Unstained viable cell recognition in phase-contrast microscopy

AbstractIndividual cell recognition is a relevant task to be accomplished when single-ion microbeam irradiations are performed. At INFN-LNL facility cell visualization system is based on a phase-contrast optical microscope, without the use of any cell dye. Unstained cells are seeded in the special designed Petri dish, between two mylar foils, and at present the cell recognition is achieved manually by an expert operator. Nevertheless, this procedure is time consuming and sometimes it could be not practical if the amount of living cells to be irradiated is large. To reduce the time needed to recognize unstained cells on the Petri dish, it has been designed and implemented an automated, parallel algorithm. Overlapping ROIs sliding in steps over the captured grayscale image are firstly pre-classified and potential cell markers for the segmentation are obtained. Segmented objects are additionally classified to categorize cell bodies from other structures considered as sample dirt or background. As a result, cell coordinates are passed to the dedicated CELLView program that controls all the LNL single-ion microbeam irradiation protocol, including the positioning of individual cells in front of the ion beam. Unstained cell recognition system was successfully tested in experimental conditions with two different mylar surfaces. The recognition time and accuracy was acceptable, however, improvements in speed would be useful

Do riparian forest fragments provide ecosystem services or disservices in surrounding oil palm plantations?

Agricultural expansion across tropical regions is causing declines in biodiversity and altering ecological processes. However, in some tropical agricultural systems, conserving natural habitat can simultaneously protect threatened species and support important ecosystem services. Oil palm cultivation is expanding rapidly throughout the tropics but the extent to which non-crop habitat supports biodiversity and ecosystem services in these landscapes is poorly documented. We investigated whether riparian forest fragments (riparian reserves) provide a pest control service or increase pest activity (disservice) within oil palm dominated landscapes in Sabah, Malaysian Borneo. We assessed the activity of potential predators of pest herbivores using plasticine caterpillar mimics and quantified herbivory rates on oil palm fronds in areas with and without riparian reserves. We also manipulated the shape and colour of the mimics to assess the extent to which artificial pest mimics reflect a predatory response. The presence of riparian reserves increased the attack rate on mimics by arthropods, but not by birds. Our methodological study suggested attacks on artificial pest mimics provide a better indication of predatory activity for birds than for arthropod predators. Herbivory rates were also not significantly affected by the presence of a riparian reserve, but we found some evidence that herbivory rates may decrease as the size of riparian reserves increases. Overall, we conclude that riparian forest fragments of 30 – 50 m width on each side of the river are unlikely to provide a pest control service. Nevertheless, our results provide evidence that these riparian buffer strips do not increase the density of defoliating pests, which should reassure managers concerned about possible negative consequences of preserving riparian buffers

High brightness inductively coupled plasma source for high current focused ion beam applications

A high brightnessplasmaion source has been developed to address focused ion beam(FIB) applications not satisfied by the liquid metal ion source (LMIS) based FIB. The plasmaFIB described here is capable of satisfying applications requiring high mill rates (>100μm³/s) with non-gallium ions and has demonstrated imaging capabilities with sub- 100-nm resolution. The virtual source size, angular intensity, mass spectra, and energy spread of the source have been determined with argon and xenon. This magnetically enhanced, inductively coupled plasmasource has exhibited a reduced brightness(βr) of 5.4×10³Am⁻²sr⁻¹V⁻¹, with a full width half maximum axial energy spread (ΔE) of 10eV when operated with argon. With xenon, βr=9.1×10³Am⁻²sr⁻¹V⁻¹ and ΔE=7eV. With these source parameters, an optical column with sufficient demagnification is capable of forming a sub-25-nm spot size at 30keV and 1pA. The angular intensity of this source is nominally three orders of magnitude greater than a LMIS making the source more amenable to creating high current focused beams, in the regime where spherical aberration dominates the LMIS-FIB. The source has been operated on a two lens ion column and has demonstrated a current density that exceeds that of the LMIS-FIB for current greater than 50nA. Source lifetime and current stability are excellent with inert and reactive gases. Additionally, it should be possible to improve both the brightness and energy spread of this source, such that the (βr/ΔE₂) figure-of-merit could be within an order of magnitude of a LMIS

