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IntroductionThe two-component signal transduction systems play an essential role in the adaptation of bacteria to changing environmental conditions. One of them is the MnoSR system involved in the regulation of methylotrophic metabolism in M. smegmatis.MethodsMycobacterium smegmatis mutant strains ΔmnoS, ΔmnoR and ΔmnoS/R lacking functional mnoS, mnoR and both genes were generated using a homologous recombination approach. MnoR recombinant protein was purified by affinity column chromatography. The present study employs molecular biology techniques: cloning strategies, global RNA sequencing, qRT-PCR, EMSA, Microscale thermophoresis, and bioinformatics analysis.Results and discussionThe ∆mnoS, ∆mnoR, and ∆mnoS/R mutant strains were generated and cultured in the presence of defined carbon sources. Growth curve analysis confirmed that inactivation of the MnoSR impairs the ability of M. smegmatis cells to use alcohols such as 1,3-propanediol and ethanol but improves the bacterial growth on ethylene glycol, xylitol, and glycerol. The total RNA sequencing method was employed to understand the importance of MnoSR in the global responses of mycobacteria to limited carbon access and in carbon-rich conditions. The loss of MnoSR significantly affected carbon utilization in the case of mycobacteria cultured on glucose or 1,3-propanediol as sole carbon sources as it influenced the expression of multiple metabolic pathways. The numerous transcriptional changes could not be linked to the presence of evident MnoR DNA-binding sites within the promotor regions for the genes outside of the mno operon. This was confirmed by EMSA and microscale thermophoresis with mutated MnoR binding consensus region. Our comprehensive analysis highlights the system’s vital role in metabolic adaptability, providing insights into its potential impact on the environmental survival of mycobacteria.

Taurine is the most abundant non-proteinogenic sulphur amino acid in humans, and its level decreases dramatically with aging. In humans, the highest levels of taurine are present in retina, leukocytes, heart, skeletal muscles and brain. Taurine is necessary to ensure proper homeostasis and recent research in animals (for example mice and monkeys) have shown that it significantly increases life expectancy and improves health. At the cellular level, taurine is involved in a number of processes such as osmoregulation, modulation of calcium levels and stabilization of cell membranes. Moreover, it has an inhibitory effect on the central nervous system and has antioxidant, anti-apoptotic and anti-inflammatory activity. The development of neurodegenerative diseases is associated with improper functioning of mitochondria, oxidative stress, development of inflammation, excessive activation of ionotropic and metabotropic glutamate receptors and neuronal apoptosis. In vitro and in vivo studies have shown that taurine inhibits the above-mentioned pathological processes and has neuroprotective properties. The antioxidant effect of taurine results from neutralizing hypochlorous acid, a strong oxidant that is produced by neutrophils at the site of the inflammation. Taurine chloramine (Tau-Cl) produced in this reaction reduces oxidative stress and inhibits the production of pro-inflammatory cytokines (e.g. TNF-α, IL-1β, IL-6), nitric oxide (NO) and prostaglandin E2. Moreover, this compound stimulates the expression of antioxidant enzymes (e.g. peroxiredoxin 1, thioredoxin-1, hemooxygenase-1, glutathione peroxidase, catalase). Within the nervous tissue, the anti-inflammatory and antioxidant effect of taurine is related to reduced expression of IL-1α, IL-1β, TNF-α, IFN-γ, IL-6 and GM-CSF, and increase in the activity of antioxidant enzymes (peroxidase-, reductase- and S- glutathione transferase, superoxide dismutase) and glutathione concentration. Taurine is necessary for the proper functioning of mitochondria. Its deficiency blocks the synthesis of taurine-modified leucine tRNA (τm5U-tRNALeu), which results in inhibition of the synthesis of NADH-ubiquinone oxidoreductase, improper functioning of complex I of the respiratory chain and increase in oxidative stress. Endoplasmic reticulum (ER) stress is a hallmark of neurodegenerative diseases and can lead to neuronal apoptosis. Taurine deficiency increases the accumulation of misfolded cellular proteins and induces ER stress. Taurine supplementation reduces neuronal ER stress by inhibiting ATF6- and IRE-1-dependent signalling pathways. Moreover, taurine inhibits apoptosis of neuronal cells by increasing the expression of Bcl-2, decreasing the expression of p53, CHOP and Bax and reducing the activity of caspase-3. In the central nervous system, taurine also serves as an inhibitory neurotransmitter due to binding with γ-aminobutyric acid and glycine receptors (GABAA and GlyR, respectively). Such interactions causes an increased influx of chloride ions into neurons, hyperpolarization of the cell membrane and, consequently, inhibition of glutamatergic neurotransmission and excitotoxicity induced by glutamic acid. The neuroprotective activity of taurine is due to its pleiotropic effects related to the inhibition of apoptosis, oxidative stress, ER stress, and also to its anti-inflammatory activity and inhibition of neuronal excitability.

