Search

The influence of human gut microbiota on health and disease is now commonly appreciated. Therefore, it is not surprising that microbiome research has found interest in the sports community, hoping to improve health and optimize performance. Comparative studies found new species or pathways that were more enriched in elites than sedentary controls. In addition, sport-specific and performance-level-specific microbiome features have been identified. However, the results remain inconclusive and indicate the need for further assessment. In this case-control study, we tested two athletic populations (i.e. strength athletes, endurance athletes) and a non-athletic, but physically active, control group across two acute exercise bouts, separated by a 2-week period, that measured explosive and high intensity fitness level (repeated 30-s all-out Wingate test (WT)) and cardiorespiratory fitness level (Bruce Treadmill Test). While we did not identify any group differences in alpha and beta diversity or significant differential abundance of microbiome components at baseline, one-third of the species identified were unique to each group. Longitudinal sample (pre- and post-exercise) analysis revealed an abundance of Alistipes communis in the strength group during the WT and 88 species with notable between-group differences during the Bruce Test. SparCC recognized Bifidobacterium longum and Bifidobacterium adolescentis, short-chain fatty acid producers with probiotic properties, species strongly associated with VO2max. Ultimately, we identified several taxa with different baseline abundances and longitudinal changes when comparing individuals based on their VO2max, average power, and maximal power parameters. Our results confirmed that the health status of individuals are consistent with assumptions about microbiome health. Furthermore, our findings indicate that microbiome features are associated with better performance previously identified in elite athletes.

Energy from biogas is widely available, inexpensive, and often contributes to waste management, making it one of the most promising renewable energy sources. The main factors influencing this process’ efficiency include the substrates’ chemical composition, temperature, and digester load. This paper presents the possibilities offered by a biogas plant built at a farm specialising in dairy cows. The dependence of the power generated in the micro biogas plant on its technical parameters was analysed in detail. Studies carried out by the authors in an agricultural microgas plant (with an electrical output of 40 kW) have shown that they are designed to maintain continuous energy production, despite changing process parameters such as digester mass level, biogas height, temperature or slurry flow into the digester. However, from the point of view of the amount of electricity generated, changes would have to be made to the design of the biogas plant. Firstly, a more powerful generator would have to be installed to cover the electricity requirements of the equipment installed in the biogas plant so that power close to the rated capacity of the biogas plant is still sent to the grid. Secondly, replacing the two existing agitators of the digestion mass (9 kW each) with more agitators of lower power (e.g., four agitators of 4.5 kW each) would be necessary. These should be programmed so that one of the agitators operates at any given time (the operating time of a given agitator should depend on the composition of the digestate).

Abstract Numeric sex chromosome abnormalities are commonly associated with an increased cancer risk. Here, we report a 14-year-old boy with a rare mosaic 45, X/48, XYYY karyotype presenting with subtle dysmorphic features and relative height deficiency, requiring growth hormone therapy. As only 12 postnatal cases have been described so far with very limited follow-up data, to assess the proband’s long-term prognosis, including cancer risk, we performed high-throughput single-cell RNA sequencing (scRNA-seq) analysis. Although comprehensive cytogenetic analysis showed seemingly near perfect balance between 45, X and 48, XYYY cell populations, scRNA-seq revealed widespread differences in genotype distribution among immune cell fractions, specifically in monocytes, B- and T-cells. These results were confirmed at DNA level by digital-droplet PCR on flow-sorted immune cell types. Furthermore, deregulation of predominantly autosomal genes was observed, including TCL1A overexpression in 45, X B-lymphocytes and other known genes associated with hematological malignancies. Together with the standard hematological results, showing increased fractions of monocytes and CD4+/CD8+T lymphocytes ratio, long-term personalized hemato-oncological surveillance was recommended in the reported patient.

