinion, LESS adrenalectomy without using the transumbilical approach is feasible and easy to introduce.


title: Evaluation of a computer-assisted, kinetics-based enzyme-linked immunosorbent assay for detection of coronavirus antibodies in cats.

text: A computer-assisted, kinetics-based enzyme-linked immunosorbent assay was adapted for the detection of coronavirus antibodies in feline serum. An alkaline antigen diluent (carbonate-bicarbonate buffer, pH 9.6) used in initial experiments produced diffuse, nonspecific color reactions in both viral and control antigen cuvettes which were correlated, paradoxically, with coronavirus antibody levels in test sera. These interfering reactions were minimized by use of lower-pH antigen diluents such as water and phosphate-buffered saline. Background kinetics-based enzyme-linked immunosorbent assay reactivity directed against a noncoronaviral component of antigen tissue culture fluids could then detected in numerous sera, particularly in samples with lower titers. Much of this reactivity was shown to be associated with bovine gamma globulins in cell culture fluid. It was not serum lot or species specific, since a variety of bovine serum lots as well as individual lots of serum from other mammalian and avian species reacted. Reactivity was markedly reduced when cells for antigen preparation were grown in gamma globulin-free bovine serum. Generation of corrected slope values from the kinetics-based enzyme-linked immunosorbent assay made it possible to correct for residual background reactivity in individual test sera and thus eliminate a potentially major source of false-positive reactions. Collectively, these studies indicated that the control of nonspecific reactivity in feline coronavirus serology is absolutely essential to obtain useful estimates of specific antibody responses.


title: Total laparoscopic Wertheim's radical hysterectomy versus Wertheim's radical abdominal hysterectomy in the management of stage I cervical cancer in Singapore: a pilot study.

text: INTRODUCTION For many decades, Wertheim's radical hysterectomy via laparotomy, also known as Wertheim's radical abdominal hysterectomy (RAH), has been the traditional surgical approach for operable stage IB cervical cancer. However, many established cancer centres worldwide have recently shown total laparoscopic Wertheim's radical hysterectomy (TLRH) to be a safe and feasible alternative to the conventional abdominal route for early cervical cancer management. This technique was introduced in Singapore in 2009. METHODS This was a prospective pilot study comparing TLRH with RAH in a single large tertiary institution in Singapore. Inclusion criteria included surgically fit patients with early cervical cancer and no radiological evidence of regional or distant metastases. RESULTS From November 2009 to February 2011, a total of 18 TLRHs and 30 RAHs were performed. The median blood loss in the TLRH group was significantly lower than that in the RAH group (300 mL vs. 500 mL; p = 0.04). However, there was no statistically significant difference found between the two techniques in terms of operative time, hospital stay, bladder recovery, total lymph node yield or adjuvant treatment. No intraoperative bladder, ureteric or bowel complications were observed in the two groups. Postoperative complications occurred in 2 (11.1%) TLRH patients and 4 (13.3%) RAH patients. With a median follow-up of 37.3 (range 10-68) weeks, the rate of recurrence was found to be 5.6% for the TLRH group and 10.0% for the RAH group. CONCLUSION The results of our study suggest that with appropriate patient selection and increased experience, TLRH can be a safe and effective procedure for the management of early cervical cancer in Singapore.


Current literature in mass spectrometry.


title: Effects of modification of the transcription initiation site context on citrus tristeza virus subgenomic RNA synthesis.

text: Citrus tristeza virus (CTV), a member of the Closteroviridae, has a positive-sense RNA genome of about 20 kb organized into 12 open reading frames (ORFs). The last 10 ORFs are expressed through 3'-coterminal subgenomic RNAs (sgRNAs) regulated in both amounts and timing. Additionally, relatively large amounts of complementary sgRNAs are produced. We have been unable to determine whether these sgRNAs are produced by internal promotion from the full-length template minus strand or by transcription from the minus-stranded sgRNAs. Understanding the regulation of 10 sgRNAs is a conceptual challenge. In analyzing commonalities of a replicase complex in producing so many sgRNAs, we examined initiating nucleotides of the sgRNAs. We mapped the 5' termini of intermediate- (CP and p13) and low- (p18) produced sgRNAs that, like the two highly abundant sgRNAs (p20 and p23) previously mapped, all initiate with an adenylate. We then examined modifications of the initiation site, which has been shown to be useful in defining mechanisms of sgRNA synthesis. Surprisingly, mutation of the initiating nucleotide of the CTV sgRNAs did not prevent sgRNA accumulation. Based on our results, the CTV replication complex appears to initiate sgRNA synthesis with purines, preferably with adenylates, and is able to initiate synthesis using a nucleotide a few positions 5' or 3' of the native initiation nucleotide. Furthermore, the context of the initiation site appears to be a regulatory mechanism for levels of sgRNA production. These data do not support either of the established mechanisms for synthesis of sgRNAs, suggesting that CTV sgRNA production utilizes a different mechanism.


title: [Venous thrombotic risk related to SARS-CoV-2 : prevalence, recommendations and perspectives].

text: Since the SARS-CoV-2 pandemic outbreak, growing evidence suggests that patients suffering from COVID-19 are at increased risk of thrombotic events. The sepsis-related activation of the coagulation combined with a high prevalence of common thrombotic risk factors could contribute to this prothrombotic state. Coagulation biomarkers could help in the identification of patients at risk of complications and mortality. The incidence of venous thromboembolic events appears to be increased, especially in severe COVID-19 patients. Based on that knowledge, several societies have provided recommendation on the prevention of venous thromboembolism. In this narrative review, we summarize available epidemiologic data on venous thromboembolism and recommendations on thromboprophylaxis in COVID-19.


title: [Analyses of subset, activated state and expression pattern of 24 repertoire TCR Vbeta of peripheral blood T lymphocytes in convalescence patients with severe acute respiratory syndrome (SARS)].

text: AIM To study the function of lymphocytes and the expression pattern of 24 repertoire TCR Vbeta in the convalescent patients with severe acute respiratory syndrome(SARS) after treatment by combination of traditional Chinese and Western medicine. METHODS T lymphocyte subsets, activated states of T and B lymphocytes, and expressions of CD3(+) and 24 TCR Vbeta subfamilies in 76 convalescent SARS patients were detected by flow cytometry. RESULTS The percentages of CD3(+) and CD4(+) T cells were lower than normal reference (P<0.05 and P<0.01, respectively). The percentage of CD8(+) T lymphocytes was higher than normal reference (P<0.01),while the percentage of CD8(+)/CD28(-) T cells (Ts) in 49 convalescent SARS patients increased, and that of CD8(+)/CD28(+) T cells (Tc) in 39 patients decreased. The percentage of activated T cells (Ta,CD3(+)/HLA-DR(+)) in 36 convalescent SARS patients and the percentage of activated B cells (Ba, CD3(-)/HLA-DR(+)) in 30 convalescent SARS patients were higher than normal references. Expression of TCR Vbeta14 was the commonest in 24 TCR Vbeta repertoire both in SARS patients and normal controls, followed by TCR Vbeta5.3 in SARS patients, but Vbeta20 in normal controls. Besides, the expressions of TCR Vbeta1, Vbeta5.2, Vbeta5.3, Vbeta7.2, Vbeta9, Vbeta11, Vbeta13.1, Vbeta13.2, Vbeta17, Vbeta18, Vbeta22 and Vbeta23 in SARS group were all significantly higher than those in normal group. And the expressions of TCR Vbeta4, Vbeta22 and Vbeta23 in SARS patients treated with glucocorticoid(>1 000 U) were higher than those in SARS patients without treatment. CONCLUSION Increased Ts cell may lead to increased CD8(+) T cells in convalescent SARS patients. The suppressor factors secreted by Ts cells may decrease CD3(+) and CD4(+) T cells. Expression of TCR Vbeta on T cells from convalescent SARS patients were different from normal controls. The increased Ta and Ba expressions may be associated with activation of T and B cells.


title: Determination of the 5' end of the lactate dehydrogenase-elevating virus genome by two independent approaches.

text: We have determined the 5' end of the lactate dehydrogenase-elevating virus (LDV) genome (strain LDV-P) using two independent approaches. In one approach, methylmercuric hydroxide-denatured genomic RNA was reverse-transcribed using as primer an oligonucleotide complementary to the 5' end of open reading frame (ORF) 1a. The first-strand cDNA was ligated with T4 RNA ligase to an oligonucleotide of which the 3' end was blocked. The ligated product was amplified by PCR, cloned and sequenced. In the second approach, untreated or decapped genomic RNA was ligated between the 3' and 5' ends, reverse-transcribed across the ligation junction and the product was amplified by PCR, cloned and sequenced. Both approaches yielded the same results, indicating that the 5' leader of LDV-P is 156 nucleotides long, inclusive of the 5' UAUAACC 3' sequence involved in the linkage of the 5' leader to the bodies of the seven subgenomic mRNAs of LDV. The 5' leader of LDV is about 50 nucleotides shorter than those of the related viruses, equine arteritis virus and Lelystad virus, but at least twice as long as the leaders of the coronaviruses. The finding that untreated LDV RNA was ligated 5' to 3' end as efficiently as RNA treated with decapping enzyme suggests that genomic LDV RNA may not possess a 5' cap but terminates with 5' phosphoryl-A.


