Modelling the age distribution of longevity leaders.

Human longevity leaders with remarkably long lifespan play a crucial role in the advancement of longevity research. In this paper, we propose a stochastic model to describe the evolution of the age of the oldest person in the world by a Markov process, in which we assume that the births of the individuals follow a Poisson process with increasing intensity, lifespans of individuals are independent and can be characterized by a gamma-Gompertz distribution with time-dependent parameters. We utilize a dataset of the world's oldest person title holders since 1955, and we compute the maximum likelihood estimate for the parameters iteratively by numerical integration.

How job crafting is related to the individual readiness to organizational change.

This article aims to examine the relationship between job crafting activities and employees' readiness to change. Confirmatory factor analysis and hierarchical regression analysis were conducted on a representative sample of 500 employees. Sampling was carried out in a European country in a period strongly affected by COVID-19 to isolate the five dimensions of job crafting and their separate effects on employees' readiness to change.

On the Technology Acceptance Behavior of Romanian Preschool Teachers.

Unlabelled: This study investigates how compatibility and perceived enjoyment affect the link between intention to use and actual technology use in Romanian preschool education, building on earlier studies.

Methods: 300 participants were invited to participate in this research from 15 Romanian counties. 182 preschool teachers' questionnaires were utilized for data analysis after the return and screening of responses.

The use of theory of planned behavior to systemically study the integrative-qualitative intentional behavior in Romanian preschool education with network analysis.

Predicting preschool teachers' intention to adopt qualitative and inclusive early childhood intentional behaviors represents an important research field. The objective of this research is first to develop and validate a scale to assess the integrative-qualitative intentional behavior (IQIB-ECEC) of preschool teachers in order to achieve SDG4.2's objective of ensuring that all children have access to high-quality pre-primary education and then to systemically analyze the relationship between variables with Network Analysis.

Pathways to inclusive and equitable quality early childhood education for achieving SDG4 goal-a scoping review.

According to Sustainable Development Goal 4.2 (SDG 4.2), Equal Access to Quality Pre-primary Education, governments throughout the world are working to ensure that all children have access to high-quality early childhood development, care, and pre-primary education by 2030.

Cryopreservation of gander semen in cryovials - Comparative study.

The aim of the study was to find a practical and inexpensive method for freezing goose semen for use in routine inseminations under farm conditions. Two basic freezing protocols [(1) dynamic, programmable freezing and (2) static, nitrogen vapour method] were evaluated with varying concentrations of dimethylformamide (DMF) plus additional osmoprotectants such as betaine, trehalose, and sucrose, using cryovials as containers. Altogether eight different treatments were compared.

Evolution and characterization of a new reversibly photoswitching chromogenic protein, Dathail.

We report the engineering of a new reversibly switching chromogenic protein, Dathail. Dathail was evolved from the extremely thermostable fluorescent proteins thermal green protein (TGP) and eCGP123 using directed evolution and ratiometric sorting. Dathail has two spectrally distinct chromogenic states with low quantum yields, corresponding to absorbance in a ground state with a maximum at 389nm, and a photo-induced metastable state with a maximum at 497nm.

Recombinant renewable polyclonal antibodies.

Only a small fraction of the antibodies in a traditional polyclonal antibody mixture recognize the target of interest, frequently resulting in undesirable polyreactivity. Here, we show that high-quality recombinant polyclonals, in which hundreds of different antibodies are all directed toward a target of interest, can be easily generated in vitro by combining phage and yeast display. We show that, unlike traditional polyclonals, which are limited resources, recombinant polyclonal antibodies can be amplified over one hundred million-fold without losing representation or functionality.

From deep sequencing to actual clones.

The application of deep sequencing to in vitro display technologies has been invaluable for the straightforward analysis of enriched clones. After sequencing in vitro selected populations, clones are binned into identical or similar groups and ordered by abundance, allowing identification of those that are most enriched. However, the greatest strength of deep sequencing is also its greatest weakness: clones are easily identified by their DNA sequences, but are not physically available for testing without a laborious multistep process involving several rounds of polymerization chain reaction (PCR), assembly and cloning.

Incidence and molecular diversity of Acanthamoeba species isolated from public baths in Hungary.

Hungary has a large number of thermal baths and spa facilities which attract hundreds of thousands of tourists annually. Until recently, however, the free-living amoebae were not of public health concern. Genotyping of Acanthamoeba species, potential agents of keratitis and granulomatous encephalitis, was carried out in 20 Hungarian public baths for the first time to assess the incidence and molecular diversity of the genus in the country.

The antibody mining toolbox: an open source tool for the rapid analysis of antibody repertoires.

In vitro selection has been an essential tool in the development of recombinant antibodies against various antigen targets. Deep sequencing has recently been gaining ground as an alternative and valuable method to analyze such antibody selections. The analysis provides a novel and extremely detailed view of selected antibody populations, and allows the identification of specific antibodies using only sequencing data, potentially eliminating the need for expensive and laborious low-throughput screening methods such as enzyme-linked immunosorbant assay.

Using phage display selected antibodies to dissect microbiomes for complete de novo genome sequencing of low abundance microbes.