IntroductionPolycythemia vera (PV) and essential thrombocythemia (ET) are diseases driven by canonical mutations in JAK2, CALR, or MPL gene. Previous studies revealed that in addition to driver mutations, patients with PV and ET can harbor other mutations in various genes, with no established impact on disease phenotype. We hypothesized that the molecular profile of patients with PV and ET is dynamic throughout the disease.MethodsIn this study, we performed a 37-gene targeted next-generation sequencing panel on the DNA samples collected from 49 study participants in two-time points, separated by 78-141 months. We identified 78 variants across 37 analyzed genes in the study population.ResultsBy analyzing the change in variant allele frequencies and revealing the acquisition of new mutations during the disease, we confirmed the dynamic nature of the molecular profile of patients with PV and ET. We found connections between specific variants with the development of secondary myelofibrosis, thrombotic events, and response to treatment. We confronted our results with existing conventional and mutation-enhanced prognostic systems, showing the limited utility of available prognostic tools.DiscussionThe results of this study underline the significance of repeated molecular testing in patients with PV and ET and indicate the need for further research within this field to better understand the disease and improve available prognostic tools.

Abstract Background Bilateral parotid gland tumors account for up to 3% of cases. In this group, the vast majority are Warthin’s tumors. However, bilateral presentations of other parotid gland tumor entities is also possible, an example of which is a basal cell adenoma (BCA). Bilateral BCA is extremely rare, which could cause misdiagnosing it as a Warthin tumor. Case presentation The current study reports the unique case of a 48-year-old woman who presented with a 6-month history of slowly growing masses located bilaterally in the parotid region, surgically treated with 5-year follow-up (no recurrence, normal facial nerve function). Magnetic resonance imaging (MRI) revealed three lesions: two in the superficial and deep lobes of the right parotid gland, and one in the superficial lobe of the left parotid gland. A total parotidectomy with facial nerve preservation was performed on the right side, and superficial parotidectomy on the left side 6 months later. Histopathological examination confirmed that all three tumors were BCAs. Molecular analysis didn’t show any strong, potential of unknown clinical significance in the studied sample. Conclusions Multifocal bilateral lesions of the parotid gland are usually Warthin tumors. Detailed preoperative diagnostics including MRI and histopathological examination is essential to avoid misdiagnosing BCA and Warthin tumors. To our best knowledge, no case of synchronous bilateral multifocal basal cell adenomas of the parotid gland has been reported in English literature so far.

The personal archive of Adam Tarn (1902–1975), the first editor-in-chief of the Dialog monthly and a translator, critic, novelist, and playwright, includes notes with loose ideas for plays, notes to his unfinished book on Chekhov, and minor literary attempts, which he corrected, set aside and later expanded. This material, which Tarn’s heirs kindly offered me for study, constitutes a valuable proof the creative process and the evolution of the writing of the Dialog’s editor-in-chief. In the context of Obraz ojca w czterech ramach [Father’s Image in Four Frames] (1934), Tarn’s début novel and the only one he completed, the novel Kameleon [Chameleon] is an intriguing item in the author’s collection of unfinished works. In this article I shall discuss the material contained in Tarn’s personal archive. This study offers an insight into his “internal laboratory” of writing and enables one to read it from, e.g., the biographical perspective. The dating of the collected material can be only approximated. The earliest surviving prose attempt written in French probably came from the interwar period, while the final notes were composed during his émigré period after 1968, when Tarn was working on his unfinished book on Chekhov.

Abstract Background Pollen development in the anther in angiosperms depends on complicated cellular interactions associated with the expression of gametophytic and sporophytic genes which control fundamental processes during microsporo/gametogenesis, such as exo/endocytosis, intracellular transport, cell signaling, chromatin remodeling, and cell division. Most if not all of these cellular processes depend of local concentration of calcium ions (Ca2+). Work from our laboratory and others provide evidence that calreticulin (CRT), a prominent Ca2+-binding/buffering protein in the endoplasmic reticulum (ER) of eukaryotic cells, may be involved in pollen formation and function. Here, we show for the first time the expression pattern of the PhCRT1 gene and CRT accumulation in relation to exchangeable Ca2+ in Petunia hybrida developing anther, and discuss probable roles for this protein in the male gametophyte development. Results Using northern hybridization, western blot analysis, fluorescent in situ hybridization (FISH), immunocytochemistry, and potassium antimonate precipitation, we report that PhCRT1 is highly expressed in the anther and localization pattern of the CRT protein correlates with loosely bound (exchangeable) Ca2+ during the successive stages of microsporo/gametogenesis. We confirmed a permanent presence of both CRT and exchangeable Ca2+ in the germ line and tapetal cells, where these factors preferentially localized to the ER which is known to be the most effective intracellular Ca2+ store in eukaryotic cells. In addition, our immunoblots revealed a gradual increase in CRT level from the microsporocyte stage through the meiosis and the highest CRT level at the microspore stage, when both microspores and tapetal cells show extremely high secretory activity correlated with the biogenesis of the sporoderm. Conclusion Our present data provide support for a key role of CRT in developing anther of angiosperms – regulation of Ca2+ homeostasis during pollen grains formation. This Ca2+-buffering chaperone seems to be essential for pollen development and maturation since a high rate of protein synthesis and protein folding within the ER as well as intracellular Ca2+ homeostasis are strictly required during the multi-step process of pollen development.