title: Retrospective study of bovine neonatal mortality: cases reported from INTA Balcarce, Argentina.

text: A retrospective study was performed on 169 beef and dairy calves aged from 1 to 7 days old submitted to the Diagnostic Laboratories at INTA Balcarce, Argentina. Bacterial culture was performed for aerobic and microaerophilic organisms. Samples from spleen and lymph nodes, and peripheral blood mononuclear cells were also cultured for viral isolation on cell culture. Bovine rotavirus was detected by direct-ELISA. Multiple tissue samples were fixed in 10% formalin, routinely processed and Stained with hematoxylin and eosin for microscopic examination. Etiological diagnosis was made in 70 of the 169 calves. Infectious agents were identified in 49 cases, the most common being Escherichia coli. When the histopathological examination was performed in cases with undetermined diagnosis, it was noted that 44 specimens had histological lesions, which suggested the presence of an infectious agent. In order to characterize the causes of bovine neonatal mortality, the protocols and methodology should be improved in further works.


title: Tips and tricks to avoid bile duct injury in SILC: an experience of 500 cases.

text: INTRODUCTION Conventional laparoscopic cholecystectomy is the gold standard surgical treatment for symptomatic gallstones. Surgeons have attempted to minimize the number of incisions via single-incision laparoscopic cholecystectomy (SILC), which offers benefits including improved cosmesis, possibly less postoperative pain, and improved patient satisfaction. However, studies show that there is an increased risk of operative complications-in particular bile duct injuries. We report 500 consecutive cases of SILC performed without bile duct injury. METHODS A retrospective study of 500 continuous cases of SILC performed by the same surgeon at a single institution was conducted. Data on patient demographics, operative details, and postoperative outcomes were collected and evaluated. Detailed analysis of surgical techniques specifically to reduce bile duct injury was performed and described in this study. RESULTS In total, 500 patients underwent SILC during the study period. Eight patients needed additional ports to complete the surgery, while one was converted to an open surgery. No serious intraoperative complications, such as bile duct injury, were encountered. CONCLUSION Our experience shows that with due care and caution during SILC, with particular attention towards achieving the critical view of safety and a standardized technique, bile duct injury in SILC can be avoided.


[Remdesivir til behandling af COVID-19-pneumoni].


title: Persistent and stable gene expression by a cytoplasmic RNA replicon based on a noncytopathic variant Sendai virus.

text: Persistent and stable expression of foreign genes has been achieved in mammalian cells by integrating the genes into the host chromosomes. However, this approach has several shortcomings in practical applications. For example, large scale production of protein pharmaceutics frequently requires laborious amplification of the inserted genes to optimize the gene expression. The random chromosomal insertion of exogenous DNA also results occasionally in malignant transformation of normal tissue cells, raising safety concerns in medical applications. Here we report a novel cytoplasmic RNA replicon capable of expressing installed genes stably without chromosome insertion. This system is based on the RNA genome of a noncytopathic variant Sendai virus strain, Cl.151. We found that this variant virus establishes stable symbiosis with host cells by escaping from retinoic acid-inducible gene I-interferon regulatory factor 3-mediated antiviral machinery. Using a cloned genome cDNA of Sendai virus Cl.151, we developed a recombinant RNA installed with exogenous marker genes that was maintained stably in the cytoplasm as a high copy replicon (about 4 x 10(4) copies/cell) without interfering with normal cellular function. Strong expression of the marker genes persisted for more than 6 months in various types of cultured cells and for at least two months in rat colonic mucosa without any apparent side effects. This stable RNA replicon is a potentially valuable genetic platform for various biological applications.


COVID-19: Misinformation Can Kill.


title: Season of birth, clinical manifestations and Dexamethasone Suppression Test in unipolar major depression.

text: BACKGROUND Reports in the literature suggest that the season of birth might constitute a risk factor for the development of a major psychiatric disorder, possibly because of the effect environmental factors have during the second trimester of gestation. The aim of the current paper was to study the possible relationship of the season of birth and current clinical symptoms in unipolar major depression. METHODS The study sample included 45 DSM-IV major depressive patients and 90 matched controls. The SCAN v. 2.0, Hamilton Depression Rating Scale (HDRS) and Hamilton Anxiety Scale (HAS) were used to assess symptomatology, and the 1 mg Dexamethasone Suppression Test (DST) was used to subcategorize patients. RESULTS Depressed patients as a whole did not show differences in birth season from controls. However, those patients born during the spring manifested higher HDRS while those born during the summer manifested the lowest HAS scores. DST non-suppressors were almost exclusively (90%) likely to be born during autumn and winter. No effect from the season of birth was found concerning the current severity of suicidal ideation or attempts. DISCUSSION The current study is the first in this area of research using modern and rigid diagnostic methodology and a biological marker (DST) to categorize patients. Its disadvantages are the lack of data concerning DST in controls and a relatively small size of patient sample. The results confirm the effect of seasonality of birth on patients suffering from specific types of depression.


title: Modern metal-on-metal hip resurfacing.

text: For this technology overview, the tools of evidence-based medicine were used to summarize information on the indications, effectiveness, and failure rates of modern metal-on-metal hip resurfacing technology. The task was complicated by the fact that resurfacing arthroplasty is commonly offered only to a subset of patients who are candidates for total hip replacement, often prohibiting direct comparisons. Comprehensive literature searches were conducted to address four key questions addressing revision rates, patient characteristics, effectiveness of treatment, and whether improved technique, surgeon experience, and/or patient selection lead to improved outcomes. Despite data limitations, it is apparent that revision rates are higher after resurfacing than after total hip arthroplasty. Potential prognostic indicators did not yield a consistent predictor of patient-oriented outcomes (eg, pain relief) for either resurfacing arthroplasty or total hip replacement. Because of differences between patients who received hip resurfacing and those who received total hip arthroplasty, the results of studies comparing these techniques cannot be interpreted. Finally, changes in technique and increased experience result in a decrease in revision rates and femoral neck fractures and improved pain and hip scores in resurfacing.


A Clinical Lecture on Recent Advances in Abdominal Surgery.


title: Inhibition of HBV replication and gene expression in vitro and in vivo with a single AAV vector delivering two shRNA molecules.

text: Hepatitis B virus (HBV) infection is highly prevalent worldwide. The major challenge for current antiviral treatment is the elevated drug resistance that occurs via rapid viral mutagenesis. In this study, we developed AAV vectors to simultaneously deliver two or three shRNAs targeting different HBV-related genes. These vectors showed markedly better antiviral effects than ones that delivered a single shRNA in vitro. A dual shRNA expression vector (AAV-157i/1694i), which simultaneously expressed two shRNAs targeted the S and X genes of HBV, reduced HBsAg, HBeAg and HBV DNA levels by 87+/-4, 80.3+/-2.6 and 86.2+/- 7% respectively, eight days post-transduction. In a mouse model of prophylactic treatment, HBsAg and HBeAg were reduced to undetectable levels and the serum HBV DNA level was reduced by at least 100 fold. These results indicate that AAV-157i/1694i generates potent anti-HBV effects and that the strategy of constructing multi-shRNA expression vectors may lead to enhanced anti-HBV efficacy and overcome the evading mechanism of the virus and thus the development of drug resistance. [BMB reports 2009; 42(1): 59-64].


title: Molecular intermediates of fitness gain of an RNA virus: characterization of a mutant spectrum by biological and molecular cloning.

text: The mutant spectrum of a virus quasispecies in the process of fitness gain of a debilitated foot-and-mouth disease virus (FMDV) clone has been analysed. The mutant spectrum was characterized by nucleotide sequencing of three virus genomic regions (internal ribosome entry site; region between the two AUG initiation codons; VP1-coding region) from 70 biological clones (virus from individual plaques formed on BHK-21 cell monolayers) and 70 molecular clones (RT--PCR products cloned in E. coli). The biological and molecular clones provided statistically indistinguishable definitions of the mutant spectrum with regard to the distribution of mutations among the three genomic regions analysed and with regard to the types of mutations, mutational hot-spots and mutation frequencies. Therefore, the molecular cloning procedure employed provides a simple protocol for the characterization of mutant spectra of viruses that do not grow in cell culture. The number of mutations found repeated among the clones analysed was higher than expected from the mean mutation frequencies. Some components of the mutant spectrum reflected genomes that were dominant in the prior evolutionary history of the virus (previous passages), confirming the presence of memory genomes in virus quasispecies. Other components of the mutant spectrum were genomes that became dominant at a later stage of evolution, suggesting a predictive value of mutant spectrum analysis with regard to the outcome of virus evolution. The results underline the observation that greater insight into evolutionary processes of viruses may be gained from detailed clonal analyses of the mutant swarms at the sequence level.


