Paradoxical effects of cigarette smoke and COPD on SARS-CoV-2 infection and disease.

BACKGROUND: How cigarette smoke (CS) and chronic obstructive pulmonary disease (COPD) affect severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) infection and severity is controversial. We investigated the effects of COPD and CS on the expression of SARS-CoV-2 entry receptor ACE2 in vivo in COPD patients and controls and in CS-exposed mice, and the effects of CS on SARS-CoV-2 infection in human bronchial epithelial cells in vitro. METHODS: We quantified: (1) pulmonary ACE2 protein levels by immunostaining and ELISA, and both ACE2 and/or TMPRSS2 mRNA levels by RT-qPCR in two independent human cohorts; and (2) pulmonary ACE2 protein levels by immunostaining and ELISA in C57BL/6 WT mice exposed to air or CS for up to 6 months. The effects of CS exposure on SARS-CoV-2 infection were evaluated after in vitro infection of Calu-3 cells and differentiated human bronchial epithelial cells (HBECs), respectively. RESULTS: ACE2 protein and mRNA levels were decreased in peripheral airways from COPD patients versus controls but similar in central airways. Mice exposed to CS had decreased ACE2 protein levels in their bronchial and alveolar epithelia versus air-exposed mice. CS treatment decreased viral replication in Calu-3 cells, as determined by immunofluorescence staining for replicative double-stranded RNA (dsRNA) and western blot for viral N protein. Acute CS exposure decreased in vitro SARS-CoV-2 replication in HBECs, as determined by plaque assay and RT-qPCR. CONCLUSIONS: ACE2 levels were decreased in both bronchial and alveolar epithelial cells from COPD patients versus controls, and from CS-exposed versus air-exposed mice. CS-pre-exposure potently inhibited SARS-CoV-2 replication in vitro. These findings urge to investigate further the controversial effects of CS and COPD on SARS-CoV-2 infection.

Paradoxical effects of cigarette smoke and COPD on SARS-CoV2 infection and disease.

INTRODUCTION: How cigarette smoke (CS) and chronic obstructive pulmonary disease (COPD) affect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and severity is controversial. We investigated the protein and mRNA expression of SARS-CoV-2 entry receptor ACE2 and proteinase TMPRSS2 in lungs from COPD patients and controls, and lung tissue from mice exposed acutely and chronically to CS. Also, we investigated the effects of CS exposure on SARS-CoV-2 infection in human bronchial epithelial cells. METHODS: In Cohort 1, ACE2-positive cells were quantified by immunostaining in FFPE sections from both central and peripheral airways. In Cohort 2, we quantified pulmonary ACE2 protein levels by immunostaining and ELISA, and both ACE2 and TMPRSS2 mRNA levels by RT-qPCR. In C57BL/6 WT mice exposed to air or CS for up to 6 months, pulmonary ACE2 protein levels were quantified by triple immunofluorescence staining and ELISA. The effects of CS exposure on SARS-CoV-2 infection were evaluated after 72hr in vitro infection of Calu-3 cells. After SARS-CoV-2 infection, the cells were fixed for IF staining with dsRNA-specific J2 monoclonal Ab, and cell lysates were harvested for WB of viral nucleocapsid (N) protein. Supernatants (SN) and cytoplasmic lysates were obtained to measure ACE2 levels by ELISA. RESULTS: In both human cohorts, ACE2 protein and mRNA levels were decreased in peripheral airways from COPD patients versus both smoker and NS controls, but similar in central airways. TMPRSS2 levels were similar across groups. Mice exposed to CS had decreased ACE2 protein levels in their bronchial and alveolar epithelia versus air-exposed mice exposed to 3 and 6 months of CS. In Calu3 cells in vitro, CS-treatment abrogated infection to levels below the limit of detection. Similar results were seen with WB for viral N protein, showing peak viral protein synthesis at 72hr. CONCLUSIONS: ACE2 levels were decreased in both bronchial and alveolar epithelial cells from uninfected COPD patients versus controls, and from CS-exposed versus air-exposed mice. CS-pre-treatment did not affect ACE2 levels but potently inhibited SARS-CoV-2 replication in this in vitro model. These findings urge to further investigate the controversial effects of CS and COPD on SARS-CoV2 infection.