Introduction: Brain metastases (BM) severely affect the prognosis and quality of life of patients with NSCLC. Recently, molecularly targeted agents were found to have promising activity against BM in patients with NSCLC whose primary tumors carry “druggable” mutations. Nevertheless, it remains critical to identify specific pathogenic alterations that drive NSCLC-BM and that can provide novel and more effective therapeutic targets. Methods: To identify potentially targetable pathogenic alterations in NSCLC-BM, we profiled somatic copy number alterations (SCNAs) in 51 matched pairs of primary NSCLC and BM samples from 33 patients with lung adenocarcinoma and 18 patients with lung squamous cell carcinoma. In addition, we performed multiregion copy number profiling on 15 BM samples and whole-exome sequencing on 40 of 51 NSCLC-BM pairs. Results: BM consistently had a higher burden of SCNAs compared with the matched primary tumors, and SCNAs were typically homogeneously distributed within BM, suggesting BM do not undergo extensive evolution once formed. By comparing focal SCNAs in matched NSCLC-BM pairs, we identified putative BM-driving alterations affecting multiple cancer genes, including several potentially targetable alterations in genes such as CDK12, DDR2, ERBB2, and NTRK1, which we validated in an independent cohort of 84 BM samples. Finally, we identified putative pathogenic alterations in multiple cancer genes, including genes involved in epigenome editing and 3D genome organization, such as EP300, CTCF, and STAG2, which we validated by targeted sequencing of an independent cohort of 115 BM samples. Conclusions: Our study represents the most comprehensive genomic characterization of NSCLC-BM available to date, paving the way to functional studies aimed at assessing the potential of the identified pathogenic alterations as clinical biomarkers and targets.

Familial hypercholesterolemia (FH) is an autosomal-dominant disorder caused mainly by substitutions in the low-density lipoprotein receptor (LDLR) gene, leading to an increased risk of premature cardiovascular diseases. Tremendous advances in sequencing techniques have resulted in the discovery of more than 3000 variants of the LDLR gene, but not all of them are clinically relevant. Therefore, functional studies of selected variants are needed for their proper classification. Here, a single-cell, kinetic, fluorescent LDL uptake assay was applied for the functional analysis of LDLR variants in a model of an LDLR-deficient human cell line. An LDLR-defective HEK293T cell line was established via a CRISPR/Cas9-mediated luciferase–puromycin knock-in. The expressing vector with the LDLR gene under the control of the regulated promoter and with a reporter gene has been designed to overproduce LDLR variants in the host cell. Moreover, an LDLR promoter–luciferase knock-in reporter system has been created in the human cell line to study transcriptional regulation of the LDLR gene, which can serve as a simple tool for screening and testing new HMG CoA reductase-inhibiting drugs for atherosclerosis therapy. The data presented here demonstrate that the obtained LDLR-deficient human cell line HEK293T-ldlrG1 and the dedicated pTetRedLDLRwt expression vector are valuable tools for studying LDL internalization and functional analysis of LDLR and its genetic variants. Using appropriate equipment, LDL uptake to a single cell can be measured in real time. Moreover, the luciferase gene knock-in downstream of the LDLR promotor allows the study of promoter regulation in response to diverse conditions or drugs. An analysis of four known LDLR variants previously classified as pathogenic and benign was performed to validate the LDLR-expressing system described herein with the dedicated LDLR-deficient human cell line, HEK293T-ldlrG1.