H5N1 influenza and the implications for Europe.


title: Advancements in the battle against severe acute respiratory syndrome.

text: Severe acute respiratory syndrome (SARS) is a newly emerged infectious disease with a significant morbidity and mortality. The major clinical features include persistent fever, chills/rigor, myalgia, malaise, dry cough, headache and dyspnoea. Respiratory failure is the major complication of SARS and approximately 20% of patients may progress to acute respiratory distress syndrome requiring invasive mechanical ventilatory support. However, the severity is much milder in infected young children. Treatment of SARS was empirical in 2003 due to our limited understanding of this new disease. Protease inhibitors (lopinavir/ritonavir) in combination with ribavirin may play a role as antiviral therapy in the early phase, whereas the role of IFN and systemic steroid in preventing immune-mediated lung injury deserves further investigation. Knowledge of the genomic sequence of the SARS coronavirus has facilitated the development of rapid diagnostic tests. In addition, other antiviral treatment, RNA interference, monoclonal antibody, synthetic peptides, and vaccines are being developed. This paper provides a review of the epidemiology, clinical features and possible treatment strategies of SARS.


title: Chloroquine: novel uses & manifestations.

text: Chloroquine (CHQ) is a cheap, relatively well tolerated drug initially developed for the treatment of malaria in the 1930s. CHQ has, however, since accrued a plethora of uses in the treatment and amelioration of several other diseases and conditions because of its lysosomotropic properties. It also has characteristic physiological and systemic effects. This review gives an overview of the history and pharmacology of CHQ, and progresses to consider some of the mechanisms that may underlie its biochemical and physiological effects. Additionally, an overview of some of the novel uses of CHQ in the treatment of viral infections and cancer are presented. The antimalarial mechanisms of CHQ were not discussed in this review. The message is that CHQ, despite its welldocumented toxicity and adverse side effects may have important future uses that are associated with its lysosomotropic and immunomodulatory mechanisms. The possibility exists therefore that CHQ might be re-introduced into regular malaria treatment.


title: Protective lactogenic immunity conferred by an edible peptide vaccine to bovine rotavirus produced in transgenic plants.

text: Vaccines produced in transgenic plants constitute a promising alternative to conventional immunogens, presenting the possibility of stimulating secretory and systemic immunity against enteric pathogens when administered orally. Protection against enteric pathogens affecting newborn animals requires, in most cases, the stimulation of lactogenic immunity. Here, the group presents the development of an experimental immunogen based on expression of an immunorelevant peptide, eBRV4, of the VP4 protein of bovine rotavirus (BRV), which has been described as harbouring at least one neutralizing epitope as well as being responsible for the adsorption of the virus to epithelial cells. The eBRV4 epitope was efficiently expressed in transgenic alfalfa as a translational fusion protein with the highly stable reporter enzyme beta-glucuronidase (betaGUS), which served as a carrier, stabilized the synthesized peptide and facilitated screening for the higher expression levels in plants. Correlation of expression of the eBRV4 epitope in plants with those presenting the highest betaGUS activities was confirmed by a Western blot assay specific for the BRV peptide. The eBRV4 epitope expressed in plants was effective in inducing an anti-rotavirus antibody response in adult female mice when administered either intraperitoneally or orally and, more importantly, suckling mice born from immunized female mice were protected against oral challenge with virulent rotavirus. These results demonstrate the feasibility of inducing lactogenic immunity against an enteric pathogen using an edible vaccine produced in transgenic plants.


title: Percutaneous biliary stenting combined with radiotherapy as a treatment for unresectable hilar cholangiocarcinoma.

text: Hilar cholangiocarcinoma is often unresectable at the time of the initial diagnosis, and the provision of a definite palliative benefit is important in patients with unresectable hilar cholangiocarcinoma. The aim of the present study was to evaluate the safety of percutaneous biliary stenting and to analyze whether percutaneous biliary stenting combined with radiotherapy (RT) prolonged the stent patency and survival time of patients. In total, the cases of 38 patients with unresectable hilar cholangiocarcinoma that underwent percutaneous biliary stenting at the Navy General Hospital were retrospectively reviewed in the present study. Uncovered metallic stenting (UMS) combined with RT was administered to 25 patients, and UMS alone was administered to 13 patients. The records of early complications subsequent to percutaneous biliary stenting were collected, and the stent patency and survival times of patients were analyzed and compared between the two groups. The technical success rate of the procedure was 100% and the successful drainage rate was 86.8%. The overall early complication rate was 15.8% and the procedure-associated mortality rate was 2.6%. The median stent patency was 326 days in the UMS+RT group and 196 days in the UMS group (P=0.022). The UMS+RT group (median, 367 days) demonstrated a longer survival time compared with the UMS group (median, 267 days; P=0.025). Percutaneous biliary stenting offers a safe and effective method for the palliative treatment of patients with unresectable hilar cholangiocarcinoma, and percutaneous biliary stenting combined with RT may prolong stent patency and patient survival time.


title: Sertraline-related bleeding tendency: could it be dose-dependent?

text: Selective serotonin reuptake inhibitors (SSRIs) are reported to be associated with increased bleeding tendency. While findings of recent studies explain a lot about the pathophysiology of this side-effect, there is a general tendency to discontinue SSRIs as harmful medications. We report two instances of dose-dependent relations between sertraline and bleeding tendency. Bleeding diathesis was alleviated by adjusting dosage of medication. It could be argued that benefits of SSRIs could outweigh this potential and probably avoidable side-effect; if dose-adjustment is properly implemented.


title: Robotic First Rib Resection for Thoracic Outlet Syndrome.

text: PURPOSE First rib resection is a key component of the treatment of Thoracic Outlet Syndrome (TOS). We report our experience with, and technique for, robotic first rib resection. METHODS Patients diagnosed with TOS underwent robotic first rib resection of the offending portion of the first rib with disarticulation of the costo-sternal joint. Definitive diagnosis of TOS was made by Magnetic Resonance Angiography (MRA) with maneuvers. RESULTS A total of 67 patients underwent robotic first rib resection. Neurogenic TOS: 39 patients underwent robotic resection for Neurologic Symptoms of the upper extremity (Neurogenic TOS). There were 14 men and 25 women, with a mean age of 34 ± 9.5 years. Paget-Schroetter Syndrome (PSS) or Venous TOS: 28 patients underwent transthoracic robotic first rib resection for PSS. There were 16 men and 12 women, with a mean age of 24 ± 8.5 years. Operative time was 87.6 ± 10.8 minutes. There were no intraoperative complications. Hospital stay ranged from 2 to 4 days with a median hospitalization of 3 days. There were no neurovascular complications. There was no mortality. In patients with Neurogenic TOS, QuickDASH Scores (mean ± SEM) decreased from 60.3 ± 2.1 preoperatively to 5 ± 2.3 in the immediate postoperative period and to 3.5 ± 1.1 at 6 months (p<0001). Immediate relief of symptoms was seen in 35/39 patients (91%). Persistent paresthesia was seen in 4/39 (9%) immediately postop, and in 2/39 (2.5%) at 6 months. Thirty-seven of 39 (97.5%) patients reported complete relief of symptoms. Among patients with PSS or Venous TOS, 9/28 (32%) required endovascular venoplasty to completely open the subclavian vein after the relief of extrinsic compression. At a median follow-up of 24 months, all patients with PSS had an open subclavian vein, for a patency rate of 100%. CONCLUSIONS Robotic transthoracic first rib resection allows for minimally invasive resection of the first rib in patients with TOS, with excellent relief of symptoms and no neurovascular complications.


title: Prevalence of viral infections in cats in southwestern Poland in the years 2006 to 2010.

text: This study included data from 676 cats in southwestern Poland examined between 2006 and 2010. Examinations were performed to diagnose the following infections: feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), feline coronavirus (FCoV), feline calicivirus (FCV), and feline herpesvirus (FHV). The presence of antibodies or antigens was assessed in samples from all cats. Among examined cats, 46.9% were positive for FCV, 50% for FHV, and 38.5% for FCoV. Retroviral infections were detected more rarely: FeLV in 6.4% and FIV in 4.3% of cases.


title: Expression of the infectious bronchitis virus spike protein by recombinant vaccinia virus and induction of neutralizing antibodies in vaccinated mice.

text: A cDNA clone of the infectious bronchitis virus (IBV) spike protein gene has been recombined into vaccinia virus. Cells infected with the recombinant virus synthesized IBV spike antigen which was recognized by antibody raised against purified spike protein. Immunofluorescence showed that the IBV spike antigen was transported to the infected cell surface membrane and immunoprecipitation showed the presence of the glycosylated 180K mol. wt. polypeptide precursor of the two spike subunits S1 and S2 that comigrated with this antigen from IBV-infected cells. Vaccinated mice produced antibody that recognized the IBV spike antigen by ELISA and which neutralized IBV infectivity as shown by ciliostasis tests on tracheal organ cultures.


