Finchbalslev5388
The lactic acid bacterium Enterococcus faecalis genomic DNA and seven phylogenetically distant bacterial genomic DNAs were microinjected into 126 enlarged protoplasts of E. faecalis. After the microinjection, a time-lapse observation was performed on how the cells enlarged. Most cells did not stop enlarging. The enlargement patterns were compared with the enlargement of E. faecalis protoplasts not treated by microinjection (control). They were clustered into three groups, with different levels and speeds of protoplast enlargement. The statistical analyses showed that the protoplasts injected by E. faecalis and four of the seven phylogenetically different bacterial genomic DNAs had enlargement patterns significantly different from those of the control. Thus, injected genomic DNAs affected the protoplast enlargement. Most of the affected cells, including the E. faecalis genome, had weakened enlargement.We performed an in-depth analysis of the virucidal effect of discrete wavelengths UV-C (278 nm), UV-B (308 nm), UV-A (366 nm) and violet (405 nm) on SARS-CoV-2. By using a highly infectious titer of SARS-CoV-2 we observed that the violet light-dose resulting in a 2-log viral inactivation is only 104 times less efficient than UV-C light. Moreover, by qPCR (quantitative Polymerase chain reaction) and fluorescence in situ hybridization (FISH) approach we verified that the viral titer typically found in the sputum of COVID-19 patients can be completely inactivated by the long UV-wavelengths corresponding to UV-A and UV-B solar irradiation. The comparison of the UV action spectrum on SARS-CoV-2 to previous results obtained on other pathogens suggests that RNA viruses might be particularly sensitive to long UV wavelengths. Our data extend previous results showing that SARS-CoV-2 is highly susceptible to UV light and offer an explanation to the reduced incidence of SARS-CoV-2 infection seen in the summer season.
The link between inflammation and depression has been investigated extensively. Cognitive dysfunction in depression is an unmet treatment need. A better understanding of possible links between inflammation and cognition in people with depression may help to identify new treatment targets.
We report findings from a study comparing a range of cognitive functions between 80 depressed patients with (C-reactive protein ≥3mg/L; n=37) and without (CRP<3mg/L; n=43) evidence of inflammation. All participants met the International Classification of Diseases 10th Revision criteria for current depressive episode and had somatic symptoms of depression. All participants completed cognitive testing and clinical assessment and were screened for acute infection.
Patients with evidence of inflammation, compared to those without, had slower psychomotor speed as measured by symbol coding task (mean difference=0.06, 95% CI=0.003-0.11) and slower reaction time, as measured by a simple movement time task (mean difference=26.56, 95% CI=-48.92 to -4.20). These effects were fully explained after controlling for age, sex, and body mass index. Measures of emotional processing, memory, and planning were comparable between two groups.
Certain cognitive domains, particularly processing speed and reaction time may be more affected in depressed patients with evidence of low-grade inflammation and somatic symptoms. Further studies with larger samples are required for a clearer understanding of the association between inflammation and cognitive dysfunction in depression.
Certain cognitive domains, particularly processing speed and reaction time may be more affected in depressed patients with evidence of low-grade inflammation and somatic symptoms. Further studies with larger samples are required for a clearer understanding of the association between inflammation and cognitive dysfunction in depression.
Varied cutaneous manifestations of COVID-19 have been described, but most studies are based on photographic or application-based observations, without a direct observed-based evaluation by dermatologists.
To study the types of cutaneous manifestations of COVID-19 among confirmed inpatients admitted to COVID-19 wards and intensive care units (ICUs).
This cross-sectional analysis was conducted at a referral hospital in Delhi, India. Four hundred forty consecutive reverse transcription-polymerase chain reaction (RT-PCR)-confirmed cases diagnosed with moderate or severe COVID-19 and admitted to COVID-19 wards or ICUs, respectively, were included. A cutaneous finding was considered to be associated with COVID-19 if it had been described earlier as a consequence of COVID-19 and was observed at the time of or within the first 48hours of admission (after excluding drugs and comorbidities as causes).
Two hundred seventy patients with moderate COVID-19 were admitted to COVID-19 wards, whereas 170 with severe disease were admitted to ICUs.Only 7 of the 270 ward patients (2.59%) and 3 of the 170 ICU patients (1.76%) had cutaneous findings associated with COVID-19.
Cutaneous findings attributable to COVID-19 are infrequent, and we believe that these might have been overestimated or overemphasized in earlier studies. Although coagulopathic findings may be associated with severe COVID-19, causation cannot be established in this cross-sectional study.