The soils that sustained damage from the mining industry are threatened with high salinity. The aim of the research involved assessing the impact of drilling wastes on the salinity of soils, and the influence of salinity on the germination and growth of various grass species. The research involved the energy, germination capacity and growth of four grass species: tall fescue Festuca arundinacea (cv. Odys), red fescue Festuca rubra (cv. Areta), perennial ryegrass Lolium perenne (cv. Gazon) and smooth meadow grass Poa pratensis (cv. Alicja) in the soils with various amount of drilling wastes addition and different salinity. The drilling waste addition in the amount of 5%, 10%, 15%, 20%, 30% (v/v) (pH=4.1, EC=8.84 µS/cm) significantly increased the salinity of the prepared mixtures to the levels of >2.5 dS/m, determined as harmful for most plants. Studies indicated that 5%, 10% and 15% (v/v) drill cuttings addition does not inhibit the growth of the considered grasses, while at the 25% addition of drill cuttings, the length of seedlings and roots is halved in comparison to the control sample without drilling waste addition. The mixture with 30% drilling waste addition, characterized by the salinity of 18 dS/m, inhibits the growth of all considered plant species. The conducted discrimination analysis indicated that cv. Gazon and cv. Odys differ from the other considered grass species, exhibiting the highest resistance to salinity caused by drilling waste addition. In turn, cv. Alicja was characterized by the lowest tolerance to salinity.

The article briefly confronts different research perspectives on epistolography. The fundamental Teoria listu (Theory of a Letter) by Stefania Skwarczyńska is indicated as an important reference for many concepts. The category of a beautiful letter proposed by Skwarczyńska is presented by the example of the correspondence between Wisława Szymborska and Kornel Filipowicz — Listy. Najlepiej w życiu ma twój kot (Letters. Your Cat Has the Best in Life). The letters of the future Nobel Prize winner and the master of the storyteller harmoniously combine life and literary elements. They are characterised by irony and humour, which are specific codes of communication. Their form and concepts are evidence of the correspondents’ passionate and strong relationship. The sensitivity to nature shown in them was presented in the context of empathetic imagination. It was linked to both Filipowicz’s biographical experiences and his creative practices, threads that help in the origin of texts such as the short story Krajobraz doskonały (Perfect Landscape) from the volume Śmierć mojego antagonisty (Death of My Antagonist). Nowadays, the need for interdisciplinarity has been pointed out as one of the most important postulates concerning the study of epistolography. The bilateral nature of biography and correspondence continues to be an inspiring issue. The communication paradox of the letter, which is both a dialogue and a monologue, also returns in the investigations of researchers. In the context of the aforementioned studies, the letter continues to fascinate as a phenomenon on the borderline of life and art. For some artists, the letter had a community- creating power. This very meaning was especially emphasised in the perspective of searching for spiritual community, tightening important bonds, lasting and deep friendships based on certain rules. Volumes of correspondence of writers-friends, among others, Julian Tuwim, were cited as an example. The article is an invitation to develop and expand the study of both published correspondence, as well as letters resting in archives in Poland and abroad.

Crop fertilization with sulfur is an important part of agricultural practices, as is the systematic increase in soil organic matter content. Materials of waste origin constitute a source of plant-available sulfur, as well as soil organic matter. The study was to verify the hypothesis assuming that combining waste sulfur pulp and its mixtures with organic materials enables simultaneous soil enrichment with readily available sulfur and organic matter. A 240-day incubation experiment was conducted, on two soils: very light and heavy; with two sulfur doses applied to each soil (20 and 40 mg S/kg d.m. for very light soil, and 30 and 60 mg S/kg d.m. for heavy soil). The sulfate sulfur content in the incubated soil material, treated with the addition of sulfur pulp and its mixtures with organic materials, increased significantly up to day 60 and then decreased. The application of these materials significantly increased the content of available sulfur and decreased the pH value of the incubated material. The effect of the introduced materials on dehydrogenase activity depended on soil granulometric composition (the impact of the applied materials on the activity of these enzymes in very light soil was small, and in heavy soil, their activity was usually limited by the presence of introduced materials). Application of the studied materials had little effect on the total organic carbon content in the incubated soil material (a significant change in the value of this parameter, in relation to the control soil, was recorded in some treatments of heavy soil).