Bovine Tuberculosis.


title: Severe acute respiratory distress syndrome (SARS): a critical care perspective.

text: OBJECTIVE To review the epidemiology, clinical features, etiology, diagnosis, and management of severe acute respiratory syndrome (SARS) from a critical care perspective. DATA SOURCES A MEDLINE search was performed using the following terms: severe acute respiratory syndrome and SARS virus. Additional information and references were obtained from the Web sites for the Centers for Disease Control and Prevention, World Health Organization, and Health Canada. STUDY SELECTION Recent case series were used to develop a review of the epidemiology, clinical features, outcomes, and management of patients with SARS from an intensive care unit (ICU) perspective. This was supplemented by epidemiology information obtained from other Web-based sources. Recent published studies describing the etiology of SARS were also included. DATA SYNTHESIS SARS has rapidly spread from Southeast Asia to numerous countries, including Canada and the United States. A new coronavirus has been isolated and detected from many affected patients. The mortality rate worldwide is approximately 10.5%. From five cohorts, the ICU admission rate ranged from 20% to 38%. Fifty-nine percent to 100% of the ICU patients required mechanical ventilatory support. The mortality rate of SARS patients admitted to the ICU ranged from 5% to 67%. The most common clinical symptoms and signs are fever, cough, dyspnea, myalgias, malaise, and inspiratory crackles. Common laboratory abnormalities included mild leukopenia, lymphopenia, and increased aspartate transaminase, alanine transaminase, lactic dehydrogenase, and creatine kinase. The chest radiograph pattern ranged from focal infiltrates to diffuse airspace disease. Management consisted of isolation, strict respiratory and contact precautions, ventilatory support as needed, empiric broad-spectrum antibiotics, ribavirin, and corticosteroids. Predictors of mortality included advanced age, the presence of comorbidities, and a high lactic dehydrogenase or high neutrophil count at admission. CONCLUSIONS SARS is a highly contagious, infectious process that can advance to significant hypoxemic respiratory failure requiring ICU monitoring and support. Early recognition is critical for effective management and containment of this disease.


Pediatric Allergy Asthma Meeting - PAAM, Berlin, October 2015.


title: Transgenic mice expressing a soluble form of porcine nectin-1/herpesvirus entry mediator C as a model for pseudorabies-resistant livestock.

text: An approach to genetically engineered resistance to pseudorabies virus (PRV) infection was examined by using a transgene encoding a soluble form of nectin-1, also known as herpesvirus entry mediator C. Nectin-1 is an alpha-herpesvirus receptor that binds to virion glycoprotein D. Nectin-1 mediates entry of PRV, herpes simplex virus types 1 and 2, and bovine herpesvirus type 1. To assess the antiviral potential of an ectopic expression of the nectin-1 ectodomain in vivo, six transgenic mouse lines expressing a soluble form of nectin-1, consisting of an extracellular domain of porcine nectin-1 and the Fc portion of human IgG1, were generated. All of the transgenic mouse lines showed nearly complete resistance to PRV infection by means of both i.p. and intranasal routes. These results suggest that the introduction into farm animals of a transgene encoding a soluble form of nectin-1 would offer a potent biological approach to generating alpha-herpesvirus-resistant livestock.


Looking down is looking up.


title: Subcellular localization and membrane topology of the Dengue virus type 2 Non-structural protein 4B.

text: Dengue virus (DV) is a member of the family Flaviviridae. These positive strand RNA viruses encode a polyprotein that is processed in case of DV into 10 proteins. Although for most of these proteins distinct functions have been defined, this is less clear for the highly hydrophobic non-structural protein (NS) 4B. Despite its possible role as an antagonist of the interferon-induced antiviral response, this protein may play an additional more direct role for viral replication. In this study we determined the subcellular localization, membrane association, and membrane topology of DV NS4B. We found that NS4B resides primarily in cytoplasmic foci originating from the endoplasmic reticulum. NS4B colocalizes with NS3 and double-stranded RNA, an intermediate of viral replication, arguing that NS4B is part of the membrane-bound viral replication complex. Biochemical analysis revealed that NS4B is an integral membrane protein, and that its preceding 2K signal sequence is not required for this integration. We identified three membrane-spanning segments in the COOH-terminal part of NS4B that are sufficient to target a cytosolic marker protein to intracellular membranes. Furthermore, we established a membrane topology model of NS4B in which the NH2-terminal part of the protein is localized in the endoplasmic reticulum lumen, whereas the COOH-terminal part is composed of three trans-membrane domains with the COOH-terminal tail localized in the cytoplasm. This topology model provides a good starting point for a detailed investigation of the function of NS4B in the DV life cycle.


title: Severe acute respiratory syndrome: scientific and anecdotal evidence for drug treatment.

text: Severe acute respiratory syndrome (SARS), caused by a highly infectious novel coronavirus (CoV), predominantly presents with severe pneumonitis leading to respiratory failure and death in approximately 10% of victims. Most cases present, after an incubation of 2 to 11 days, with fever and chills, which are followed by dry cough and dyspnea before the onset of respiratory failure. The management of SARS is controversial, largely due to the lack of data from controlled trials, which were logistically impossible to design or execute at the time of the overwhelming outbreak between March and June 2003. The use of an antiviral is logical although there is no effective agent against SARS-CoV, with the widespread use of ribavirin in 2003 attracting considerable scepticism. The use of ribavirin as a monotherapy in SARS is not recommended. Retrospective data suggest that administration of the anti-HIV drug Kaletra in combination with ribavirin could reduce mortality and incidence of respiratory failure. The use of corticosteroid was based on the similarity between SARS and bronchiolitis obliterans organizing pneumonia. Corticosteroid use is considered important in subsets of patients with SARS. As SARS may still resurge, it is imperative that past experience is analyzed. This review will attempt to address the rationale for pharmacotherapy in SARS using anecdotal and the limited published data.


title: Preferential infection of mature dendritic cells by mouse hepatitis virus strain JHM.

text: Mouse hepatitis virus strain JHM (MHV-JHM) causes acute encephalitis and acute and chronic demyelinating diseases in mice. Dendritic cells (DCs) are key cells in the initiation of innate and adaptive immune responses, and infection of these cells could potentially contribute to a dysregulated immune response; consistent with this, recent results suggest that DCs are readily infected by another strain of mouse hepatitis virus, the A59 strain (MHV-A59). Herein, we show that the JHM strain also productively infected DCs. Moreover, mature DCs were at least 10 times more susceptible than immature DCs to infection with MHV-JHM. DC function was impaired after MHV-JHM infection, resulting in decreased stimulation of CD8 T cells in vitro. Preferential infection of mature DCs was not due to differential expression of the MHV-JHM receptor CEACAM-1a on mature or immature cells or to differences in apoptosis. Although we could not detect infected DCs in vivo, both CD8(+) and CD11b(+) splenic DCs were susceptible to infection with MHV-JHM directly ex vivo. This preferential infection of mature DCs may inhibit the development of an efficient immune response to the virus.


title: Diagnosis and management of acute bronchitis.

text: Acute bronchitis is one of the top 10 conditions for which patients seek medical care. Physicians show considerable variability in describing the signs and symptoms necessary to its diagnosis. Because acute bronchitis most often has a viral cause, symptomatic treatment with protussives, antitussives, or bronchodilators is appropriate. However, studies indicate that many physicians treat bronchitis with antibiotics. These drugs have generally been shown to be ineffective in patients with uncomplicated acute bronchitis. Furthermore, antibiotics often have detrimental side effects, and their overuse contributes to the increasing problem of antibiotic resistance. Patient satisfaction with the treatment of acute bronchitis is related to the quality of the physician-patient interaction rather than to prescription of an antibiotic.


[Unique mechanism of coronavirus mRNA transcription].


title: [Gene detection of severe acute respiratory syndrome-related coronavirus].

text: OBJECTIVE To develop a newly real-time RT-polymerase chain reaction assay for severe acute respiratory syndrome (SARS) related coronavirus in human whole blood. METHODS A pair of primers and a probe (molecular beacon) had been designed that were specific for the recognition of a highly conservative region between 15 301 and 15 480 of the SARS-related coronavirus polymerase gene sequences obtained from GenBank (G130027616). RESULTS In the real-time RT-PCR assay, the extent of SARS related coronavirus amplification was measured in terms of the increase in fluorescence during the amplification process. The 145 bp fragment of PCR product was further confirmed by conventional PCR assay and proved by DNA sequencing to be identical to the target sequence to which the probe was hybridized. CONCLUSION This assay has a broad application for clinical diagnosis and surveillance investigation.


title: Eliciting Disease Data from Wikipedia Articles.

text: Traditional disease surveillance systems suffer from several disadvantages, including reporting lags and antiquated technology, that have caused a movement towards internet-based disease surveillance systems. Internet systems are particularly attractive for disease outbreaks because they can provide data in near real-time and can be verified by individuals around the globe. However, most existing systems have focused on disease monitoring and do not provide a data repository for policy makers or researchers. In order to fill this gap, we analyzed Wikipedia article content. We demonstrate how a named-entity recognizer can be trained to tag case counts, death counts, and hospitalization counts in the article narrative that achieves an F1 score of 0.753. We also show, using the 2014 West African Ebola virus disease epidemic article as a case study, that there are detailed time series data that are consistently updated that closely align with ground truth data. We argue that Wikipedia can be used to create the first community-driven open-source emerging disease detection, monitoring, and repository system.