Mechanisms of root reinforcement in soils: an experimental methodology using four-dimensional X-ray computed tomography and digital volume correlation.

Vegetation on railway or highway slopes can improve slope stability through the generation of soil pore water suctions by plant transpiration and mechanical soil reinforcement by the roots. To incorporate the enhanced shearing resistance and stiffness of root-reinforced soils in stability calculations, it is necessary to understand and quantify its effectiveness. This requires integrated and sophisticated experimental and multi-scale modelling approaches to develop an understanding of the processes at different length scales, from individual root-soil interaction through to full soil-profile or slope scale. One of the challenges with multi-scale models is ensuring that they sufficiently closely represent real behaviour. This requires calibration against detailed high-quality and data-rich experiments. This study presents a novel experimental methodology, which combines in situ direct shear loading of a willow root-reinforced soil with X-ray computed tomography to capture the three-dimensional chronology of soil and root deformation within the shear zone. Digital volume correlation (DVC) analysis was applied to the computed tomography dataset to obtain full-field three-dimensional displacement and strain information. This paper demonstrates the feasibility and discusses the challenges associated with DVC experiments on root-reinforced soils.

Trace metals and persistent organic pollutants fingerprint on the particle flux in the deep subtropical NE Atlantic.

Particle flux material collected in 2000 m depth in the Northeast Atlantic at 33°N and 22°W was analyzed for trace metals and persistent organic pollutants. Element enrichment factors relative to lithogenic Al were elevated indicating possible anthropogenic contributions for all trace metals except V. Polycyclic aromatic hydrocarbons, polychlorinated biphenyls and the pesticide DDT exhibited median fluxes of 10.40 µg m-2d-1,0.29 µg m-2 d-1, and 0.90 µg m-2 d-1, respectively. Flux composition reflected long range transport, with low molecular weight and low-chlorinated compounds dominating ∑15PAH and ∑23PCB. PAH isomer ratios identified fossil fuel combustion as the main ∑15PAH source. The composition of ∑4DDT suggested inputs of the fresh technical pesticide during high dust intensity periods. Pollutant fluxes showed seasonality linked to export production in the region, as well as a dependence on annual and sub-annual dust input events.

Noise creates polarization artefacts.

The accuracy of calculations of both the degree and angle of polarization depend strongly on the noise in the measurements used. The noise in the measurements recorded by both camera based systems and spectrometers can lead to significant artefacts and incorrect conclusions about high degrees of polarization when in fact none exist. Three approaches are taken in this work: firstly, the absolute error introduced as a function of the signal to noise ratio for polarization measurements is quantified in detail. An important finding here is the reason for why several studies incorrectly suggest that black (low reflectivity) objects are highly polarized. The high degree of polarization is only an artefact of the noise in the calculation. Secondly, several simple steps to avoid such errors are suggested. Thirdly, if these points can not be followed, two methods are presented for mitigating the effects of noise: a maximum likelihood estimation method and a new denoising algorithm to best calculate the degree of polarization of natural polarization information.

On the high-pressure phase stability and elastic properties of ß-titanium alloys.

We have studied the compressibility and stability of different ß-titanium alloys at high pressure, including binary Ti-Mo, Ti-24Nb-4Zr-8Sn (Ti2448) and Ti-36Nb-2Ta-0.3O (gum metal). We observed stability of the ß phase in these alloys to 40 GPa, well into the ω phase region in the P-T diagram of pure titanium. Gum metal was pressurised above 70 GPa and forms a phase with a crystal structure similar to the η phase of pure Ti. The bulk moduli determined for the different alloys range from 97 ± 3 GPa (Ti2448) to 124 ± 6 GPa (Ti-16.8Mo-0.13O).

Crossover between liquidlike and gaslike behavior in CH_{4} at 400 K.

We report experimental evidence for a crossover between a liquidlike state and a gaslike state in fluid methane (CH_{4}). This crossover is observed in all of our experiments, up to a temperature of 397 K, 2.1 times the critical temperature of methane. The crossover has been characterized with both Raman spectroscopy and x-ray diffraction in a number of separate experiments, and confirmed to be reversible. We associate this crossover with the Frenkel line-a recently hypothesized crossover in dynamic properties of fluids extending to arbitrarily high pressure and temperature, dividing the phase diagram into separate regions where the fluid possesses liquidlike and gaslike properties. On the liquidlike side the Raman-active vibration increases in frequency linearly as pressure is increased, as expected due to the repulsive interaction between adjacent molecules. On the gaslike side this competes with the attractive van der Waals potential leading the vibration frequency to decrease as pressure is increased.