Geodashboards are often designed with explanatory elements guiding users. These elements (e.g. legends or annotations) need to be carefully designed to mitigate split attention or information integration issues. In this paper, we report expert interviews followed by a controlled experiment where we compare two interface designs with a focus on the split attention effect: (1) a multiple-legend layout with explanatory elements located next to each view, and (2) a single-legend layout with all explanatory elements gathered in one place. Different legend layouts did not affect the performance, but affected user satisfaction. 75% of the participants preferred the multiple-legend layout, and rated it with a higher usability score, mainly attributing this preference to the proximity of legend elements to the view of interest. Eye tracking data strongly and clearly verifies that participants indeed make use of the proximity: With the single-legend, the majority of eye-movement transitions were between the single-legend and the closest view to the legend, whereas with multiple-legend participants have shorter and more frequent legend visits, as well as more transitions between legends and views. Taken together, the design lesson we learned from this experiment can be summarized as ‘split the legend elements, but make it close to the explained elements’.

Abstract In reference to gene annotation, more than half of the tRNA species synthesized by Mycobacterium tuberculosis require the enzymatic addition of the cytosine-cytosine-adenine (CCA) tail, which is indispensable for amino acid charging and tRNA functionality. It makes the mycobacterial CCA-adding enzyme essential for survival of the bacterium and a potential target for novel pipelines in drug discovery avenues. Here, we described the rv3907c gene product, originally annotated as poly(A)polymerase (rv3907c, PcnA) as a functional CCA-adding enzyme (CCA Mtb ) essential for viability of M. tuberculosis. The depletion of the enzyme affected tRNAs maturation, inhibited bacilli growth, and resulted in abundant accumulation of polyadenylated RNAs. We determined the enzymatic activities displayed by the mycobacterial CCA Mtb in vitro and studied the effects of inhibiting of its transcription in bacterial cells. We are the first to properly confirm the existence of RNA polyadenylation in mycobacteria, a previously controversial phenomenon, which we found promoted upon CCA-adding enzyme downexpression.

Chitosan is a very promising material for tissue model printing. It is also known that the introduction of chemical modifications to the structure of the material in the form of methacrylate groups makes it very attractive for application in the bioprinting of tissue models. The aim of this work is to study the characteristics of biomaterials containing chitosan (BCH) and its methacrylated equivalent (BCM) in order to identify differences in their usefulness in 3D bioprinting technology. It has been shown that the BCM material containing methacrylic chitosan is three times more viscous than its non-methacrylated BCH counterpart. Additionally, the BCM material is characterized by stability in a larger range of stresses, as well as better printability, resolution, and fiber stability. The BCM material has higher mechanical parameters, both mechanical strength and Young’s modulus, than the BCH material. Both materials are ideal for bioprinting, but BCM has unique rheological properties and significant mechanical resistance. In addition, biological tests have shown that the addition of chitosan to biomaterials increases cell proliferation, particularly in 3D-printed models. Moreover, modification in the form of methacrylation encourages reduced toxicity of the biomaterial in 3D constructs. Our investigation demonstrates the suitability of a chitosan-enhanced biomaterial, specifically methacrylate-treated, for application in tissue engineering, and particularly for tissues requiring resistance to high stress, i.e., vascular or cartilage models.

The recombinant structural protein described in this study was designed based on sequences derived from elastin and silk. Silk–elastin hybrid copolymers are characterized by high solubility while maintaining high product flexibility. The phase transition temperature from aqueous solution to hydrogel, as well as other physicochemical and mechanical properties of such particles, can differ significantly depending on the number of sequence repeats. We present a preliminary characterization of the EJ17zipR protein obtained in high yield in a prokaryotic expression system and efficiently purified via a multistep process. Its addition significantly improves biomaterial’s rheological and mechanical properties, especially elasticity. As a result, EJ17zipR appears to be a promising component for bioinks designed to print spatially complex structures that positively influence both shape retention and the internal transport of body fluids. The results of biological studies indicate that the addition of the studied protein creates a favorable microenvironment for cell adhesion, growth, and migration.