title: Mining human antibody repertoires.

text: Human monoclonal antibodies (mAbs) have become drugs of choice for the management of an increasing number of human diseases. Human antibody repertoires provide a rich source for human mAbs. Here we review the characteristics of natural and non-natural human antibody repertoires and their mining with non-combinatorial and combinatorial strategies. In particular, we discuss the selection of human mAbs from naïve, immune, transgenic, and synthetic human antibody repertoires using methods based on hybridoma technology, clonal expansion of peripheral B cells, single-cell PCR, phage display, yeast display, and mammalian cell display. Our reliance on different strategies is shifting as we gain experience and refine methods to the efficient generation of human mAbs with superior pharmacokinetic and pharmacodynamic properties.


title: Detection of infectious bronchitis virus by real-time reverse transcriptase-polymerase chain reaction and identification of a quasispecies in the Beaudette strain.

text: In this report, we describe a real-time reverse transcriptase-polymerase chain reaction (RRT-PCR) diagnostic test for infectious bronchitis virus (IBV) with the use of fluorescence resonance energy transfer (FRET) technology. Two primers that amplify a 383-base pair product between nucleotide positions 703 and 1086 relative to the start codon for the S1 gene of the Massachusetts 41 virus were designed and used to amplify the Beaudette, Massachusetts 41, Florida 18288, Connecticut, Iowa 97, Arkansas DPI, CA/NE95/99, DE/072/ 92, and GA/0470/98 strains of IBV. The primers were specific and did not amplify New Castle disease virus, Mycoplasma spp., or infectious laryngotracheitis virus. For RRT-PCR by FRET, an anchor probe conjugated to fluorescein and a detection probe conjugated to a red fluorophore were designed to anneal to a hypervariable region within the 383-base pair product. The level of sensitivity was 1 x 10(4) RNA molecules used as starting template. After amplification, a melting curve analysis was conducted to specifically identify IBV types. Because of sequence differences in the annealing position of the detection probe, the Arkansas, Connecticut, Beaudette, and Massachusetts 41 strains could be differentiated. No fluorescence was observed for the DE/072/ 92 and GA/0470/98 viruses with the anchor and detection probes. When the Beaudette strain was examined, two melting peaks were observed at 44 C and 51 C, indicating a quasispecies in that laboratory strain of IBV. Routine typing of vaccine strains of IBV was possible with this technology, but high standard deviations associated with the melting curve analysis of the FRET probes described herein made it difficult to use this test reliably for routine typing of IBV field isolates.


title: Innovative technology conserves resources and generates savings: a case study from the Sunnybrook Regional Processing Centre.

text: The regional processing centre at Sunnybrook Health Sciences Centre recently faced the substantial challenge of increasing cleaning capacity to meet the current workload and anticipated future demand without increasing its operating budget. The solution, upgrading its cleaning and decontamination system to a highly automated system, met both objectives. An analysis of the impact of the change found that the new system provided additional benefits, including improved productivity and cleaning quality; decreased costs; reduced water, electricity and chemical use; improved worker safety and morale; and decreased overtime. Investing in innovative technology improved key departmental outcomes while meeting institutional environmental and cost savings objectives.


title: Insulin pump therapy for 1-6 year old children with type 1 diabetes.

text: BACKGROUND The management of diabetes in preschool children poses unique difficulties for both the families and the medical team. OBJECTIVE To test the feasibility and safety of insulin pump therapy in the 1-6 year age group in order to improve quality of life and metabolic control. METHODS The study group comprised 15 type 1 diabetic children aged 1-6 years old (mean +/- SD, 3.8 +/- 1.2 years) from three diabetes centers. Insulin pump therapy was applied for 12 months. Data, including insulin dose, hemoglobin A1c, hypoglycemic events, as well as scores on the Diabetes Quality of Life Measure Questionnaire and the Diabetes Treatment Satisfaction Questionnaire, were collected and compared with the multiple daily injection treatment prior to entry into the study, RESULTS HbA1c was measured at the beginning of the study and at 2, 4, 8 and 12 months later; the respective levels (mean +/- SD) were 8.82 +/- 0.98, 8.45 +/- 1.05, 8.37 +/- 0.85, 8.32 +/- 0.71, 8.18 +/- 0.90%. HbA1c measurements after 12 months were significantly lower than at the beginning of the study (P < 0.05). There were no significant differences in insulin dose and the total number of hypoglycemic events. In both the DQOL and DTSQ scales there were significant differences in scores in favor of the insulin pump period (43.7 +/- 8.0 versus 33.7 +/- 7.9, P < 0.001; and 10.9 +/- 2.3 versus 14.5 +/- 2.3, P < 0.001), respectively. CONCLUSIONS For very young diabetic children, insulin pump therapy improves quality of life and is feasible and safe. It should be considered as an optional mode of therapy for this age group.


Infectious diseases. SARS experts in China urge wider animal testing.


Multiple rpoB mutants of Mycobacterium tuberculosis and second-order selection.


title: Public health response to two incidents of confirmed MERS-CoV cases travelling on flights through London Heathrow Airport in 2014 – lessons learnt.

text: In May 2014, Public Health England was alerted to two separate laboratory-confirmed cases of Middle East respiratory syndrome coronavirus (MERS-CoV) infection who transited through London Heathrow Airport while symptomatic on flights from Saudi Arabia to the United States of America. We present the rationale for the public health response to both incidents, and report results of contact tracing. Following a risk assessment, passengers seated two seats around the cases were prioritised for contact tracing and a proactive media approach was used to alert all passengers on the planes of their possible exposure in both incidents. In total, 64 United Kingdom (UK) residents were successfully contacted, 14 of whom were sat in the priority area two seats all around the case(s). Five passengers reported respiratory symptoms within 14 days of the flight, but all tested were negative for MERS-CoV. Details of non-UK residents were passed on to relevant World Health Organization International Health Regulation focal points for follow-up, and no further cases were reported back. Different approaches were used to manage contact tracing for each flight due to variations in the quality and timeliness of the passenger contact information provided by the airlines involved. No evidence of symptomatic onward transmission was found.


title: Prevention of virus persistence and protection against immunopathology after Borna disease virus infection of the brain by a novel Orf virus recombinant.

text: The Parapoxvirus Orf virus represents a promising candidate for novel vector vaccines due to its immune modulating properties even in nonpermissive hosts such as mouse or rat. The highly attenuated Orf virus strain D1701 was used to generate a recombinant virus (D1701-VrVp40) expressing nucleoprotein p40 of Borna disease virus, which represents a major antigen for the induction of a Borna disease virus-specific humoral and cellular immune response. Infection with Borna disease virus leads to distinct neurological symptoms mediated by the invasion of activated specific CD8+ T cells into the infected brain. Usually, Borna disease virus is not cleared from the brain but rather persists in neural cells. In the present study we show for the first time that intramuscular application of the D1701-VrVp40 recombinant protected rats against Borna disease, and importantly, virus clearance from the infected brain was demonstrated in immunized animals. Even 4 and 8 months after the last immunization, all immunized animals were still protected against the disease. Initial characterization of the immune cells attracted to the infected brain areas suggested that D1701-VrVp40 mediated induction of B cells and antibody-producing plasma cells as well as T cells. These findings suggest the induction of various defense mechanisms against Borna disease virus. First studies on the role of antiviral cytokines indicated that D1701-VrVp40 immunization did not lead to an enhanced early response of gamma or alpha interferon or tumor necrosis factor alpha. Collectively, this study describes the potential of the Orf virus vector system in mediating long-lasting, protective antiviral immunity and eliminating this persistent virus infection without provoking massive neuronal damage.


title: Role of law enforcement response and microbial forensics in investigation of bioterrorism.

text: The risk and threat of bioterrorism and biocrime have become a large concern and challenge for governments and society to enhance biosecurity. Law enforcement plays an important role in assessing and investigating activities involved in an event of bioterrorism or biocrime. Key to a successful biosecurity program is increased awareness and early detection of threats facilitated by an integrated network of responsibilities and capabilities from government, academic, private, and public assets. To support an investigation, microbial forensic sciences are employed to analyze and characterize forensic evidence with the goal of attribution or crime scene reconstruction. Two different molecular biology-based assays--real time polymerase chain reaction (PCR) and repetitive element PCR--are described and demonstrate how molecular biology tools may be utilized to aid in the investigative process. Technologies relied on by microbial forensic scientists need to be properly validated so that the methods used are understood and so that interpretation of results is carried out within the limitations of the assays. The three types of validation are preliminary, developmental, and internal. The first is necessary for rapid response when a threat is imminent or an attack has recently occurred. The latter two apply to implementation of routinely used procedures.


title: Emerging waterborne infections: contributing factors, agents, and detection tools.

text: Because microorganisms are easily dispersed, display physiological diversity, and tolerate extreme conditions, they are ubiquitous and may contaminate and grow in water. The presence of waterborne enteric pathogens (bacteria, viruses, and protozoa) in domestic water supplies represents a potentially significant human health risk. Even though major outbreaks of waterborne disease are comparatively rare, there is substantial evidence that human enteric pathogens that are frequently present in domestic water supplies are responsible for low-level incidence of waterborne microbial disease. Although these diseases are rarely debilitating to healthy adults for more than a few hours to a few days, enteric pathogens can cause severe illness, even death, for young children, the elderly, or those with compromised immune systems. As the epidemiology of waterborne diseases is changing, there is a growing global public health concern about new and reemerging infectious diseases that are occurring through a complex interaction of social, economic, evolutionary, and ecological factors. New microbial pathogens have emerged, and some have spread worldwide. Alternative testing strategies for waterborne diseases should significantly improve the ability to detect and control the causative pathogenic agents. In this article, we provide an overview of the current state of knowledge of waterborne microbial pathogens, their detection, and the future of new methods in controlling these infectious agents.