Low frequency sound propagation in activated carbon.

Activated carbon can adsorb and desorb gas molecules onto and off its surface. Research has examined whether this sorption affects low frequency sound waves, with pressures typical of audible sound, interacting with granular activated carbon. Impedance tube measurements were undertaken examining the resonant frequencies of Helmholtz resonators with different backing materials. It was found that the addition of activated carbon increased the compliance of the backing volume. The effect was observed up to the highest frequency measured (500 Hz), but was most significant at lower frequencies (at higher frequencies another phenomenon can explain the behavior). An apparatus was constructed to measure the effective porosity of the activated carbon as well as the number of moles adsorbed at sound pressures between 104 and 118 dB and low frequencies between 20 and 55 Hz. Whilst the results were consistent with adsorption affecting sound propagation, other phenomena cannot be ruled out. Measurements of sorption isotherms showed that additional energy losses can be caused by water vapor condensing onto and then evaporating from the surface of the material. However, the excess absorption measured for low frequency sound waves is primarily caused by decreases in surface reactance rather than changes in surface resistance.

The pressure-temperature phase diagram of MgH2 and isotopic substitution.

Computational thermodynamics using density functional theory ab initio codes is a powerful tool for calculating phase diagrams. The method is usually applied at the standard pressure of p = 1 bar and where the Gibbs energy is assumed to be equal to the Helmholtz energy. In this work, we have calculated the Gibbs energy in order to study the release temperature and phase modifications of MgH(2) at high pressures up to 10 GPa (100 kbar). The isotopic substitution of hydrogen with deuterium (or tritium) does not bring about any strong effects on the phase diagram. These considerations are of extreme importance for (i) the synthesis of novel substitutional magnesium based materials at high pressure and (ii) the determination of the correct reference states for the calculation of phase diagrams at high pressure. The calculated results are compared with experimental data obtained with an in situ neutron diffraction measurement.

Effect of blood product transfusion-induced tolerance on incidence of cardiac allograft rejection.

BACKGROUND: Blood product transfusion has been successfully used in solid-organ transplantation to induce tolerance. Whether a similar protective effect of blood product transfusion exists in heart transplantation is controversial. OBJECTIVE: To investigate the effect of cellular blood product transfusion within 2 weeks posttransplantation on the incidence of cellular and antibody-mediated rejection. PATIENTS AND METHODS: Patients were grouped on the basis of number of blood transfusions; group 1 received no transfusions, and groups 2, 3, and 4 each received an incremental number of transfusion units. All endomyocardial biopsy samples were routinely studied using immunofluorescence in the first 12 weeks posttransplantation. RESULTS: Baseline characteristics including age, sex, body mass index, history of diabetes, donor characteristics, and pretransplantation laboratory values were similar except that group 4 had a higher rate of previous sternotomy and longer ischemic time during transplantation. Approximately 9200 endomyocardial biopsy samples composed the data. Short- and long-term freedom from the International Society for Heart & Lung Transplantation grade 3A or higher cellular rejection and from antibody-mediated rejection were comparable between groups. CONCLUSIONS: Blood transfusions within the first 2 weeks post-transplantation do not seem to confer any protective effect against posttransplantation cellular rejection or antibody- mediated rejection. Whether other unmeasured confounding factors mask their effect requires further prospective studies.

Reproductive factors and specific histological types of breast cancer: prospective study and meta-analysis.