In this study we propose to use for bioprinting a bioink enriched with a recombinant RE15mR protein with a molecular weight of 26 kDa, containing functional sequences derived from resilin and elastin. The resulting protein also contains RGD sequences in its structure, as well as a metalloproteinase cleavage site, allowing positive interaction with the cells seeded on the construct and remodeling the structure of this protein in situ. The described protein is produced in a prokaryotic expression system using an E. coli bacterial strain and purified by a process using a unique combination of known methods not previously used for recombinant elastin-like proteins. The positive effect of RE15mR on the mechanical, physico-chemical, and biological properties of the print is shown in the attached results. The addition of RE15mR to the bioink resulted in improved mechanical and physicochemical properties and promoted the habitation of the prints by cells of the L-929 line.

Currently, a major challenge in material engineering is to develop a cell-safe biomaterial with significant utility in processing technology such as 3D bioprinting. The main goal of this work was to optimize the composition of a new graphene oxide (GO)-based bioink containing additional extracellular matrix (ECM) with unique properties that may find application in 3D bioprinting of biomimetic scaffolds. The experimental work evaluated functional properties such as viscosity and complex modulus, printability, mechanical strength, elasticity, degradation and absorbability, as well as biological properties such as cytotoxicity and cell response after exposure to a biomaterial. The findings demonstrated that the inclusion of GO had no substantial impact on the rheological properties and printability, but it did enhance the mechanical properties. This enhancement is crucial for the advancement of 3D scaffolds that are resilient to deformation and promote their utilization in tissue engineering investigations. Furthermore, GO-based hydrogels exhibited much greater swelling, absorbability and degradation compared to non-GO-based bioink. Additionally, these biomaterials showed lower cytotoxicity. Due to its properties, it is recommended to use bioink containing GO for bioprinting functional tissue models with the vascular system, e.g., for testing drugs or hard tissue models.

The genetic factors of adult acute lymphoblastic leukemia (ALL) development are only partially understood. The Runt-Related Transcription Factor (RUNX) gene family play a crucial role in hematological malignancies, serving both a tumor suppressor and promoter function. The aim of this study was the assessment of relative RUNX1 and RUNX3 genes expression level among adult ALL cases and a geographically and ethnically matched control group. The relative RUNX1 and RUNX3 genes expression level was assessed by qPCR. The investigated group comprised 60 adult patients newly diagnosed with ALL. The obtained results were compared with a group of 40 healthy individuals, as well as clinical and hematological parameters of patients, and submitted for statistical analysis. ALL patients tend to have significantly higher RUNX1 gene expression level compared with controls. This observation is also true for risk group stratification where high-risk (HR) patients presented higher levels of RUNX1. A higher RUNX1 transcript level correlates with greater leukocytosis while RUNX3 expression is reduced in Philadelphia chromosome bearers. The conducted study sustains the hypothesis that both a reduction and increase in the transcript level of RUNX family genes may be involved in leukemia pathogenesis, although their interaction is complex. In this context, overexpression of the RUNX1 gene in adult ALL cases in particular seems interesting. Obtained results should be interpreted with caution. Further analysis in this research field is needed.

This study aims to analyze the ceramic-metal composite Al2O3/TiO2/TiAl2O5 obtained using the slip-casting method. Samples containing 50% vol. of the solid phase and 2% vol. and 4% vol. fractions of the metallic phase were examined. Rheological investigations were performed. Measurements of shrinkage and density of the composites produced were determined. The phase composition of the obtained composite was investigated using SEM/EDS and XRD techniques. Stereological analysis was performed as well. The slip-casting method enables the production of the proposed composite, reinforced by the presence of TiO2 and TiAl2O5. With the increase in the content of the metallic phase in the composite, the thialite phase content increases, but relative density and volumetric shrinkage of the obtained composites both decrease. Thialite grains are characterized by a size in the range of 4 µm to 15 µm, which leads to a low density of the samples. The results revealed that no significant effect of changing the metal phase content of the slurries used for the composites being fabricated was observed on the limiting grain growth of alumina during the sintering process of slip-casting composites. This finding is important as it suggests that the increase in metallic phase content does not lead to undesirable grain coarsening, which could degrade mechanical properties. [ABSTRACT FROM AUTHOR]