Background paper. Functions of the coronavirus nucleocapsid protein.


National Notifiable Diseases Surveillance System tables.


Dangers of postoperative opioids: is there a cure?


title: Association study of ACE2 (angiotensin I-converting enzyme 2) gene polymorphisms with coronary heart disease and myocardial infarction in a Chinese Han population.

text: Results are accumulating that ACE2 (angiotensin I-converting enzyme 2) might act as a protective protein for cardiovascular diseases; however, only a few studies in human populations have been carried out. This prompted us to perform a case-control study to investigate the relationship of ACE2 polymorphisms with CHD (coronary heart disease) and MI (myocardial infarction). Three single nucleotide polymorphisms in the ACE2 gene (1075A/G, 8790A/G and 16854G/C) were genotyped by PCR-RFLP (restriction-fragment-length polymorphism) in 811 patients with CHD (of which 508 were patients with MI) and 905 normal controls in a Chinese population. The polymorphisms were in linkage disequilibrium (r(2)=0.854-0.973). Analyses were conducted by gender, because the ACE2 gene is on the X chromosome. In females, an association was detected with MI for 1075A/G (P=0.026; odds ratio=1.98) and 16854G/C (P=0.028; odds ratio=1.97) in recessive models after adjusting for covariates. In male subjects, two haplotypes (AAG and GGC) were common in frequency. In male subjects not consuming alcohol, the haplotype GGC was associated with a 1.76-fold risk of CHD [95% CI (confidence interval), 1.15-2.69; P=0.007] and a 1.77-fold risk of MI (95% CI, 1.12-2.81; P=0.015) with environmental factors adjusted, when compared with the most common haplotype AAG. In conclusion, the results of the present study indicate that common genetic variants in the ACE2 gene might impact on MI in females, and may possibly interact with alcohol consumption to affect the risk of CHD and MI in Chinese males.


title: General practice: professional preparation for a pandemic.

text: General practice will play a key role in both prevention and management of an influenza pandemic. Australian pandemic plans acknowledge a role for general practice, but there are few published data addressing the issues that general practitioners and their practices will face in dealing with such a crisis. The outcome will revolve around preparation in three key areas: Definition of the role of general practice within a broad primary care pandemic response, and adequate preparation within general practices so they can play that role well. Planning exercises and forums must include GPs, and rehearsals must include practical experience for general practices and their staff. Local Divisions of General Practice and GP practices can advocate for this, can define their role, and can prepare by using pandemic preparedness checklists; Definition and enactment of communication strategies to facilitate transfer of useful clinical and administrative data from practices and rapid dissemination of information into the community via general practice; Resource provision, which should be centrally funded but locally distributed, with personal protective equipment, vaccines and antivirals readily available for distribution. Resources must include support for human resource management to ensure appropriate health care professionals reach areas of workforce demand. Administrative, clinical and financial resources must be available to train GPs and practices in pandemic awareness and response.


Travel by air: health considerations.


title: Genetic variation in porcine reproductive and respiratory syndrome virus isolates in the midwestern United States.

text: The nucleotide sequence of a 3266 bp region encompassing open reading frames (ORFs) 2 through 7 of the porcine reproductive and respiratory syndrome virus (PRRSV) was determined for 10 isolates recovered from the midwestern United States. Pairwise comparisons showed that genetic distances between isolates ranged from 2.5% to 7.9% (mean 5.8% +/- 0.2%) whereas the Lelystad strain from Europe was, on average, 34.8% divergent from US clones. Thus, US and European PRRSV isolates represent genetically distinct clusters of the same virus. ORF 5, which encodes the envelope glycoprotein, was the most polymorphic [total nucleotide diversity (pi) = 0.097 +/- 0.007] and ORF 6, encoding the viral M protein, was the most conserved (pi = 0.038 +/- 0.003). The substantial differences in nucleotide diversity among ORFs suggests that the virus is evolving by processes other than simple accumulation of random neutral mutations. In support of this hypothesis, statistical analyses of the nucleotide sequence provided strong evidence for intragenic recombination or gene conversion in ORFs 2, 3, 4, 5 and 7, but not in ORF 6. An excess of synonymous (silent) substitutions was observed in all six ORFs, indicating an evolutionary pressure to conserve amino acid sequences. Taken together, the data indicate that despite intragenic recombination among extant PRRSV isolates, purifying selection has acted to maintain the primary structure of individual ORFs.


title: Contribution of passive immunity to porcine respiratory coronavirus to protection against transmissible gastroenteritis virus challenge exposure in suckling pigs.

text: OBJECTIVE To determine the ability of porcine respiratory coronavirus (PRCV) infections to induce passive immunity in suckling pigs to transmissible gastroenteritis virus (TGEV) challenge exposure. DESIGN AND ANIMALS: 4 TGEV seronegative sows and their litters (group A) served as controls, whereas 2 other groups (B and C) of sows (also TGEV seronegative) were oronasally inoculated with live PRCV during 1 or 2 subsequent pregnancies, respectively. PROCEDURE Effectiveness of passive immunity provided to pigs via colostrum and milk was assessed after TGEV challenge exposure, and TGEV antibody responses in colostrum and milk were analyzed. RESULTS Mortality in the 3 groups of young pigs correlated with severity of clinical signs of TGEV infection and was highest in control litters (86% in group-A pigs) and lowest in litters of sows inoculated with PRCV in 2 subsequent pregnancies (14% in group-C pigs). Virus-neutralization and IgA and IgG TGEV antibody titers of milk collected from sows at challenge exposure had significant positive correlation with litter survival. Significantly higher numbers of TGEV-specific IgA and IgG antibody-secreting cells were found in group-A pigs than in group-C pigs, suggesting that high titer of maternal antibodies (induced in group-C sows multiply exposed to PRCV) may interfere with active antibody responses. CONCLUSIONS AND CLINICAL RELEVANCE Results suggest that, in PRCV-infected pig herds, multiple exposures of pregnant sows are associated with higher IgA and IgG antibody titers to TGEV in milk, and these titers contribute to protection against TGEV infection.


title: Depression, anxiety, body image, sexual functioning, and dyadic adjustment associated with dialysis type in chronic renal failure.

text: OBJECTIVE Depression is the most widely studied complication in dialysis patients. In patients with chronic renal failure, changes in body image are considered to be associated with invasive treatment interventions. In addition, sexual problems are common in dialysis patients. In this study, hemodialysis and peritoneal dialysis patients are investigated for depression, anxiety, body image, sexual satisfaction, and dyadic adjustment. METHODS Hemodialysis patients (n = 36), peritoneal dialysis patients (n = 54), and healthy controls (n = 30) were included in the study. All the subjects were assessed with Structured Clinical Interview for DSM-IV, Body Image Scale, Beck Depression Inventory, Beck Anxiety Scale, Golombok-Rust Inventory for Sexual Satisfaction, and Dyadic Adjustment Scale. RESULTS Depression (20.64 +/- 15.20) and anxiety levels (14.72 +/- 12.36) were significantly higher in hemodialysis group compared to peritoneal dialysis (13.54 +/- 12.51; 12.74 +/- 11.21) and control groups (7.17 +/- 5.58; 9.86 +/- 9.19). In peritoneal dialysis group, as depression and anxiety levels increased, body image was disturbed and sexual satisfaction decreased. In peritoneal dialysis group, body image (86.98 +/- 23.63) was better than hemodialysis group (101.58 +/- 26.51) and was not different from the control group (83.67 +/- 22.11). In hemodialysis group, as depression and anxiety levels increased, body image was disturbed. In both groups, long-term dialysis disturbed body image. CONCLUSION Patients should be informed about the impacts of dialysis. Clinicians may wish to monitor dialysis-users for anxiety, depression, dyadic adjustment, and body image difficulties at follow-up appointments. Interventions that target intimate partner interventions, appearance-related beliefs, and anxiety depression may be of benefit to this population.


