Integration of MALDI FTICR MSI data with the open-source software Cytomine

editorial reviewedOur skin is constantly exposed to solar radiation, high oxygen levels, and environmental pollutants. Our study aims to target specific native (LipS) and oxidized phospholipids (oxLipS) that are known to be senescence-associated secretory phenotype (SASP) related (i.e., LysoPC, oxPAPC). The production of these oxidized species is obtained after exposure to UV light. Here, we employ MALDI FTICR mass spectrometry imaging (MSI) to visualize and identify lipid species of interest in an organotypic model system and to integrate this data with the open-source software Cytomine.COMULIS COST ACTIO

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

Srovnání sací bublinky a bandáže pro analýzu epidermálních genů, bílkovin a lipidů

Analysis of epidermal genes, proteins and lipids is important in the research and diagnosis of skin diseases. Although punch biopsy is the first-choice technique for the skin sampling, it is unnecessarily invasive for obtaining a sample just for the epidermal analysis. Here we compare two less invasive methods, suction blistering (SB) and tape stripping (TS), for the analysis of selected epidermal genes (quantitative real-time reverse transcription PCR, qRT-PCR), proteins (western blotting, WB), and lipids in ten healthy volunteers. TS provided significantly less material than SB and no viable epidermal layers could be obtained according to the reflectance confocal microscopy. Consistently, only the SC protein filaggrin and housekeeping GAPDH together with FLG and RPL13A mRNA were detected by TS. In the SB samples, WB and qRT-PCR could easily detect all the selected proteins (claudin-1, occludin, filaggrin, laminin and GAPDH) and genes (CLDN1, OCLN, FLG, LAMA3 and RPL13A), respectively. A single SB sample further provided enough of material for immunohistochemistry and lipid analyses, which was not feasible with the TS samples. Immunohistochemistry of the SB samples showed intact epidermal structure and a characteristic expression of claudin-1. Infrared spectroscopy showed well-ordered lipids with both orthorhombic and hexagonal packing and high-performance thin layer chromatography confirmed all lipid classes (including ceramide subclasses) in correct proportions. Taken together, SB represents a reliable sampling technique that can be utilized for multipurpose epidermal analyses in various studies.Analýza epidermálních genů, proteinů a lipidů je důležitá při výzkumu a diagnostice kožních onemocnění. Ačkoli biopsie punch je metoda první volby pro vzorkování kůže, je zbytečně invazivní pro získání vzorku jen pro epidermální analýzu. Pro analýzu vybraných epidermálních genů (kvantitativní PCR reverzní transkripce v reálném čase, qRT-PCR), bílkoviny (Western blotting, WB) a srovnávací metody (SB) a strip stripping (TS) lipidů u deseti zdravých dobrovolníků. TS poskytovaly výrazně méně než SB a žádné životaschopné epidermální vrstvy nemohou být získány podle reflexní konfokální mikroskopie. Konzistentně byly TS detekovány pouze SC proteiny filaggrinu a housekeeping GAPDH společně s FLG a RPL13A mRNA. U vzorků SB by mohly WB a qRT-PCR snadno detekovat všechny vybrané proteiny (claudin-1, okluzin, filaggrin, laminin a GAPDH) a geny (CLDN1, OCLN, FLG, LAMA3 a RPL13A). Jeden vzorek SB dále poskytl dostatek materiálu pro imunohistochemické a lipidové analýzy, což nebylo možné u vzorků TS. Imunohistochemie vzorků SB ukázala intaktní epidermální strukturu a charakteristickou expresi claudinu-1. Infračervená spektroskopie ukázala dobře uspořádané lipidy s orthorhombickým a hexagonálním obalem a vysoce výkonná tenkovrstvá chromatografie potvrdila všechny třídy lipidů (včetně ceramidových podtřížek) ve správném poměru. Společně SB představuje spolehlivou metodu vzorkování, která může být použita pro víceúčelové epidermální analýzy v různých studiích

Biophysical study of CdTe quantum dots interactions with albumin and antibody as the base of photodynamic therapy.

Photodynamic therapy is a relatively new type of cancer treatment, or eventually it is used at the elimination of undesirable pathogenic microorganisms. Quantum dots (QDs) are semiconducting nanocrystals with a size of 2-20 nm and can be used in photodynamic therapy. Two types of CdTe QDs were prepared by microwave synthesis (500 W), typical absorption spectra had maxima 'green = 554 nm, - yellow = 580 nm. The determined size of the generated nanoparticles ranged between 5 - 10 nm. CdTe QDs were further studied by fluorescence analysis at excitation wavelength - = 250 nm to obtain emission maxima (?em552) for green and (?em582) for yellow nanoparticles. Interaction study of CdTe QDs with bovine serum albumin (BSA) and with polyclonal chicken antibodies against sarcosine (AntiSar) was performed. BSA (100, 50, 25, 12.5, 6.3, 3.1, 1.6 and 0 'M) and AntiSar (20, 10, 5, 2.5, 1.3, 0.7, 0.3, and 0 g/L) were monitored with 1:1 addition of 50 'M QDsgreen or QDsyellow. The decrease in intensity of the normalized fluorescence signal in the presence of BSA or AntiSar was observed by 70-90%. The observed dependencies showed a linear trend (R2 = 0.9) with relative error of 9-12% calculated from 5 independent repetitions. In addition, it was possible to monitor the signal shift to shorter wavelengths at the highest applied BSA concentration by 4-16 nm. The obtained data suggest that the size of individual QDs will affect intensity of an interaction with biomolecule. Nanoconstructs should therefore be targeted according to these experimental data for their intended use. In further experiments, CdTe QDs modified with AntiSar will be used for targeted prostate cancer therapy using photodynamic effect