Objectives: The aim of this study was to evaluate which anthropometric index, either body mass index (BMI) or weight-adjusted waist index (WWI), is more accurately associated with the prevalence of the most common chronic diseases and components of geriatric assessment in hospitalized older adults. Methods: The study included a total of 2945 hospitalized older adults (median age 82 years). The associations between the presence of chronic diseases and Comprehensive Geriatric Assessment (CGA) results were compared with WWI and BMI values. Results: The WWI was significantly higher in both sex groups suffering from hypertension, diabetes, osteoarthritis, and depression. In women, the parameter was increased among individuals with previous myocardial infarction, who presented heart failure symptoms or had chronic kidney disease diagnosed, whereas in men, among those with pulmonary diseases and osteoporosis, WWI was related to many CGA parameters oftentimes where BMI proved to fail. There was a positive correlation of WWI with the presence of depressive symptoms assessed with the geriatric depression scale (GDS) but no significant correlation with BMI. In multiple logistic regression models, WWI was a stronger predictor of depression as compared to waist circumference or the waist-to-height ratio. Conclusions: There is an association between a higher WWI and depression diagnosis as well as the presence of depressive symptoms according to the GDS in hospitalized older adults, both women and men. There is no such correlation between depression and BMI. Both high BMI and high WWI values seem to identify older patients with cardiometabolic diseases such as hypertension and diabetes. According to this study, WWI seems to be a promising indicator of depression risk and, similarly to BMI, a useful parameter for the assessment of cardiometabolic risk in older hospitalized adults. [ABSTRACT FROM AUTHOR]

It was recently reported that 4-substituted picolinohydrazonamides carrying hydrophilic cyclic amines, such as morpholine and pyrrolidine, at the end of their thiosemicarbazide chain have potent antimycobacterial activity in vitro at concentrations below 1 μg/ml. Here, two selected compounds, 2,4-disubstituted pyridine derivatives 11 and 15, revealed significant bactericidal activity against Mycobacterium tuberculosis localized intracellularly within human macrophages, as well as against biofilm-forming tubercle bacilli. Mutants were selected that were resistant to the investigated compounds at an efficiency similar to that identified in the presence of the first line antituberculosis drug rifampicin. The resistant mutants were viable in the presence of the tested compounds exclusively on solid media. Genome-wide sequencing of the mutants selected in the presence of compound 11 revealed the accumulation of nonsynonymous mutations in the mmpR5 gene encoding a transcriptional repressor of the MmpS5-MmpL5 efflux pump, whose upregulation has been associated with bedaquiline resistance. The depletion of MmpR5 in wild-type M. tuberculosis using CRISPR–Cas9 technology increased the resistance of this strain to compound 11. Mass spectrometry-based proteomics (LC–MS/MS) of wild-type tubercle bacilli growing in subinhibitory concentrations of compounds 11 or 15 revealed 15 overproduced proteins not detectable in the control cells, including virulence-related proteins.

Multivariate mapping is a technique in which multivariate data are encoded into a single map. A variety of design solutions for multivariate mapping refers to the number of phenomena mapped, the map type, and the visual variables applied. Unlike other authors who have mainly evaluated bivariate maps, in our empirical study we compared three solutions when mapping four variables: two types of multivariate maps (intrinsic and extrinsic) and a simple univariate alternative (serving as a baseline). We analysed usability performance metrics (answer time, answer accuracy, subjective rating of task difficulty) and eye-tracking data. The results suggested that experts used all the tested maps with similar results for answer time and accuracy, even when using four-variable intrinsic maps, which is considered to be a challenging solution. However, eye-tracking data provided more nuances in relation to the difference in cognitive effort evoked by the tested maps across task types.

The authors attempt to visualise nature conservation issues in the Podlaskie Voivodship using a series of small-scale thematic maps. Publicly available spatial and statistical data was used. The authors discuss the effects of applying the adopted methodological solutions. Some of them present a new approach to cartographic visualisation of spatial data. It may be an opportunity to look at various visualisation methods, their effectiveness, and the possibility of visualising nature issues on small-scale thematic maps.

