Hypoxia and reoxygenation do not upregulate adhesion molecules and natural killer cell adhesion on human endothelial cells in vitro

Objectives: Ischemia/reperfusion injury is characterized by endothelial cell activation leading to increased expression of adhesion molecules such as inter-cellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, endothelial- and platelet-selectin (E- and P-selectin), and to the subsequent recruitment of leukocytes. The aim of the present study was to investigate the respective effects of a proinflammatory cytokine (tumor necrosis factor alpha , TNF-α), hypoxia and/or reoxygenation on adhesion molecule expression and natural killer (NK) cell adhesion in an in vitro model of I/R. Methods: Human aortic endothelial cells (HAEC) were stimulated in vitro for 8h with TNF-α (1000 U/ml) and exposed to hypoxia (1% O2), reoxygenation (21% O2) or different combinations thereof. Cell surface expression of ICAM-1, VCAM-1 and E-/P-selectin on HAEC was analyzed by flow cytometry, and culture supernatants were tested for soluble adhesion molecules by ELISA. Rolling adhesion of NK cells on HAEC was determined using a rotating assay. Results: Untreated HAEC constitutively expressed ICAM-1 on their surface but neither expressed E-/P-selectin, VCAM-1, nor shedded soluble adhesion molecules. Exposure of HAEC to hypoxia or hypoxia and reoxygenation did not upregulate cell surface expression or shedding of adhesion molecules. In contrast, TNF-α significantly upregulated cell surface expression of ICAM-1, VCAM-1, and E-/P-selectin and led to the shedding of ICAM-1 and E-selectin. Combined treatment of HAEC with TNF-α, hypoxia and reoxygenation reduced E-/P-selectin surface expression and enhanced E-selectin shedding, but did not further influence ICAM-1 and VCAM-1. Soluble VCAM-1 was not detected. NK cell adhesion on HAEC increased 4-fold after TNF-α stimulation, but was not affected by hypoxia or hypoxia and reoxygenation. Conclusions: Both the expression of endothelial adhesion molecules and rolling NK cell adhesion was upregulated by TNF-α but not by hypoxia alone or hypoxia followed by reoxygenation supporting the view that anti-inflammatory treatment may reduce ischemia/reperfusion injur

Laparoscopic versus open splenectomy for nontraumatic diseases

BACKGROUND: Laparoscopic splenectomy (LS) is the standard procedure for normal size or moderately enlarged spleens; open splenectomy (OS) is preferred in cases of splenomegaly. In this study, indications for and complications of open and laparoscopic splenectomy were analyzed, with the aim to identify patients who will benefit from either technique. METHOD: A consecutive series of 52 patients undergoing elective open or laparoscopic splenectomy between January 2001 and December 2006 was analyzed. Spleen volume was calculated as length x width x depth from the pathologist's measurements. RESULTS: LS was performed in 25 patients with a median age of 41 years (range = 24-65). OS was performed in 27 patients with a median age of 60 years (range = 24-86) (p < 0.001). Conversion to OS was necessary in two patients (8%). Operation time was significantly shorter in LS (p < 0.05). Spleen volume was significantly greater in patients who underwent open (median = 2520 ml, range = 150-16,800 ml) versus laparoscopic (median = 648 ml, range = 150-4860 ml) splenectomy (p = 0.001). In 36% of all laparoscopic procedures, spleen volume exceeded 1000 ml. The underlying disease was mainly immunothrombocytopenia in LS patients and lymphoma and osteomyelofibrosis in OS patients. Five patients died after OS. Significantly more patients were hospitalized longer than 7 days following OS than following LS (p < 0.05). Overall complication rate was higher after OS (LS, 8; OS, 13 patients; p < 0.05). CONCLUSIONS: LS was preferred in younger patients with moderate splenomegaly, while massive splenomegaly mostly led to OS. In view of the absence of technique-related differences, LS can primarily be attempted in all patients

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Cells were either infected with 128 IFU/cell Cpn-K6 for 72 h (B,C) or incubated with staurosporin 1 μM for 16 h (D) or were left untreated (A)

<p><b>Copyright information:</b></p><p>Taken from "induces aponecrosis in human aortic smooth muscle cells"</p><p>BMC Microbiology 2005;5():2-2.</p><p>Published online 21 Jan 2005</p><p>PMCID:PMC547904.</p><p>Copyright © 2005 Dumrese et al; licensee BioMed Central Ltd.</p> A: Untreated HASMC with normal ultrastructure, note the elongated nucleus with finely dispersed euchromatin. B: HASMC with a typical Cpn inclusion. The membrane bound vesicle contains dark (= elementary bodies) and pale (= reticular bodies) stained bacteria. The chromatin structure in the nucleus is unchanged. C: Cpn infected HASMC undergoing cell death. The nucleus is characterized by highly condensed heterochromatin (arrow head) and a compact nucleolus and is surrounded by dilate organelles (arrow). The cell membrane is completely disrupted. D: Staurosporin treated HASMC. The nucleus is partially fragmented containing patchy condensed heterochromatin (arrow heads)

Cells infected with Cpn-K6 in doses between 8 – 386 IFU/cell or with Cpn-VR1310 in doses between 1 – 32 IFU/cell were stained with annexin-V / propidium iodide and analyzed by flow cytometry 48 h post infection

