18S rRNA is a reliable normalisation gene for real time PCR based on influenza virus infected cells

Background: One requisite of quantitative reverse transcription PCR (qRT-PCR) is to normalise the data with an internal reference gene that is invariant regardless of treatment, such as virus infection. Several studies have found variability in the expression of commonly used housekeeping genes, such as beta-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), under different experimental settings. However, ACTB and GAPDH remain widely used in the studies of host gene response to virus infections, including influenza viruses. To date no detailed study has been described that compares the suitability of commonly used housekeeping genes in influenza virus infections. The present study evaluated several commonly used housekeeping genes [ACTB, GAPDH, 18S ribosomal RNA (18S rRNA), ATP synthase, H+ transporting, mitochondrial F1 complex, beta polypeptide (ATP5B) and ATP synthase, H+ transporting, mitochondrial Fo complex, subunit C1 (subunit 9) (ATP5G1)] to identify the most stably expressed gene in human, pig, chicken and duck cells infected with a range of influenza A virus subtypes. Results: The relative expression stability of commonly used housekeeping genes were determined in primary human bronchial epithelial cells (HBECs), pig tracheal epithelial cells (PTECs), and chicken and duck primary lung-derived cells infected with five influenza A virus subtypes. Analysis of qRT-PCR data from virus and mock infected cells using NormFinder and BestKeeper software programmes found that 18S rRNA was the most stable gene in HBECs, PTECs and avian lung cells. Conclusions: Based on the presented data from cell culture models (HBECs, PTECs, chicken and duck lung cells) infected with a range of influenza viruses, we found that 18S rRNA is the most stable reference gene for normalising qRT-PCR data. Expression levels of the other housekeeping genes evaluated in this study (including ACTB and GPADH) were highly affected by influenza virus infection and hence are not reliable as reference genes for RNA normalisation

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

Temporal feedback control of high-intensity laser pulses to optimize ultrafast heating of atomic clusters

We describe how active feedback routines can be applied at limited repetition rate (5 Hz) to optimize high- power ( P> 10 TW) laser interactions with clustered gases. Optimization of x-ray production from an argon cluster jet, using a genetic algorithm, approximately doubled the measured energy through temporal modification of the 150 mJ driving laser pulse. This approach achieved an increased radiation yield through exploration of a multi-dimensional parameter space, without requiring detailed apriori knowledge of the complex cluster dynamics. The optimized laser pulses exhibited a slow rising edge to the intensity profile, which enhanced the laser energy coupling into the cluster medium, compared to the optimally compressed FWHM pulse (40 fs). Our work suggests that this technique can be more widely utilized for control of intense pulsed secondary radiation from petawatt-class laser systems

Heating and lighting: understanding overlooked energy-consumption activities in the Indian residential sector

Understanding the climate impact of residential emissions starts with determining the fuel consumption of various household activities. While cooking emissions have been widely studied, non-cooking energy-consumption activities in the residential sector such as heating and lighting, have been overlooked owing to the unavailability of data at national levels. The present study uses data from the Carbonaceous Aerosol Emissions, Source Apportionment and Climate Impacts (COALESCE) project, which consists of residential surveys over 6000 households across 49 districts of India, to understand the energy consumed by non-cooking residential activities. Regression models are developed to estimate information in non-surveyed districts using demographic, housing, and meteorological data as predictors. Energy demand is further quantified and distributed nationally at a 4 × 4 km resolution. Results show that the annual energy consumption from non-cooking activities is 1106 [201] PJ, which is equal to one-fourth of the cooking energy demand. Freely available biomass is widely used to heat water on traditional stoves, even in the warmer regions of western and southern India across all seasons. Space heating (51%) and water heating (42%) dominate non-cooking energy consumption. In comparison, nighttime heating for security personnel (5%), partly-residential personal heating by guards, dominant in urban centers and kerosene lighting (2%) utilize minimal energy. Biomass fuels account for over 90% of the non-cooking consumption, while charcoal and kerosene make up the rest. Half of the energy consumption occurs during winter months (DJF), while 10% of the consumption occurs during monsoon, when kerosene lighting is the highest. Firewood is the most heavily used fuel source in western India, charcoal in the northern hilly regions, agricultural residues and dung cake in the Indo-Gangetic plains, and kerosene in eastern India. The study shows that ∼20% of residential energy consumption is on account of biomass-based heating and kerosene lighting activities