Little is known about how reproductive factors affect the risk of breast cancers of different histology. In an analysis of prospective data on 1.2 million middle-aged UK women, we used proportional hazards models to estimate the relative risks of six histological types in relation to menarche, childbearing and menopause. During 8.7 million person-years of follow-up, 17 923 ductal, 3332 lobular, 1062 tubular, 944 mixed ductal lobular, 330 mucinous and 117 medullary cancers were diagnosed. The effect of both age at menarche and age at first birth was greatest for lobular tumours; relative risks per 5-year increase in age at menarche for ductal, lobular, and tubular cancers were 0.93 (0.87-0.99), 0.65 (0.56-0.76), and 0.75 (0.57-0.98), respectively (P-value for heterogeneity=0.0001); and the relative risks per 5-year increase in age at first birth were 1.10 (1.07-1.12), 1.23 (1.17-1.29), and 1.13 (1.03-1.23), respectively (P-value for heterogeneity=0.0006). Increasing parity reduced the risk of each tumour type, except medullary cancers, but the reduction in risk was greater for mucinous cancers than for any other subtype considered (P<0.05 for comparison with each other subtype in turn). The effect of menopause did not vary significantly by tumour histology. Meta-analysis of published results on the effects of age at menarche and age at first birth on ductal and lobular cancers were in keeping with our findings.

Polymeric gene delivery of ischemia-inducible VEGF significantly attenuates infarct size and apoptosis following myocardial infarct.

The development of clinically beneficial myocardial gene therapy has been slowed by reliance on the use of viral carriers and non-physiologic, constitutive gene expression. To specifically address these issues, we have developed a non-viral gene carrier, water-soluble lipopolymer (WSLP), and an ischemia-inducible plasmid construct expressing vascular endothelial growth factor (VEGF), pRTP801-VEGF, to treat myocardial ischemia and infarction. Rabbits underwent ligation of the circumflex artery followed by injection of (a) an ischemia-inducible VEGF gene construct in a WSLP carrier; (b) a constitutively expressed, or unregulated, SV-VEGF gene construct in a WSLP carrier; (c) WSLP carrier alone; or (d) no injection therapy. Following 4 weeks treatment, ligation alone resulted in infarction of 48+/-7% of the left ventricle. With injection of WSLP carrier alone, 49+/-6% of the left ventricle was infarcted (P=NS). The constitutively expressed gene construct, SV-VEGF, reduced the infarct size to 32+/-7% of the left ventricle (P=0.007). The ischemia-inducible gene construct, RTP801-VEGF, further reduced the infarct size to 13+/-4% of the left ventricle (P<0.001). The use of a non-viral carrier to deliver an ischemia-inducible VEGF construct is effective in the treatment of acutely ischemic myocardium.

Hypoxia-inducible VEGF gene delivery to ischemic myocardium using water-soluble lipopolymer.

Therapeutic angiogenesis with gene encoding vascular endothelial growth factor (VEGF) is a new potential treatment in cardiovascular disease. However, unregulated VEGF-mediated angiogenesis has the potential to promote tumor growth, accelerate diabetic proliferative retinopathy, and promote rupture of atherosclerotic plaque. To be safe and effective, gene therapy with VEGF must be regulated. To limit the risk of pathological angiogenesis, we developed a hypoxia-inducible VEGF gene therapy system using the erythropoietin (Epo) enhancer and water-soluble lipopolymer (WSLP). pEpo-SV-VEGF or pSV-VEGF-Epo was constructed by insertion of the Epo enhancer upstream of the Simian Virus 40 (SV40) promoter or downstream of the poly(A) signal of pSV-VEGF. In vitro transfection showed that pEpo-SV-VEGF, not pSV-VEGF-Epo, induced the VEGF expression in hypoxic cells. In addition, the VEGF protein, which was produced from the Epo-SV-VEGF-transfected and hypoxia-incubated cells, was able to enhance the proliferation of the endothelial cells. Injection of the pEpo-SV-VEGF/WSLP complex showed that the expression of VEGF was induced in ischemic myocardium, compared to normal myo-cardium. Therefore, with the localized induction of VEGF and the low cytotoxicity of WSLP, the pEpo-SV-VEGF/WSLP system may be helpful to eventually treat ischemic heart disease.

Water-soluble lipopolymer as an efficient carrier for gene delivery to myocardium.