63 Napromienianie przedoperacyjne u chorych na raka dolnej części odbytnicy

Od marca 1996 do czerwca 1998, u 47 chorych na raka dolnej części odbytnicy (dolna granica guza do 6 cm od brzegu odbytu) zastosowano napromienianie przedoperacyjne dawką 25 Gy we frakcjach po 5 Gy podaną w czasie tygodnia. Operację wykonano 1–7 dni po zakończeniu radioterapii. Odsetek ostrych powikłań popromiennych i ich nasilanie było umiarkowane. Niską resekcję przednią wykonano u 24 (51%) chorych, amputację odbytnicy sposobem brzuszno-kroczowym u 19 (40%), operację Hartmannu u 2 (4%) i (74%) chorych w podgrupie, w której nowotwór nie naciekał zwieraczy. Usuwano całe mezorektum. Margines pomiędzy dolnym brzegiem makroskopowo widocznego guza, a cięciem chirurgicznym jelita wynosił od 0,1 cm do 3 cm, mediana 0,8 cm (nie uwzględniono krążków ze staplera).Wszyscy chorzy przeżyli operację 30-dniowy okres pooperacyjny. Odsetek powikłań pooperacyjnych nie odbiegał znacząco od obserwowanych po porównywalnych operacjach u chorych nienapromienianych. Uzupełniającą chemioterapię otrzymało 20 chorych (43%). Okres obserwacji wynosi od 7 miesięcy do 30 miesięcy, mediana 15 miesięcy. Wznowę miejscową obserwowano w dwóch przypadkach (8%); późne powikłania popromienne pod postacią bólów w okolicy krzyżowej u dwóch chorych (8%).Wczesne wyniki sugerują wysoką miejscową skuteczność leczenia przy stosunkowo niewielkim odsetku powikłań i znacznym odsetku chorych z zachowanymi zwieraczami

Pkd1 Regulates Lymphatic Vascular Morphogenesis during Development.

Lymphatic vessels arise during development through sprouting of precursor cells from veins, which is regulated by known signaling and transcriptional mechanisms. The ongoing elaboration of vessels to form a network is less well understood. This involves cell polarization, coordinated migration, adhesion, mixing, regression, and shape rearrangements. We identified a zebrafish mutant, lymphatic and cardiac defects 1 (lyc1), with reduced lymphatic vessel development. A mutation in polycystic kidney disease 1a was responsible for the phenotype. PKD1 is the most frequently mutated gene in autosomal dominant polycystic kidney disease (ADPKD). Initial lymphatic precursor sprouting is normal in lyc1 mutants, but ongoing migration fails. Loss of Pkd1 in mice has no effect on precursor sprouting but leads to failed morphogenesis of the subcutaneous lymphatic network. Individual lymphatic endothelial cells display defective polarity, elongation, and adherens junctions. This work identifies a highly selective and unexpected role for Pkd1 in lymphatic vessel morphogenesis during development

New International Guidelines and Consensus on the Use of Lung Ultrasound

Following the innovations and new discoveries of the last 10 years in the field of lung ultrasound (LUS), a multidisciplinary panel of international LUS experts from six countries and from different fields (clinical and technical) reviewed and updated the original international consensus for point-of-care LUS, dated 2012. As a result, a total of 20 statements have been produced. Each statement is complemented by guidelines and future developments proposals. The statements are furthermore classified based on their nature as technical (5), clinical (11), educational (3), and safety (1) statements

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

Background Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide.Methods A multimethods analysis was performed as part of the GlobalSurg 3 study-a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital.Findings Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3.85 [95% CI 2.58-5.75]; p<0.0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63.0% vs 82.7%; OR 0.35 [0.23-0.53]; p<0.0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer.Interpretation Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised

