A Protocol Guide for the N. crassa Yeast Artificial Chromosome Library

A yeast artificial chromosome (YAC) library of Neurospora crassa strain 74-OR23-1A has been constructed. This library has been used to clone 750 kb of contiguous DNA sequences from the centromere region of linkage group VII (M. Centola and J. Carbon. 1994. Mol. Cell. Biol. 14:1510-1519). The purpose of this article is explicitly to outline procedures that have been developed for library screening and chromosome walking

Conservative Management of COVID-19 Patients—Emergency Palliative Care in Action

CONTEXT: The COVID-19 pandemic is spreading across the world. Many patients will not be suitable for mechanical ventilation owing to the underlying health conditions, and they will require a conservative approach including palliative care management for their important symptom burden. OBJECTIVES: To develop a management plan for patients who are not suitable for mechanical ventilation that is tailored to the stage their COVID-19 disease. METHODS: Patients were identified as being stable, unstable, or at the end of life using the early warning parameters for COVID-19. Furthermore, a COVID-19-specific assessment tool was developed locally, focusing on key symptoms observed in this population which assess dyspnoea, distress, and discomfort. This tool helped to guide the palliative care management as per patients' disease stage. RESULTS: A management plan for all patients' (stable, unstable, end of life) was created and implemented in acute hospitals. Medication guidelines were based on the limitations in resources and availability of drugs. Staff members who were unfamiliar with palliative care required simple, clear instructions to follow including medications for key symptoms such as dyspnoea, distress, fever, and discomfort. Nursing interventions and family involvement were adapted as per patients' disease stage and infection control requirements. CONCLUSION: Palliative care during the COVID-19 pandemic needs to adapt to an emergency style of palliative care as patients can deteriorate rapidly and require quick decisions and clear treatment plans. These need to be easily followed up by generalist staff members caring for these patients. Furthermore, palliative care should be at the forefront to help make the best decisions, give care to families, and offer spiritual support

One-Liners

One liners from: N.M. Martinez-Rossi, C. Andrade-Monteiro and S.R.C. Pombeiro; M. Orbach ; H. Liu and TJ. Schmidhauser; P.A. Hubbard and C.H. Wilso

Influence of water uptake on the aerosol particle light scattering coefficients of the Central European aerosol

The influence of aerosol water uptake on the aerosol particle light scattering was examined at the regional continental research site Melpitz, Germany. The scattering enhancement factor f(RH), defined as the aerosol particle scattering coefficient at a certain relative humidity (RH) divided by its dry value, was measured using a humidified nephelometer. The chemical composition and other microphysical properties were measured in parallel. f(RH) showed a strong variation, e.g. with values between 1.2 and 3.6 at RH=85% and λ=550 nm. The chemical composition was found to be the main factor determining the magnitude of f(RH), since the magnitude of f(RH) clearly correlated with the inorganic mass fraction measured by an aerosol mass spectrometer (AMS). Hysteresis within the recorded humidograms was observed and explained by long-range transported sea salt. A closure study using Mie theory showed the consistency of the measured parameters

Study of the relative humidity dependence of aerosol light-scattering in southern Spain

This investigation focuses on the characterisation of the aerosol particle hygroscopicity. Aerosol particle optical properties were measured at Granada, Spain, during winter and spring seasons in 2013. Measured optical properties included particle light-absorption coefficient (sap) and particle light-scattering coefficient (ssp) at dry conditions and at relative humidity (RH) of 85 +/- 10%. The scattering enhancement factor, f(RH=85%), had a mean value of 1.5 +/- 0.2 and 1.6 +/- 0.3 for winter and spring campaigns, respectively. Cases of high scattering enhancement were more frequent during the spring campaign with 27% of the f(RH=85%) values above 1.8, while during the winter campaign only 8% of the data were above 1.8. A Saharan dust event (SDE), which occurred during the spring campaign, was characterised by a predominance of large particles with low hygroscopicity. For the day when the SDE was more intense, a mean daily value of f(RH=85%)=1.3 +/- 0.2 was calculated. f(RH=85%) diurnal cycle showed two minima during the morning and afternoon traffic rush hours due to the increase in non-hygroscopic particles such as black carbon and road dust. This was confirmed by small values of the single-scattering albedo and the scattering Angstrom exponent. A significant correlation between f(RH=85%) and the fraction of particulate organic matter and sulphate was obtained. Finally, the impact of ambient RH in the aerosol radiative forcing was found to be very small due to the low ambient RH. For high RH values, the hygroscopic effect should be taken into account since the aerosol forcing efficiency changed from -13W/m2 at dry conditions to -17W/m2 at RH=85%.This work was supported by the Andalusia Regional Government through projects P10-RNM-6299 and P12-RNM-2409; by the Spanish Ministry of Economy and Competitiveness through projects CGL2010-18782, CSD2007-00067, CGL2011-13580-E/CLI and CGL2011-16124-E; and by EU through ACTRIS project (EU INFRA-2010-1.1.16-262254).G. Titos was funded by the program FPI of the Spanish Ministry of Economy and Competitiveness – Secretariat of Science, Innovation and Development under grant BES-2011-043721

Characterization of the Temperature-Sensitive Mutations un-7 and png-1 in Neurospora crassa

The model filamentous fungus Neurospora crassa has been studied for over fifty years and many temperature-sensitive mutants have been generated. While most of these have been mapped genetically, many remain anonymous. The mutation in the N. crassa temperature-sensitive lethal mutant un-7 was identified by a complementation based approach as being in the open reading frame designated NCU00651 on linkage group I. Other mutations in this gene have been identified that lead to a temperature-sensitive morphological phenotype called png-1. The mutations underlying un-7 result in a serine to phenylalanine change at position 273 and an isoleucine to valine change at position 390, while the mutation in png-1 was found to result in a serine to leucine change at position 279 although there were other conservative changes in this allele. The overall morphology of the strain carrying the un-7 mutation is compared to strains carrying the png-1 mutation and these mutations are evaluated in the context of other temperature-sensitive mutants in Neurospora

Exosomes Released from Mycoplasma Infected Tumor Cells Activate Inhibitory B Cells

Mycoplasmas cause numerous human diseases and are common opportunistic pathogens in cancer patients and immunocompromised individuals. Mycoplasma infection elicits various host immune responses. Here we demonstrate that mycoplasma-infected tumor cells release exosomes (myco+ exosomes) that specifically activate splenic B cells and induce splenocytes cytokine production. Induction of cytokines, including the proinflammatory IFN-γ and the anti-inflammatory IL-10, was largely dependent on the presence of B cells. B cells were the major IL-10 producers. In splenocytes from B cell deficient μMT mice, induction of IFN-γ+ T cells by myco+ exosomes was greatly increased compared with wild type splenocytes. In addition, anti-CD3-stimulated T cell proliferation was greatly inhibited in the presence of myco+ exosome-treated B cells. Also, anti-CD3-stimulated T cell signaling was impaired by myco+ exosome treatment. Proteomic analysis identified mycoplasma proteins in exosomes that potentially contribute to the effects. Our results demonstrate that mycoplasma-infected tumor cells release exosomes carrying mycoplasma components that preferentially activate B cells, which in turn, are able to inhibit T cell activity. These results suggest that mycoplasmas infecting tumor cells can exploit the exosome pathway to disseminate their own components and modulate the activity of immune cells, in particular, activate B cells with inhibitory activity