Background: Single cell genomics has revolutionized microbial sequencing, but complete coverage of genomes in complex microbiomes is imperfect due to enormous variation in organismal abundance and amplification bias. Empirical methods that complement rapidly improving bioinformatic tools will improve characterization of microbiomes and facilitate better genome coverage for low abundance microbes.

Methods: We describe a new approach to sequencing individual species from microbiomes that combines antibody phage display against intact bacteria with fluorescence activated cell sorting (FACS).

Phanta: a non-fluorescent photochromic acceptor for pcFRET.

We have developed an orange non-fluorescent photochromic protein (quantum yield, 0.003) we call Phanta that is useful as an acceptor in pcFRET applications. Phanta can be repeatedly inter-converted between the two absorbing states by alternate exposure to cyan and violet light.

Specific binder for Lightning-Link® biotinylated proteins from an antibody phage library.

Many applications required protein biotinylation. We routinely use biotinylated proteins to select single chain antibodies from phage and/or yeast display libraries. During phage selection the biotinylated antigens are bound to streptavidin coupled magnetic beads, while during yeast display, the biotinylated antigens are used during flow cytometry for both analysis and sorting.

Time-resolved, confocal single-molecule tracking of individual organic dyes and fluorescent proteins in three dimensions.

We demonstrate following individual fluorescent protein constructs and individual organic dyes as they diffuse in 3-D in solution at rates up to 1 μm(2)/s over distances of several micrometers in X, Y, and Z. Our 3-D tracking method is essentially a stage scanning confocal microscope that uses a unique spatial filter geometry and active feedback 200 times/s to follow fast 3-D motion. Here we detail simulations used to find optimal feedback parameters for following individual fluorescent proteins in 3-D and show that a wide range of parameters are capable of following individual proteins diffusing at 1 μm(2)/s rates.

Expression, purification, crystallization and preliminary X-ray analysis of eCGP123, an extremely stable monomeric green fluorescent protein with reversible photoswitching properties.

Enhanced consensus green protein variant 123 (eCGP123) is an extremely thermostable green fluorescent protein (GFP) that exhibits useful negative reversible photoswitching properties. eCGP123 was derived by the application of both a consensus engineering approach and a recursive evolutionary process. Diffraction-quality crystals of recombinant eCGP123 were obtained by the hanging-drop vapour-diffusion method using PEG 3350 as the precipitant.

[Systemic autoimmune disorders and pregnancy].

The coincidence of systemic autoimmune diseases and pregnancy may modify the outcome of the disease and the pregnancy due to the background immunologic and hormonal processes. The great majority of patients with autoimmune diseases are young females in their reproductive years, willing to have babies. Consequently, we have to prepare for this special situation.

"To be or not to be," ten years after: evidence for mixed connective tissue disease as a distinct entity.

Objectives: To determine if mixed connective tissue disease (MCTD) can be considered an independent clinical entity, to compare 3 different classification criteria for MCTD (Kasukawa, Alarcón-Segovia, and Sharp), and to define predictors (clinical features and autoantibodies) of potential evolution toward other connective tissue diseases (CTDs).

Methods: One hundred sixty-one MCTD patients were evaluated retrospectively at the diagnosis and in 2008. They were classified, at the diagnosis, according to the 3 classification criteria of MCTD (Sharp, Alarcón-Segovia, and Kasukawa) and reclassified in 2008 according to their evolution.

Filtering "genic" open reading frames from genomic DNA samples for advanced annotation.

Background: In order to carry out experimental gene annotation, DNA encoding open reading frames (ORFs) derived from real genes (termed "genic") in the correct frame is required. When genes are correctly assigned, isolation of genic DNA for functional annotation can be carried out by PCR. However, not all genes are correctly assigned, and even when correctly assigned, gene products are often incorrectly folded when expressed in heterologous hosts.

A comprehensive analysis of filamentous phage display vectors for cytoplasmic proteins: an analysis with different fluorescent proteins.

Filamentous phage display has been extensively used to select proteins with binding properties of specific interest. Although many different display platforms using filamentous phage have been described, no comprehensive comparison of their abilities to display similar proteins has been conducted. This is particularly important for the display of cytoplasmic proteins, which are often poorly displayed with standard filamentous phage vectors.

Directed evolution of an extremely stable fluorescent protein.

In this paper we describe the evolution of eCGP123, an extremely stable green fluorescent protein based on a previously described fluorescent protein created by consensus engineering (CGP: consensus green protein). eCGP123 could not be denatured by a standard thermal melt, preserved almost full fluorescence after overnight incubation at 80 degrees C and possessed a free energy of denaturation of 12.4 kcal/mol.

Protein tyrosine phosphatase gene C1858T allele confers risk for rheumatoid arthritis in Hungarian subjects.

The C1858T allele of the PTPN22 gene has been reported to confer risk for RA; but in some reports, the effect was restricted to RF- and/or anti-CCP-seropositive patients. Hungarian RA patients and matched controls were genotyped. The 1858T allele showed an increased prevalence in RA patients compared to controls.