Semaglutide and cardiovascular outcomes in patients with obesity and prevalent heart failure: a prespecified analysis of the SELECT trial

Background: Semaglutide, a GLP-1 receptor agonist, reduces the risk of major adverse cardiovascular events (MACE) in people with overweight or obesity, but the effects of this drug on outcomes in patients with atherosclerotic cardiovascular disease and heart failure are unknown. We report a prespecified analysis of the effect of once-weekly subcutaneous semaglutide 2·4 mg on ischaemic and heart failure cardiovascular outcomes. We aimed to investigate if semaglutide was beneficial in patients with atherosclerotic cardiovascular disease with a history of heart failure compared with placebo; if there was a difference in outcome in patients designated as having heart failure with preserved ejection fraction compared with heart failure with reduced ejection fraction; and if the efficacy and safety of semaglutide in patients with heart failure was related to baseline characteristics or subtype of heart failure. Methods: The SELECT trial was a randomised, double-blind, multicentre, placebo-controlled, event-driven phase 3 trial in 41 countries. Adults aged 45 years and older, with a BMI of 27 kg/m2 or greater and established cardiovascular disease were eligible for the study. Patients were randomly assigned (1:1) with a block size of four using an interactive web response system in a double-blind manner to escalating doses of once-weekly subcutaneous semaglutide over 16 weeks to a target dose of 2·4 mg, or placebo. In a prespecified analysis, we examined the effect of semaglutide compared with placebo in patients with and without a history of heart failure at enrolment, subclassified as heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, or unclassified heart failure. Endpoints comprised MACE (a composite of non-fatal myocardial infarction, non-fatal stroke, and cardiovascular death); a composite heart failure outcome (cardiovascular death or hospitalisation or urgent hospital visit for heart failure); cardiovascular death; and all-cause death. The study is registered with ClinicalTrials.gov, NCT03574597. Findings: Between Oct 31, 2018, and March 31, 2021, 17 604 patients with a mean age of 61·6 years (SD 8·9) and a mean BMI of 33·4 kg/m2 (5·0) were randomly assigned to receive semaglutide (8803 [50·0%] patients) or placebo (8801 [50·0%] patients). 4286 (24·3%) of 17 604 patients had a history of investigator-defined heart failure at enrolment: 2273 (53·0%) of 4286 patients had heart failure with preserved ejection fraction, 1347 (31·4%) had heart failure with reduced ejection fraction, and 666 (15·5%) had unclassified heart failure. Baseline characteristics were similar between patients with and without heart failure. Patients with heart failure had a higher incidence of clinical events. Semaglutide improved all outcome measures in patients with heart failure at random assignment compared with those without heart failure (hazard ratio [HR] 0·72, 95% CI 0·60-0·87 for MACE; 0·79, 0·64-0·98 for the heart failure composite endpoint; 0·76, 0·59-0·97 for cardiovascular death; and 0·81, 0·66-1·00 for all-cause death; all pinteraction>0·19). Treatment with semaglutide resulted in improved outcomes in both the heart failure with reduced ejection fraction (HR 0·65, 95% CI 0·49-0·87 for MACE; 0·79, 0·58-1·08 for the composite heart failure endpoint) and heart failure with preserved ejection fraction groups (0·69, 0·51-0·91 for MACE; 0·75, 0·52-1·07 for the composite heart failure endpoint), although patients with heart failure with reduced ejection fraction had higher absolute event rates than those with heart failure with preserved ejection fraction. For MACE and the heart failure composite, there were no significant differences in benefits across baseline age, sex, BMI, New York Heart Association status, and diuretic use. Serious adverse events were less frequent with semaglutide versus placebo, regardless of heart failure subtype. Interpretation: In patients with atherosclerotic cardiovascular diease and overweight or obesity, treatment with semaglutide 2·4 mg reduced MACE and composite heart failure endpoints compared with placebo in those with and without clinical heart failure, regardless of heart failure subtype. Our findings could facilitate prescribing and result in improved clinical outcomes for this patient group. Funding: Novo Nordisk