<p><b>Copyright information:</b></p><p>Taken from "induces aponecrosis in human aortic smooth muscle cells"</p><p>BMC Microbiology 2005;5():2-2.</p><p>Published online 21 Jan 2005</p><p>PMCID:PMC547904.</p><p>Copyright © 2005 Dumrese et al; licensee BioMed Central Ltd.</p> Mock inoculated cells are shown as controls. In figure A and B pooled data out of three independent experiments are displayed. A: The amount of single annexin-V positive cells increased dependent on the chlamydial infectious dose (-◆ -: Cpn-K6; -●-: Cpn-VR1310) but not in mock treated (-○-) cells. (*: significant compared to mock treated cells with p < 0.001) B: the number of single annexin-V positive cells increased over the time when cells were infected with Cpn-K6 128 IFU/cell (-●-) but not if they were mock treated (-○-) (significant compared to mock treated cells with p < 0.001)(#)). C: Dot plots of one representative experiment are depicted to illustrate the amount of annexin V / propidium iodide double labeled cells. Upper left panel: HASMC treated with staurosporin (apoptosis). Upper right panel: HASMC treated with Na-azide (necrosis). Middle left panel: HASMC infected with Cpn-VR1310 at 32 IFU / cell 48 h post infection. Middle right panel: HASMC infected with Cpn-K6 at 128 IFU / cell 48 h post infection. Lower left panel: mock treated HASMC

HASMC were infected with Cpn-K6 128 IFU/cell (A, B, C, D), Cpn-VR1310 32 IFU/cell (E, F) for 72 h or were inoculated with mock lysate (G) or treated with staurosporin 1 μM for 14 h (H)

<p><b>Copyright information:</b></p><p>Taken from "induces aponecrosis in human aortic smooth muscle cells"</p><p>BMC Microbiology 2005;5():2-2.</p><p>Published online 21 Jan 2005</p><p>PMCID:PMC547904.</p><p>Copyright © 2005 Dumrese et al; licensee BioMed Central Ltd.</p> The cells were subsequently stained for TUNEL (green), NHS-biotin (brown), anti-Cpn-MOMP (red) and DNA (blue). A: HASMC bearing a Cpn-K6 inclusion. DAPI stained chromatin structure is normal and TUNEL as well as NHS-biotin labeling are negative. B: Cpn-K6 infected HASMC with multiple chlamydial spots (arrow heads) and normal nuclear structure. C, E: Cpn-K6 (C) or Cpn-VR1310 (E) infected HASMC with spot like infection (arrow heads). The condensed nucleus is TUNEL positive, the cytoplasm is diffusely labeled with NHS-biotin. D, F: HASMC with Cpn-K6 (D) or Cpn-VR1310 (F) in aggregated (arrow) spots and single spots (arrow heads). Note positive TUNEL labeling of the shrunken nucleus and NHS-biotin staining of the cytoplasm. G: Mock inoculated HASMC. Note the fine granular chromatin structure and lack of TUNEL- and cytoplasmic NHS-biotin staining. H: HASMC incubated with staurosporin. The nucleus exhibits a distinct nuclear fragmentation (DAPI staining) but no TUNEL labeling. The cytoplasm is NHS-biotin negative indicating an intact cell membrane. I: Quantification of TUNEL positive nuclei depending on the chlamydial infectious dose 72 h post infection reveals a dose dependent increase of TUNEL positive cells. Cells in 5 random fields with an area of 62'500 μmwere counted and the percentage of TUNEL positive cells was calculated. -▲-: Cpn-K6; -■-: Cpn-VR131

Delayed colorectal cancer care during covid-19 pandemic (decor-19). Global perspective from an international survey

Background The widespread nature of coronavirus disease 2019 (COVID-19) has been unprecedented. We sought to analyze its global impact with a survey on colorectal cancer (CRC) care during the pandemic. Methods The impact of COVID-19 on preoperative assessment, elective surgery, and postoperative management of CRC patients was explored by a 35-item survey, which was distributed worldwide to members of surgical societies with an interest in CRC care. Respondents were divided into two comparator groups: 1) ‘delay’ group: CRC care affected by the pandemic; 2) ‘no delay’ group: unaltered CRC practice. Results A total of 1,051 respondents from 84 countries completed the survey. No substantial differences in demographics were found between the ‘delay’ (745, 70.9%) and ‘no delay’ (306, 29.1%) groups. Suspension of multidisciplinary team meetings, staff members quarantined or relocated to COVID-19 units, units fully dedicated to COVID-19 care, personal protective equipment not readily available were factors significantly associated to delays in endoscopy, radiology, surgery, histopathology and prolonged chemoradiation therapy-to-surgery intervals. In the ‘delay’ group, 48.9% of respondents reported a change in the initial surgical plan and 26.3% reported a shift from elective to urgent operations. Recovery of CRC care was associated with the status of the outbreak. Practicing in COVID-free units, no change in operative slots and staff members not relocated to COVID-19 units were statistically associated with unaltered CRC care in the ‘no delay’ group, while the geographical distribution was not. Conclusions Global changes in diagnostic and therapeutic CRC practices were evident. Changes were associated with differences in health-care delivery systems, hospital’s preparedness, resources availability, and local COVID-19 prevalence rather than geographical factors. Strategic planning is required to optimize CRC care