Background: It has been demonstrated that Egfl7 promotes tumor cell escape from immunity by downregulating the activation of tumor blood vessels. Aim: to analyze mRNA expression of EGFL7 within the tumor microenvironment of high-grade ovarian serous carcinoma and its association with a number of intraepithelial CD4+/CD8+ lymphocytes and ICAM-1 expression. Methods: qPCR analysis of EGFL7 mRNA in cancer cells and adjacent stromal endothelium microdissected from formalin-fixed paraffin-embedded tumors of 59 high-grade ovarian serous carcinoma patients, was performed. Infiltration of intraepithelial lymphocytes (CD4+/CD8+) and expression of ICAM-1 were evaluated by immunohistochemistry and compared between tumors with different statuses of EGFL7 expression. Results: EGFL7 was expressed in cancer cells (9/59, 15.25%), endothelium (8/59, 13.56%), or both cancer cells and adjacent endothelium (4/59, 6.78%). ICAM-1 was expressed on cancer cells (47/59, 79.66%), stromal endothelium (46/59, 77.97%), or both epithelium and endothelium (40 of 59, 67.8%). EGFL7-positivity of cancer cells and endothelium was associated with lower intraepithelial inflow of CD4+ (p = 0.022 and p = 0.029, respectively) and CD8+ lymphocytes (p = 0.004 and p = 0.031, respectively) but impact neither epithelial nor endothelial ICAM-1 expression (p = 0.098 and p = 0.119, respectively). The patients’ median follow-up was 23.83 months (range 1.07–78.07). Lack of prognostic significance of EGFL7-status and ICAM-1 expression was notified. Conclusion: EGFL7 is activated in the cancer cells as frequently as in the endothelium of human high-grade ovarian serous carcinoma. Activation of EGFL7 in cancer cells and/or endothelial cells could negatively impact diapedesis regardless of localization.

Pollen tube growth depends on several complex processes, including exo/endocytosis, cell wall biogenesis, intracellular transport, and cell signaling. Our previous results provided evidence that calreticulin (CRT)—a prominent calcium (Ca2+)-buffering molecular chaperone in the endoplasmic reticulum (ER) lumen—is involved in pollen tube formation and function. We previously cloned and characterized the CRT gene belonging to the CRT1/2 subgroup from Petunia hybrida (PhCRT1/2), and found that post-transcriptional silencing of PhCRT1/2 expression strongly impaired pollen tube growth in vitro. Here, we report cloning of a new PhCRT3a homolog; we identified the full-length cDNA sequence and described its molecular characteristics and phylogenetic relationships to other plant CRT3 genes. Using an RNA interference (RNAi) strategy, we found that knockdown of PhCRT3a gene expression caused numerous defects in the morphology and ultrastructure of cultivated pollen tubes, including disorganization of the actin cytoskeleton and loss of cytoplasmic zonation. Elongation of siPhCRT3a pollen tubes was disrupted, and some of them ruptured. Our present data provide the first evidence that PhCRT3a expression is required for normal pollen tube growth. Thus, we discuss relationships between diverse CRT isoforms in several interdependent processes driving the apical growth of the pollen tube, including actomyosin-dependent cytoplasmic streaming, organelle positioning, vesicle trafficking, and cell wall biogenesis.

Measuring various biochemical and cellular components in the blood is a routine procedure in clinical practice. Human serum contains hundreds of diverse proteins secreted from all cells and tissues in healthy and diseased states. Moreover, some serum proteins have specific strong interactions with other blood components, but most interactions are probably weak and transient. One of the serum proteins is butyrylcholinesterase (BChE), an enzyme existing mainly as a glycosylated soluble tetramer that plays an important role in the metabolism of many drugs. Our results suggest that BChE interacts with plasma proteins and forms much larger complexes than predicted from the molecular weight of the BChE tetramer. To investigate and isolate such complexes, we developed a two-step strategy to find specific protein–protein interactions by combining native size-exclusion chromatography (SEC) with affinity chromatography with the resin that specifically binds BChE. Second, to confirm protein complexes′ specificity, we fractionated blood serum proteins by density gradient ultracentrifugation followed by co-immunoprecipitation with anti-BChE monoclonal antibodies. The proteins coisolated in complexes with BChE were identified by mass spectroscopy. These binding studies revealed that BChE interacts with a number of proteins in the human serum. Some of these interactions seem to be more stable than transient. BChE copurification with ApoA-I and the density of some fractions containing BChE corresponding to high-density lipoprotein cholesterol (HDL) during ultracentrifugation suggest its interactions with HDL. Moreover, we observed lower BChE plasma activity in individuals with severely reduced HDL levels (≤20 mg/dL). The presented two-step methodology for determination of the BChE interactions can facilitate further analysis of such complexes, especially from the brain tissue, where BChE could be involved in the pathogenesis and progression of AD.

