Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells

This protocol measures the 3D Euclidean distance (Δ3D) between two/three fluorescently labeled kinetochore components in fixed samples using Kinetochore Delta software (KiDv1.0.1, MATLAB based). Overestimation of mean Δ3D is corrected through a Bayesian algorithm, with ΔEC distances reflecting the ensemble average positions of fluorophores within a kinetochore population. This package also enables kinetochore categorization, which can be used to sub-sample kinetochores and measure ΔEC. Together, this allows the dynamic architecture of human kinetochores to be investigated (tested in hTERT-RPE1 cells)

Bayesian inference of multi-point macromolecular architecture mixtures at nanometre resolution

Gaussian spot fitting methods have significantly extended the spatial range where fluorescent microscopy can be used, with recent techniques approaching nanometre (nm) resolutions. However, small inter-fluorophore distances are systematically over-estimated for typical molecular scales. This bias can be corrected computationally, but current algorithms are limited to correcting distances between pairs of fluorophores. Here we present a flexible Bayesian computational approach that infers the distances and angles between multiple fluorophores and has several advantages over these previous methods. Specifically it improves confidence intervals for small lengths, estimates measurement errors of each fluorophore individually and infers the correlations between polygon lengths. The latter is essential for determining the full multi-fluorophore 3D architecture. We further developed the algorithm to infer the mixture composition of a heterogeneous population of multiple polygon states. We use our algorithm to analyse the 3D architecture of the human kinetochore, a macro-molecular complex that is essential for high fidelity chromosome segregation during cell division. Using triple fluorophore image data we unravel the mixture of kinetochore states during human mitosis, inferring the conformation of microtubule attached and unattached kinetochores and their proportions across mitosis. We demonstrate that the attachment conformation correlates with intersister tension and sister alignment to the metaphase plate

Liver grafts procured and discarded by all Belgian centers and transplanted within Eurotransplant network: analysis of cause to decline, a Be-LIAC study

The annual balance between imports and exports of grafts is a matter of debate. We examined the Eurotransplant database of all liver grafts procured within Belgium and Luxemburg which were exported and transplanted abroad. The aim of our study was to analyse the reasons for graft refusal by all Belgian transplant centres and early postoperative evolution. Database between 2015 and 2019 included donor characteristics, reason of offer decline, graft and recipient survival. During the 4 year period 329 grafts were procured in Belgium and transplanted abroad. 163 were exported for HU recipients, 17 no national match recipients (8 AB group, 2 pediatric and 7 other reasons), 19 pay back, 15 splits, 11 not mentioned. Hundred and four grafts were declined by all Belgian centres. Forty seven were declined primary offers and fifty seven livers were distributed by extended allocation. Between them we find out four DCD donors, 83 for medical reasons (age, cytolysis, size mismatch and steatosis). Thirteen livers were accepted and declined at arrival for size mismatch (kept as rescue offer in the same centre). One donor was unstable and two were rejected for positive HCV virology. Only one liver who was primary accepted for a split was transplanted as a whole liver. Two patients presented primary graft nonfunction and three primary graft dysfunction. All of them were retransplanted. Thirteen patients died in the early 3 month postoperative period. Even though higher mortality is expected from marginal grafts, better acceptance rate could be achieved at a national level

Brucellosis: Trends in the Development of Situation in the World and Forecast for 2022 in the Russian Federation

An analysis of trends in the development of situation on brucellosis in the world over past decade and the data on the main risk factors for the occurrence of epidemiological complications regarding this infection in various regions of the world are provided in the paper. An expert assessment of the current epizootiological and epidemiological situation on brucellosis, the coverage of population and animals with immunization in the Russian Federation is given. Over 9 months of 2021, 210 potentially hazardous as regards brucellosis in cattle areas and 24 sites – as regards brucellosis in small ruminants – were registered in Russia. Compared to the same period in 2020, there was a decrease in the number of newly identified hazardous sites for bovine brucellosis by 35.8 % (117 areas). However, long-term upward trend in epizootiological adversity for bovine brucellosis in Russia persists. The epidemiological situation on brucellosis in the country for the period of 2012–2021 is characterized as unfavorable. Decrease in the number of newly detected human brucellosis cases (by 25.1 % of long-term average values) is observed against the background of persistent unfavorable epizootic conditions for brucellosis among epidemiologically significant species of small ruminants and cattle in regions with developed animal husbandry. In 2021, clusters of human cases were registered in the Republic of Dagestan and Penza Region. In the Republic of Dagestan, against the background of aggravation of epizootiological and epidemiological situation on brucellosis, there was also an alarming trend towards prevalence of a relatively high incidence among minors. The proportion of cases of brucellosis among children under the age of 17 in the Republic amounted to 60.3 % of the total number of minors with newly diagnosed brucellosis in Russia over the past 10 years. Taking into account current epizootic, epidemic situations and the long-term dynamics of the development of situation on  brucellosis in the Russian Federation, the incidence of brucellosis among the population  is predicted to be 10–15 % lower than the average long-term values – 0.18–0.20 per 100000 of the population – in 2022. The number of human cases of brucellosis can range from 250 to 300