title: Histological changes in intestine in semichronic diarrhoea induced by lactose enriched diet in rats: effect of Diarex-Vet.

text: Efficacy of Diarex-Vet (The Himalaya Drug Company, Makali, Bangalore, India) was evaluated histologically in semichronic diarrhoea induced by lactose enriched diet in rats. The rats in different groups were given lactose enriched diet alone for 2 days before starting the treatment with Diarex-Vet at a dose of 250, 500 and 750 mg/kg body weight along with lactose enriched diet for 5 days. Animals were euthanised at the end of 5 days of treatment and histological changes were observed in the ileum, caecum and colon. Semiquantitative analysis of goblet cells in intestines showed dose dependent response among the treated groups. The morphological changes were comparable to normal in the group given 750 mg/kg body wt Diarex-Vet. The effects observed were attributed to the lactase like analogous activity of Diarex-Vet or the inhibition of the osmotic processes in the intestinal lumen thereby reducing the morphological changes in the intestines.


title: Correlation of quantitative dynamic contrast-enhanced MRI with microvascular density in necrotic, partial necrotic, and viable liver tumors in a rabbit model.

text: The purpose of this study was to examine the correlation of quantitative dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) with microvessel density (MVD) in necrotic, partial necrotic, and viable tumors using a rabbit VX2 liver tumor model. Nine rabbits were used for this study. The complete necrotic area (CNA), partial necrotic area (PNA), and viable tumor area (VTA) of liver tumors were experimentally induced by radiofrequency ablation (RFA). DCE-MRI data were processed based on the extended Kety model to estimate Ktrans, ve and vp parameters. The boundaries among CNA, PNA, and VTA were delineated based on H&E stain images, and MVD was assessed for each subregion of each VX2 tumor based. There were no correlations between ph-parameters (Ktrans, ve, and vp) and MVD for CNA. For PNA, the Ktrans values were positively correlated with the MVD (r = 0.8124, p < 0.001). For VTA, we found a positive correlation between Ktrans values and the MVD (r = 0.5743, p < 0.05). Measuring from both the PNA and the VTA, mean Ktrans values were positively correlated with mean MVD (r = 0.8470, p < 0.0001). In a rabbit VX2 liver tumor model, Ktrans values correlated well with MVD counts of PNA and VTA in liver tumors.


title: Malaysia Stroke Council guide on acute stroke care service during COVID-19 Pandemic.

text: On the 18th of March 2020, the Malaysia government declared a movement control order (MCO) due to the unprecedented COVID-19 pandemic. Although the majority of patients presented with respiratory-related symptoms, COVID-19 patients may present atypically with neurological manifestations and may even have an increased risk of stroke. The Malaysia Stroke Council is concerned regarding the level of care given to stroke patients during this pandemic. During the recent National Stroke Workflow Steering Committee meeting, a guide was made based on the currently available evidences to assist Malaysian physicians providing acute stroke care in the hospital setting in order to provide the best stroke care while maintaining their own safety. The guide comprises of prehospital stroke awareness, hyperacute stroke care, stroke care unit and intensive care unit admission, post-stroke rehabilitation and secondary prevention practice. We urge continuous initiative to provide the best stroke care possible and ensure adequate safety for both patients and the stroke care team.


title: Prokaryote phylogeny without sequence alignment: from avoidance signature to composition distance.

text: This is a review of a new and essentially simple method of inferring phylogenetic relationships from complete genome data without using sequence alignment. The method is based on counting the appearance frequency of oligopeptides of a fixed length (up to K = 6) in the collection of protein sequences of a species. It is a method without fine adjustment and choice of genes. Applied to prokaryotic genomes it has led to results comparable with the bacteriologists' systematics as reflected in the latest 2002 outline of the Bergey's Manual of Systematic Bacteriology. The method has also been used to compare chloroplast genomes and to the phylogeny of Coronaviruses including human SARS-CoV. A key point in our approach is subtraction of a random background from the original counts by using a Markov model of order K-2 in order to highlight the shaping role of natural selection. The implications of the subtraction procedure is specially analyzed and further development of the new approach is indicated.


World no-tobacco day.


title: [Study on the changing regularity of special antibody and expression of stomach and enteric involvement on SARS-coronavirus infection in the recovery period of severe acute respiratory syndrome].

text: OBJECTIVE To study the change of special antibodies titer IgG, IgM and nucleocapsid to SARS coronavirus (CoV) and observing the expression of stomach and enteric involvement on SARS-CoV infection by monoclonal antibody against N protein of SARS-CoV in the 7-year recovery period among family clustering cases of severe acute respiratory syndrome. METHODS Special antibody titer to SARS-CoV of 14 patients from 5 different families and their 10 kinfolks continuously tested by IFA and antigen-capturing ELISA methods. Samples were taken in the 1(st) - 7(th) year periods after SARS patients infected by SARS-CoV, being diluted and measured on it titers of three kinds of antibodies. Immunochemical staining with monoclonal antibody (mAb) against N protein of SARS-CoV was used to determine the stomach and enteric tissues among 5 SARS patients with their nucleocapsid antibody titer ascended obviously after 1(st)-7(th) year. RESULTS When testing the IgG antibody titer of the 14 SARS patients by IFA method, the average titer was 1/71 (95%CI: 1/58 - 1/85) in the 1(st) year, but began to descend in the following years, and the IgG antibody of the most SARS patients disappeared in the 7(th) year. Regarding the IgM titer, it disappeared in most of the SARS patients 1 year later. The average value of nucleocapsid antibody titer was 1/146 (95%CI: 1/122 - 1/171) in the 1(st) year, and it descended as the IgG antibody titer did. In 5 cases, differences appeared. The nucleocapsid antibody titer was between 1/156 and 1/210 in 3 cases, and 2 cases were normal. Immunochemical staining with mAb against N protein of SARS-CoV was identified in the stomach and enteric tissues of 5 SARS patients with the nucleocapsid antibody titer increased significantly, 1(st)-7(th) year later. The five patients were detected by gastroscopy detection and cell immunohistochemistry test. 3 cases showed N protein antibody positive in the serum, and positive immunohistochemical expression in most of the cytoplasm in the gastric tissue mucous gland epithelial cells. 1 case also expressed in the intestinal tissue slurry columnar epithelium and interstitial cells. The other two cases showed negative on both serum N protein antibody and immunohistochemical expression. The biopsy results of the 5 patients were as follows: 1 case diagnosed as "signet-ring cell carcinoma of the stomach and rectum multiple transfer", 1 case of gastric polyp, 1 case of superficial antral gastritis and 2 cases were normal. CONCLUSION By testing the special IgG, IgM, nucleocapsid antibody to SARS-CoV of the 14 family clustering cases, we found that they all decreased in the 7(th) year, and most of them disappeared. The nucleocapsid antibody titer was related to pathogenetic condition. SARS-CoV was proved to be still present in stomach and enteric tissues of SARS patients with the nucleocapsid antibody titer increased significantly after the 7(th) year.


title: Comparative analysis of techniques to prevent laparoscopic fogging.

text: PURPOSE To evaluate the efficiency and efficacy of two common and commercially available methods to resolve lens fogging: a Fluid Warming System (O.R. Solutions, Chantilly, VA) and the Clearify Visualization System (Covidien, Mansfield, MA). MATERIAL AND METHODS We prospectively evaluated 40 patients undergoing laparoscopic renal procedures with the Fluid Warming System (first 20 cases) and then the Clearify Visualization System (second 20 cases). We utilized the standard Fluid Warming System per a 30-second modified protocol established in our laboratory. We used the Clearify according to manufacturer instructions. For each procedure we documented the etiology of each episode of visual obstruction, procedure type and surgery duration. We performed a cost analysis. For all cases we used the same insufflator, insufflation trocar location and trocar configuration. RESULTS All 40 patients completed the study protocol without incident. The mean fogging events per hour for the Fluid Warming System and Clearify Visualization System were 0.7 (0-2.52) and 1.4 (0-5.02), respectively (P = 0.045). Surgery duration and cost per procedure were similar for both systems. CONCLUSION The Fluid Warming System with modified technique was found to have less fogging events than the Clearify Visualization System, with no difference in operative time or cost.


title: Mild dilatation of renal pelvis in term neonates with urinary tract infection.

text: The purpose of this study was to assess the incidence and evolution of mild dilatation confined to the renal pelvis in term neonates with urinary tract infection developing within 2 weeks from birth. Twenty-two neonates with mild dilatation of the renal pelvis out of 180 neonates with urinary tract infection were identified giving an incidence of 12.2% for this finding. Male to female ratio was 6.3:1. The left kidney was twice as frequently involved (68 vs. 32%). At follow-up, the dilatation had disappeared in 20 neonates (90.9%) by a mean age of 2.7 years with only one neonate developing two further episodes of urinary tract infection in the infantile period. No other morbidity was noted.


title: Severe acute respiratory syndrome coronavirus Orf3a protein interacts with caveolin.