Cutaneous findings attributable to COVID-19 are infrequent, and we believe that these might have been overestimated or overemphasized in earlier studies. Although coagulopathic findings may be associated with severe COVID-19, causation cannot be established in this cross-sectional study.Upon viral infection, several proteins in the innate signaling pathway form aggregates, which in turn promote the activation of innate antiviral immune response. In this protocol, we use herpes simplex virus type 1 (HSV-1) to infect mouse peritoneal macrophages, and show how to detect the aggregation of TBK1 upon viral infection. The protocol is adaptable for other proteins and other viruses. For complete details on the use and execution of this profile, please refer to Yan et al. (2021).Patient-derived tumor organoids can be predictive of patient's treatment responses, and normal tissue-derived organoids allow for drug toxicity testing. Combining both types of organoids therefore enables screening for tumor-specific drug vulnerabilities. Here, we provide a detailed protocol for organoid drug screening using, as proof-of-principle, patient-derived malignant rhabdoid tumor organoids. Marimastat solubility dmso The protocol can be adapted for drug testing on any tumor and/or normal tissue-derived organoid culture. For complete details on the use and execution of this protocol, please refer to Calandrini et al. (2021).Large, publicly available neuroimaging datasets are becoming increasingly common, but their use presents challenges because of insufficient knowledge of the tool options for data processing and proper data organization. Here, we describe a protocol to lessen these barriers. We describe the steps for the search and download of the open-source dataset. We detail the steps for proper data management and practical guidelines for data analysis. Finally, we give instructions for data and result sharing on public repositories and preprint services. For complete details on the use and execution of this profile, please refer to Horien et al. (2021).Mammalian cyclic dinucleotide 2'3'-cGAMP functions as a second messenger in innate immune response. Here, we report a protocol to utilize 2'3'-cGAMP photoaffinity probes to capture 2'3'-cGAMP-binding or 2'3'-cGAMP-interacting proteins from HeLa cell lysate for in-gel visualization by fluorescent imaging or identification by SILAC-based quantitative MS. Further validation is also executed using photoaffinity probes to demonstrate the direct interaction of 2'3'-cGAMP with purified target proteins in vitro or endogenous target proteins in 293T cells. For complete details on the use and execution of this profile, please refer to Hou et al. (2021).Proximity-dependent biotinylation (BioID) screens are excellent tools to capture in cellulo interactomes for a large variety of baits, including transient and weak affinity interactions, as well as localization-specific proximity components, which are much harder to detect with conventional approaches. Here, we describe the major starting steps and a detailed protocol on how to perform BioID in mammalian cells. We also describe the mass spectrometry procedure and the bioinformatics pipeline for the data analysis. For complete details on the use and execution of this profile, please refer to Bagci et al. (2020).Ubiquitin-fold modifier 1 (UFM1) system is a recently identified ubiquitin-like modification with essential biological functions. Similar to ubiquitination, the covalent conjugation of UFM1 (UFMylation) to target proteins involves a three-step enzymatic cascade catalyzed sequentially by UFM1-activating enzyme 5 (UBA5, E1), UFM1-conjugating enzyme 1 (UFC1, E2), and UFM1-specific ligase 1 (UFL1, E3). Here, we provide an optimized protocol adapted to previously reported methods for detecting the UFMylation of target protein in human cells and in vitro assays, respectively, with high reliability and reproducibility. For complete details on the use and execution of this protocol, please refer to Liu et al. (2020).Epithelial tissues sustain barrier function by removing and replacing aberrant or unfit cells. Here, we describe approaches to evaluate epithelial restorative capacity after inducing cell loss in zebrafish larvae. We provide details to quantify morphological changes to the tail fin epithelium after cell loss, and instructions to interrogate changes in gene expression and proliferation associated with replacement of the lost cells. Together, this approach establishes an in vivo vertebrate model for the rapid assessment of molecular pathways controlling epithelial regeneration. For complete details on the use and execution of this profile, please refer to Wurster et al. (2021).Fluorescent protein (FP)-based kinase activity biosensors are powerful tools for probing the spatiotemporal dynamics of signaling pathways in living cells. Yet, the limited sensitivity of most kinase biosensors restricts their reliable application in high-throughput detection modalities. Here, we report a protocol for using an ultrasensitive excitation-ratiometric PKA activity reporter, ExRai-AKAR2, to detect live-cell PKA activity via fluorescence microplate reading and epifluorescence microscopy. The high sensitivity of ExRai-AKAR2 is well suited to these high-throughput applications. For complete details on the use and execution of this protocol, please refer to Mehta et al. (2018) andZhang et al., 2021a) .We describe a protocol to conduct a high-throughput in vitro processing assay, using 1,881 human primary microRNAs (pri-miRNAs) and recombinant Microprocessor complex, followed by deep sequencing library generation. This comprehensive approach allows the mapping of cleavage sites and the measurement of processing efficiency of a large number of substrates simultaneously. Our protocol is readily modifiable to investigate the effects of chemicals and regulatory proteins. Moreover, cis-acting elements can be examined by replacing the wild-type pri-miRNAs with mutant variants. For complete details on the use and execution of this profile, please refer to Kim et al. (2021).