Analysis of Brucellosis Incidence and Molecular-Genetic Characteristics of Brucella Population in the Territory of the Russian Federation

An analysis of brucellosis incidence in Russia in 2013–2022 and the data on genetic diversity of Brucella melitensis and Brucella abortus populations isolated in Russia in 1939–2022 are provided in the review. Over the past decade, the epidemiological situation in Russia has been characterized as unstable against the background of persistent unfavorable conditions for brucellosis in cattle and small ruminants. During the period of 2013–2022 (9 months), 4298 epizootic foci as regards brucellosis in cattle (89164 sick animals) and 371 as regards brucellosis in small ruminants (13569) were registered. The largest number of epizootic brucellosis foci was recorded in the North-Caucasian and Southern Federal Districts. In 2013–2022, on average, 327 cases of brucellosis among people were registered annually, the incidence rate per 100 000 of the population was 0.24. Up to 70–90 % of brucellosis cases were detected in the south of the European part of the country. A trend towards deterioration of the situation on brucellosis in Volga (Penza and Samara Regions) and Central (Smolensk, Voronezh and Tula Regions) Federal Districts is observed. There is a connection between the intensity of epidemic manifestations of brucellosis and the level of anthropurgic enzooty of territories. In 2022, 467 cases of brucellosis were reported (0.32 per 100 000 of the population), which is 42.8 % higher than annual average values over 10 years. In 2023, a measure of stability of incidence rates, by 20–25 % above average long-term values, is to be expected. Incidence of brucellosis in humans will be within the range of 380–410 cases (0.26–0.28 per 100 000 of the population). The results of genotyping of B. melitensis strains point to an increase in the proportion of isolates with an MLVA-profile characteristic of strains from enzootic as regards brucellosis countries of Middle East and North Africa over past 20–25 years, which can indicate importation (introduction) of the infection from these territories to Russia through small ruminants and/or biomaterial from them

A multicentre outcome analysis to define global benchmarks for donation after circulatory death liver transplantation

Cellular mechanisms in basic and clinical gastroenterology and hepatolog

Hemorrhoidectomy using harmonic ultrasound scalpel : results of a series of 110 consecutive patients

As a very common and benign disease in surgical practice, hemorhoidal disease should have a very low level of postoperative morbidity and pain. The aim of this study was to evaluate postoperative pain and morbidity, time to complete healing and time off of work in a series of 110 consecutive hemorrhoidectomy patients using harmonic ultrasound scalpel device. One-hundred-and-ten consecutive three quadrant hemorrhoidectomy procedures for symptomatic prolapsed hemorrhoids (grade III and IV) were performed in our institution. Postoperative pain was assessed using a visual analogue pain scale (VAS). Outpatient`s follow-up was done at 3 weeks and 3 months. There were 72 males (65%) and 38 females (35%). Mean age was 48,5 years (22-76). Sixty-seven patients (61%) had grade III hemorrhoids and 43 patients (39%), grade IV. All operations were performed under spinal anaesthesia. Postoperative complications included 7 patients (6,3%): 2 patients were readmitted for haemorrhage without need for reintervention, and 5 had urinary retention. Postoperatively scheduled analgesia was enough for pain control in all patients. One-day surgery represents only 5% of our patients, 85% had a less than 24 hours hospital stay with surgery done the day of admission and 10% stayed for more than one day. Seventy-nine patients (92%) returned to work in between 3 weeks. Time to complete healing was 6 weeks. At the term of the follow-up, no recurrence was observed. In conclusion, hemorroidectomy using harmonic scalpel is a safe technique with very low morbidity rate, postoperative pain is acceptable and time off of work is low