text: The orf3a (also called X1 or U274) gene is the largest unique open reading frame in the severe acute respiratory syndrome coronavirus genome and has been proposed to encode a protein with three transmembrane domains and a large cytoplasmic domain. Recent work has suggested that the 3a protein may play a structural role in the viral life cycle, although the mechanisms for this remain uncharacterized. Here, the expression of the 3a protein in various in vitro systems is shown, it has been localized to the Golgi region and its membrane topology in transfected cells has been confirmed. Three potential caveolin-1-binding sites were reported to be present in the 3a protein. By using various biochemical, biophysical and genetic techniques, interaction of the 3a protein with caveolin-1 is demonstrated. Any one of the potential sites in the 3a protein was sufficient for this interaction. These results are discussed with respect to the possible roles of the 3a protein in the viral life cycle.


title: Care for adult non-ICU Covid-19 patients: early experiences from a Belgian tertiary care centre.

text: The current Covid-19 outbreak poses many challenges on hospital organisation and patient care. Our hospital lies at the epicentre of the Belgian epidemic. On April 1st, a total of 235 Covid-19 patients had been admitted to our hospital. This demanded an unprecedented adaptation of our hospital organisation, and we have met many clinical issues in the care for Covid-19 patients. In this article, we share our experience in the handling of some of the practical and organisational issues in the care for Covid-19 patients.


title: Homo-oligomerization of the porcine reproductive and respiratory syndrome virus nucleocapsid protein and the role of disulfide linkages.

text: As a step toward understanding the assembly pathway of the porcine reproductive and respiratory syndrome virus (PRRSV), the oligomeric properties of the nucleocapsid (N) protein were investigated. In this study, we have demonstrated that under nonreducing conditions the N protein forms disulfide-linked homodimers. However, inclusion of an alkylating agent (N-ethylmaleimide [NEM]) prevented disulfide bond formation, suggesting that these intermolecular disulfide linkages were formed as a result of spurious oxidation during cell lysis. In contrast, N protein homodimers isolated from extracellular virions were shown to have formed NEM-resistant intermolecular disulfide linkages, the function of which is probably to impart stability to the virion. Pulse-chase analysis revealed that N protein homodimers become specifically disulfide linked within the virus-infected cell, albeit at the later stages of infection, conceivably when the virus particle buds into the oxidizing environment of the endoplasmic reticulum. Moreover, NEM-resistant disulfide linkages were shown to occur only during productive PRRSV infection, since expression of recombinant N protein did not result in the formation of NEM-resistant disulfide-linked homodimers. Mutational analysis indicated that of the three conserved cysteine residues in the N protein, only the cysteine at position 23 was involved in the formation of disulfide linkages. The N protein dimer was shown to be stable both in the presence and absence of intermolecular disulfide linkages, indicating that noncovalent interactions also play a role in dimerization. Non-disulfide-mediated N protein interactions were subsequently demonstrated both in vitro by the glutathione S-transferase (GST) pull-down assay and in vivo by the mammalian two-hybrid assay. Using a series of N protein deletion mutants fused to GST, amino acids 30 to 37 were shown to be essential for N-N interactions. Furthermore, since RNase A treatment markedly decreased N protein-binding affinity, it appears that at least in vitro, RNA may be involved in bridging N-N interactions. In cross-linking experiments, the N protein was shown to assemble into higher-order structures, including dimers, trimers, tetramers, and pentamers. Together, these findings demonstrate that the N protein possesses self-associative properties, and these likely provide the basis for PRRSV nucleocapsid assembly.


title: Wild animal surveillance for coronavirus HKU1 and potential variants of other coronaviruses.

text: 1. Although CoV-HKU1 was not identified in any of the studied animals, a coronavirus closely related to SARS-CoV (bat-SARS-CoV) was identified in 23 (19%) of 118 wild Chinese horseshoe bats by reverse transcriptase polymerase chain reaction (RT-PCR). 2. Complete genome sequencing and phylogenetic analysis showed that bat-SARS-CoV formed a distinct cluster with SARS-CoV as group 2b coronaviruses, distantly related to known group 2 coronaviruses. 3. Most differences between the bat-SARS-CoV and SARS-CoV genomes were observed in the spike gene. The presence of a29-bp insertion in ORF 8 of bat-SARS-CoV genome, not in most human SARS-CoV genomes, suggests that it has a common ancestor with civet SARS-CoV. 4. Antibody against recombinant bat-SARS-CoV nucleocapsid protein was detected in 84% of Chinese horseshoe bats using an enzyme immunoassay.Neutralising antibody to human SARS-CoV was also detected in those with lower viral loads.5. This study also revealed a previously unknown diversity of coronaviruses in bats, which are important natural reservoir for coronaviruses including SARS-CoV-like viruses.


title: [THE PSYCHOLOGICAL IMPACT OF A PANDEMIC OUTBREAK ON MEDICAL STAFF - LITERATURE REVIEW].

text: INTRODUCTION In January 2020, the World Health Organization (WHO) declared an international emergency due to the outbreak of COVID-19. In March 2020 the WHO announced that the epidemic could be characterized as a pandemic. Subsequently, the WHO acknowledged the importance of providing support to the medical teams, realizing the enormous burden and stress facing this group. The aim of the present study is to review the up-to-date professional literature dealing with the psychological impact of a pandemic outbreak on medical staff. The literature existing so-far is scarce. It does, however, provide evidence of the short- and long-term impact of such epidemics on the mental health of the medical teams. Direct exposure to affected patients, the risk of contagion, a subjectively negative perception of one's medical condition and being in isolation or quarantine, all constitute risk factors for mental symptoms such as depression, anxiety and post-trauma. A sense of control, especially regarding the means of protection and controlling infection, altruistic acceptance of the work-related risks, social support, good team-spirit in the medical unit and support from superiors are all protective factors.


title: Managing Prostate Cancer Surgical Patients during the COVID-19 Pandemic: A Brief Report of the Duke Cancer Institute's Initial Experience.

text: The coronavirus disease 2019 pandemic has rapidly placed tremendous stress on health systems around the world. In response, multiple health systems have postponed elective surgeries in order to conserve hospital beds and personal protective equipment, minimize patient traffic, and prevent unnecessary utilization and exposure of healthcare workers. The American College of Surgeons released the following statement on March 13, 2020: "Each hospital, health system and surgeon should thoughtfully review all scheduled elective procedures with a plan to minimize, postpone, or cancel electively scheduled operations, endoscopes, or other invasive procedures until we have passed the predicted inflection point in the exposure graph and can be confident that our health care infrastructure can support a potentially rapid and overwhelming uptick in critical patient care needs." In our state, North Carolina, Governor Roy Cooper requested that all hospitals postpone elective and non-urgent procedures and surgeries effective March 23, 2020.


title: Sequencing of coronavirus IBV genomic RNA: three open reading frames in the 5' 'unique' region of mRNA D.

text: The nucleotide sequence of a genomic cDNA clone corresponding to the 5' terminal domain of mRNA D of the Beaudette strain of infectious bronchitis virus (IBV) has been determined. This region contains three open reading frames which predict polypeptides of molecular weights 6700 (6.7K), 7.4K and 12.4K. The predicted 12.4K polypeptide has a codon usage very similar to that predicted for the products of the IBV nucleocapsid, membrane and spike genes. The sequence also predicts a hydrophobic, potentially membrane-anchoring, region in the N terminal half of the 12.4K polypeptide, and a hydrophilic C terminus.


Feline coronavirus in the intestinal contents of cats with feline infectious peritonitis.


title: Severe acute respiratory syndrome in Singapore.

text: AIMS To describe the epidemiological and clinical features of paediatric severe acute respiratory syndrome (SARS) in Singapore. METHODS The following data were retrospectively collected from the case records of all 71 patients (aged 7 months to 14 years) admitted from 23 March to 22 May 2003 to the SARS paediatric unit: patient demographics, contact history, clinical features, physiological parameters, investigations, treatment, and outcome. Using WHO criteria there were seven probable (P), 23 suspect (S), and 41 observe (O) cases. RESULTS Compared to the O cases P patients had a longer mean duration of fever (3.66 (SD 2.3) v 8.57 (SD 2.44) days), lower mean thrombocytopenia (248.3 (SD 82.7) v 173.7 (SD 49.0)x10(9)/l), leucopenia (8.19 (SD 4.45) v 3.06 (SD 1.02)x10(9)/l), lymphopenia (2.79 (SD 1.97) v 1.44 (SD 0.75)x10(9)/l), and neutropenia (4.48 (SD 2.88) v 1.24 (SD 0.43)x10(9)/l). Chest auscultation was abnormal in 71% of P patients, with mild crepitations detected. All had abnormal chest radiographs versus 39% of S cases, and 27% of O cases. CONCLUSIONS There are no distinguishing clinical features of paediatric SARS. The diagnosis is suggested by the paucity of clinical signs with an abnormal chest radiograph, and laboratory evidence of leucopenia, lymphopenia, and thrombocytopenia.