Mycobacteria Prim-PolC performs short gap synthesis following removal of lesions during excision repair. Here the authors resolve crystal structures of pre- and post-catalytic Prim-PolC complexes bound to gapped DNA substrates to define its mechanism.

Chitosan is a very promising material for tissue model printing. It is also known that the introduction of chemical modifications to the structure of the material in the form of methacrylate groups makes it very attractive for application in the bioprinting of tissue models. The aim of this work is to study the characteristics of biomaterials containing chitosan (BCH) and its methacrylated equivalent (BCM) in order to identify differences in their usefulness in 3D bioprinting technology. It has been shown that the BCM material containing methacrylic chitosan is three times more viscous than its non-methacrylated BCH counterpart. Additionally, the BCM material is characterized by stability in a larger range of stresses, as well as better printability, resolution, and fiber stability. The BCM material has higher mechanical parameters, both mechanical strength and Young's modulus, than the BCH material. Both materials are ideal for bioprinting, but BCM has unique rheological properties and significant mechanical resistance. In addition, biological tests have shown that the addition of chitosan to biomaterials increases cell proliferation, particularly in 3D-printed models. Moreover, modification in the form of methacrylation encourages reduced toxicity of the biomaterial in 3D constructs. Our investigation demonstrates the suitability of a chitosan-enhanced biomaterial, specifically methacrylate-treated, for application in tissue engineering, and particularly for tissues requiring resistance to high stress, i.e., vascular or cartilage models. [ABSTRACT FROM AUTHOR]

The aim of this study was to assess the relationship of different chronic diseases with immunonutritional markers in the senior population. Methods: this study included 1190 hospitalized geriatric patients. The criteria to participate were ability to communicate, given consent and C-reactive protein (CRP) lower than 6 mg/dL. Results: the mean age of the study population was 81.7 ± 7.6 years. NLR (neutrophil-to-lymphocyte ratio), LMR (lymphocyte-to-monocyte ratio), MWR (monocyte-to-white blood cell ratio), SII (systemic immune–inflammation index), PNI (prognostic nutritional index) and CAR (C-reactive protein-to-albumin ratio) were related to age. NLR and MWR were higher, while LMR, PLR (platelet-to-lymphocyte ratio and SII were lower in men. All markers were related to BMI. NLR, LMR, LCR (lymphocyte-to-CRP ratio), MWR, PNI and CAR were related to several concomitant chronic diseases. In multivariate analyses, age and BMI were selected as independent predictors of all studied immunonutritional markers. Atrial fibrillation, diabetes mellitus and dementia appear most often in the models. PNI presented the most consistent statistical association with age, BMI and concomitant chronic diseases. Conclusions: this study reveals the pivotal role of aging and BMI in inflammatory marker levels and the association of immunonutritional markers with different chronic diseases. Atrial fibrillation seems to have the most dominant connection to the immunonutritional markers. [ABSTRACT FROM AUTHOR]

As a regional product, Koryciński cheese is one of the most important cheeses in the Podlasie region of Poland. In this study, the influence of technological processes, such as ripening, on shaping the organoleptic characteristics of cheese was determined. Korycin-type cheeses are produced from cow’s milk according to traditional technology. The ripening process lasted 466 h at 5 °C, 10 °C, and 15 °C. The aging temperature had a decisive influence on the number of esters and organic acids formed, which were analyzed by gas chromatography–mass spectrometry (GC/MS). The organoleptic properties of the cheeses were also related to the ripening temperature.

Nowadays a lot of people are trying to make maps, and especially digital maps. A wide range of computer tools and high graphic capabilities have together made maps increasingly popular and seemingly easy to prepare for any person who can use a computer. It seems necessary to verify the bases of the cartographic presentation methods. There is a need for a new, formalized view of the method as a sequence of steps from data collection, to correct presentation, to map. Two terms related to cartographic presentation should be distinguished in this article: “methods” and “forms.” A method is understood as the process by which data is transformed into a presentation. A form is understood as the end result of this process, i.e. the resulting graphical image or map. In the article five types of cartographic presentation are indicated. In the successive types, one can observe an increasing degree of complexity of cartographic presentation.

This article discusses the feminist implications of Louisa May Alcott’s 1863 Gothic story “A Whisper in the Dark,” which not only expresses the anxieties that the author experienced in response to her upbringing and her social reality, but also provides an extensive critique of patriarchal culture. The essay explores the subversive nature of the story by presenting it as a dark double to Charlotte Brontë’s Jane Eyre as well as by showing how the author mocks nineteenth-century sentimentality throughout.