Water-soluble lipopolymer (WSLP), which consisted of polyethylenimine (PEI, 1800 Da) and cholesterol, was characterized as a gene carrier to smooth muscle cells and myocardium. Acid-base titration showed that WSLP had a proton-buffering effect. The size of WSLP/plasmid DNA (pDNA) complex was around 70 nm. WSLP/pDNA complex was transfected to A7R5 cells, a smooth muscle cell line. WSLP showed the highest transfection at a 40/1 N/P ratio. WSLP has higher transfection efficiency than PEI (1800 and 25 000 Da), SuperFect, and lipofectamine. In addition, WSLP has less cytotoxicity than PEI (25 000 Da), SuperFect, and lipofectamine. Since WSLP has cholesterol moiety, it may utilize cellular cholesterol uptake pathway, in which low-density lipoprotein (LDL) is involved. An inhibition study with free cholesterol or low-density lipoprotein (LDL) showed that transfection was inhibited by cholesterol or LDL, suggesting that WSLP/pDNA complex is transfected to the cells through the cholesterol uptake pathway. To evaluate the transfection efficiency to myocardium, WSLP/pDNA complex was injected into the rabbit myocardium. WSLP showed higher transfection than PEI and naked pDNA. WSLP expressed the transgene for more than 2 weeks. In conclusion, WSLP is an efficient carrier for local gene transfection to myocardium, and useful in in vivo gene therapy.

Predictors of outcome of mammography in the National Health Service Breast Screening Programme.

BACKGROUND: Little is known about the factors influencing the risk of recall for assessment, invasive diagnostic procedures, and early rescreening after screening mammography. METHODS: From June 1996 to March 1998 women attending screening at 10 National Health Service Breast Screening Programme (NHSBSP) centres completed a self administered questionnaire and were followed up for their screening outcome. RESULTS: 1969 (3.3%) out of 60 443 women aged 50-64 who had never used hormone replacement therapy (HRT) were recalled for assessment but were not diagnosed with breast cancer (defined here as false positive recall). After adjustment for the variation between centres, false positive recall was decreased significantly among women who were likely to have had a previous NHSBSP mammogram (odds ratio (OR) 0.49, 95% confidence interval (95% CI) 0.38 to 0.63 for likely versus unlikely), who were postmenopausal (OR 0.65, 95% CI 0.56 to 0.76 for postmenopausal v premenopausal) and increased significantly for women reporting previous breast surgery (OR 1.64, 95% CI 1.42 to 1.89). Although false positive recall decreased significantly with parity and increasing body mass index, these effects were not large and no significant variation was found with age, education, family history of breast cancer, oral contraceptive use, sterilisation, exercise, smoking, or alcohol consumption. Altogether 655 (1.1%) women had an invasive diagnostic procedure; no personal characteristics were predictive of this outcome, 286(0.5%) were referred for early rescreening, and this was increased significantly by nulliparity and a family history of breast cancer. INTERPRETATION: Premenopausal women, those without a previous NHSBSP mammogram, and women with previous breast surgery have an increased risk of false positive recall by the NHSBSP.

Risk factors for oesophageal, lung, oral and laryngeal cancers in black South Africans.

The authors used data collected from 1995 to 1999, from an on-going cancer case-control study in greater Johannesburg, to estimate the importance of tobacco and alcohol consumption and other suspected risk factors with respect to cancer of the oesophagus (267 men and 138 women), lung (105 men and 41 women), oral cavity (87 men and 37 women), and larynx (51 men). Cancers not associated with tobacco or alcohol consumption were used as controls (804 men and 1370 women). Tobacco smoking was found to be the major risk factor for all of these cancers with odds ratios ranging from 2.6 (95% CI 1.5-4.5) for oesophageal cancer in female ex-smokers to 50.9 (95% CI 12.6-204.6) for lung cancer in women, and 23.9 (95% CI 9.5-60.3) for lung cancer and 23.6 (95% CI 4.6-121.2) for laryngeal cancer in men who smoked 15 or more grams of tobacco a day. This is the first time an association between smoking and oral and laryngeal cancers has been shown in sub-Saharan Africa. Long-term residence in the Transkei region in the southeast of the country continues to be a risk factor for oesophageal cancer, especially in women (odds ratio=14.7, 95% CI 4.7-46.0), possibly due to nutritional factors. There was a slight increase in lung cancer (odds ratio=2.9, 95% CI 1.1-7.5) in men working in 'potentially noxious' industries. 'Frequent' alcohol consumption, on its own, caused a marginally elevated risk for oesophageal cancer (odds ratio=1.7, 95% CI 1.0-2.9, for women and odds ratio=1.8, 95% CI 1.2-2.8, for men). The risks for oesophageal cancer in relation to alcohol consumption increased significantly in male and female smokers (odds ratio=4.7, 95% CI=2.8-7.9 in males and odds ratio=4.8, 95% CI 3.2-6.1 in females). The above results are broadly in line with international findings.

Vitamin K and childhood cancer: analysis of individual patient data from six case-control studies.

To investigate the hypothesis that neonates who receive intramuscular vitamin K are at an increased risk of developing cancer, particularly leukaemia, a pooled analysis of individual patient data from six case-control studies conducted in Great Britain and Germany has been undertaken. Subjects comprised 2431 case children diagnosed with cancer before 15 years of age and 6338 control children. The retrospective assessment of whether or not an individual baby received vitamin K is not straightforward. In many cases no record was found in stored medical notes and two types of analysis were therefore conducted; in the first it was assumed that where no written record of vitamin K was found it had not been given, and in the second, where no written record of administration was found, information on hospital policy and perinatal morbidity was used to 'impute' whether or not vitamin K had been given. In the first analysis, no association was found between neonatal administration of intramuscular. vitamin K and childhood cancer: odds ratios adjusted for mode of delivery, admission to special care baby unit and low birth weight were 1.09 (95% confidence interval 0.92-1.28) for leukaemia and 1.05 (0.92-1.20) for other cancers. In the second analysis, the adjusted odds ratios increased to 1.21 (1.02-1.44) for leukaemia and 1.10 (0.95-1.26) for other cancers. This shift did not occur in all studies, and when data from the hypothesis generating Bristol study were excluded, the adjusted odds ratios for leukaemia became 1.06 (0.89-1.25) in the first analysis and 1.16 (0.97-1.39) when data on prophylaxis imputed from hospital policy and perinatal morbidity were used. We conclude that whilst the broad nature of the diagnostic groups and the poor quality of some of the vitamin K data mean that small effects cannot be entirely ruled out, our analysis provides no convincing evidence that intramuscular vitamin K is associated with childhood leukaemia.

Subjective assessment of left ventricular preload using transesophageal echocardiography: corresponding pulmonary artery occlusion pressures.

OBJECTIVE: To record pulmonary artery occlusion pressures (PAOPs) in patients whose left ventricular preload reserve was subjectively determined using transesophageal echocardiography (TEE). DESIGN: Prospective, blinded, nonrandomized. SETTING: University hospital. PARTICIPANTS: Twenty-three patients with well-preserved left ventricular function during nonemergent cardiac surgery. INTERVENTIONS: After separation from cardiopulmonary bypass, patients received repeated boluses of fluid volume through the aortic inflow cannula while being monitored with TEE. The endpoint for this fluid administration was a plateau in left ventricular fractional area change and end-diastolic area. This point at which additional fluid failed to cause noticeable increases in left ventricular end-diastolic area and fractional area change was defined as the preload reserve volume. After reaching the preload reserve volume, the PAOP was measured, as were the systolic blood pressure, left ventricular fractional area change, and end-diastolic area. MEASUREMENTS AND MAIN RESULTS: The mean PAOP for all patients at the time of achieving preload reserve volume was 18.6 +/- 2.9 mmHg. In 8 patients, the PAOP corresponding to preload reserve volume was elevated (20 to 25 mmHg). The remaining 15 patients had PAOPs ranging from 13 to 19 mmHg. When these 2 groups were compared with respect to left ventricular end-diastolic area, fractional area change, and systolic blood pressure, there were no significant differences between groups. The left ventricular wall thickness was significantly greater, however, in the group with elevated PAOP (1.37 +/- 0.04 cm) when compared with the group with normal ventricular filling pressures (1.05 +/- 0.15 cm) (p = 0.001). CONCLUSIONS: In patients with well-preserved left ventricular function and normal wall thickness, preload reserve volumes subjectively determined by TEE corresponded to a range of filling pressures historically targeted to maximize cardiac performance (13 to 19 mmHg). In a subset of patients with increased wall thickness, however, subjective determination of preload reserve was associated with filling pressures that were higher than traditionally considered optimal (20 to 25 mmHg). Similarities in left ventricular fractional area change and end-diastolic area between these 2 groups suggest that patients with elevated filling pressures had decreased ventricular compliance and were managed correctly with higher than usual PAOPs.