The purpose of this study was to produce a multifaceted presentation of a small region. It was realised based on two aims: the collection of spatial and statistical data, and the analysis of the data from the perspective of physical and socio-economic geography. Mrozy commune (Polish – gmina Mrozy) is located in the eastern part of the Masovian voivodeship, in the county of Minsk (Polish – powiat miński).

This work examines the effect of selected process parameters on the diameter of uniform and heterogeneous fibers (with and without bead-on-string structures) and the size of beads obtained during the electrospinning process. A 23 factorial design was performed to determine the influence of the following factors: electrical voltage, flow rate and dynamic viscosity of the poly(vinylpyrrolidone) ethanolic solution. Factorial design enables the analysis of the mathematical relationship between the chosen factors and the response with a minimum number of experiments. The factor having the most significant impact on the size of beaded fibers and beads was the solution viscosity, while the voltage had the greatest influence on the bead-free fiber diameter. The interactions between the studied factors were also analyzed. It was found that the presented method can be used for the design of an optimal and cost-effective electrospinning process, allowing the desired product to be obtained with expected features.

The discovery in the cartographic collections of the Faculty of Geography and Regional Studies at the University of Warsaw of an original map by Charles Dupin – the first choropleth map – provided an opportunity to conduct a closer methodological analysis of the map and to investigate the subsequent development of this presentation method during the first half of 19th century. From relatively early on, the accepted principle was for choropleth map presentations to use statistical data still imprecisely referred to as relative, as well as using a distribution series as a method of generalizing data.

Ligase D is a conserved DNA repair protein complex that repairs double-strand breaks in stationary phase prokaryotes. Here the authors show that orthologous Ligase C has a role in base excision repair during stationary phase.

The authors of the article pay their attention to the lack of a generally accepted classification of the cartographic presentation methods. The classification, which was described in the Ratajski’s handbook (1989) in the mostly extensive way, is commonly used in the Polish literature. According to the authors, it would be appropriate to modify one of the types of symbols (quantitative ones) as an independent method of data presentation at the quantitative level, in addition to the method of diagram, choropleth, dot method and isoline one.

The masculine and feminine elements in Aleksandr Sokurov’s film Alexandra The aim of this article is to present the fundamentals of the Great Mother archetype in Aleksandr Sokurov’s film Alexandra using the anthropological-morphological method of analysis. The effect of meeting between the protagonists, Alexandra and Denis, and their interweaving, is a new person – a perfect being where the masculine and feminine is brought to a perfect harmony. Pierwiastki męskie i kobiece w filmie Aleksandra w reżyserii Aleksandra Sokurowa Artykuł jest poświęcony przedstawieniu rudymentów archetypu Wielkiej Matki w filmie Aleksandra w reżyserii Aleksandra Sokurowa za pomocą metody analizy antropologiczno-morfologicznej dzieła filmowego. Efektem spotkania bohaterów i ich „czynności tkackich” jest nowa osoba – istota idealna, w której równowaga pierwiastków męskich i kobiecych osiągnęła stan pełnej harmonii.

Thoughtful consideration of the enumeration unit size in choropleth map design is important to ensure the correct communication of spatial information. However, the enumeration unit size and its influence on pattern conveying in choropleth maps have not yet been the subject of in-depth empirical studies. This research aims to address this gap. We focused on the issue concerning whether the ability to recognize spatial patterns on an Equal Area Unit Map is related to the hexagonal enumeration unit size, defined by the number of pixels. The aim is to indicate the range of the enumeration unit sizes, namely, at what point the upper and lower borders of the range where the spatial patterns start, and where the end is visible and recognizable by users. To address this problem, we conducted an empirical study with 488 users. The results show that the enumeration unit size has an impact on the users’ spatial pattern recognition abilities. Choropleth maps with enumeration unit sizes of 26, 52, and 104 pixels were, in the majority, indicated by participants as those most suitable for indicating spatial patterns. This was in contrast to choropleth maps with enumeration unit sizes of 1664 and 3328 pixels, which users indicated as not being useful. However, there were some exceptions to this general finding. Thus, determining the optimal enumeration unit size is a challenging task, and requires further insightful investigations.