A copper film modified glassy carbon electrode (CuF/GCE) and a novel copper film with carbon nanotubes modified screen-printed electrode (CuF/CN/SPE) for anodic stripping voltammetric measurement of ultratrace levels of Cd(II) are presented. During the development of the research procedure, several main parameters were investigated and optimized. The optimal electroanalytical performance of the working electrodes was achieved in electrolyte 0.1 M HCl and 2 × 10−4 M Cu(II). The copper film modified glassy carbon electrode exhibited operation in the presence of dissolved oxygen with a calculated limit of detection of 1.7 × 10−10 M and 210 s accumulation time, repeatability with RSD of 4.2% (n = 5). In the case of copper film with carbon nanotubes modified screen-printed electrode limit of detection amounted 1.3 × 10−10 M for accumulation time of 210 s and with RSD of 4.5% (n = 5). The calibration curve has a linear range in the tested concentration of 5 × 10−10–5 × 10−7 M (r = 0.999) for CuF/GCE and 3 × 10−10–3 × 10−7 M (r = 0.999) for CuF/CN/SPE with 210 s accumulation time in both cases. The used electrodes enable trace determination of cadmium in different environmental water samples containing organic matrix. The validation of the proposed procedures was carried out through analysis certified reference materials: TM-25.5, SPS-SW1, and SPS-WW1.

The aim of the study was to compare conducting economic activity by people covered by social insurance andby social insurance for farmers in the years 2013-2017. The former included old-age pension insurance, disability andsurvivors’ insurance, sickness insurance, accident and health insurance as well as contributions for the Labour Fund. Socialinsurance for farmers was defined based on old-age and disability insurance, sickness insurance, accident insurance andmaternity insurance. Monthly contributions for social insurance for persons conducting non-agricultural economic activitywere three times higher in the public system of social insurance (ZUS, or National Social Insurance Institution). However,when health insurance and Labuor Fund contributions were included, they were five times higher than agricultural socialinsurance (KRUS, or Agricultural Social Insurance Fund). The share of social and health insurance in the income of personscommencing economic activity was more than twice higher than in the income of persons insured by KRUS.

Poland, due to the estimated shale gas deposits amounting to 346-768 billion m3 has become one of the most attractive regions for shale gas exploration in Europe. Throughout the period 2010-2015, 72 exploratory drillings have been made (as of 4.01.2016) while hydraulic fracturing was carried out 25 times. Employing new drilling and shale gas prospecting technologies raises a question pertaining to their impact on the environment. The number of chemical compounds used (approximately 2000) for the production of new technological fluids may potentially pollute the environment. The fact that the composition of these fluids remains undisclosed hinders the assessment of their impact on the environment and devising optimal methods for managing this type of waste. The presented work indicates the chemical compounds which may infiltrate to groundwater, identified on the basis of technological fluids characteristics, as well as the review of studies pertaining to their impact on potable water carried out in the United States. The study focused on marking heavy metals, calcium, sodium, magnesium, potassium, chlorides and sulphates in the surface waters collected in proximity of Lewino well.

Tropomyosin is a two-chain coiled coil protein, which together with the troponin complex controls interactions of actin with myosin in a Ca2+-dependent manner. In fast skeletal muscle, the contractile actin filaments are regulated by tropomyosin isoforms Tpm1.1 and Tpm2.2, which form homo- and heterodimers. Mutations in the TPM2 gene encoding isoform Tpm2.2 are linked to distal arthrogryposis and congenital myopathy—skeletal muscle diseases characterized by hyper- and hypocontractile phenotypes, respectively. In this work, in vitro functional assays were used to elucidate the molecular mechanisms of mutations Q93H and E97K in TPM2. Both mutations tended to decrease actin affinity of homo-and heterodimers in the absence and presence of troponin and Ca2+, although the effect of Q93H was stronger. Changes in susceptibility of tropomyosin to trypsin digestion suggested that the mutations diversified dynamics of tropomyosin homo- and heterodimers on the filament. The presence of Q93H in homo- and heterodimers strongly decreased activation of the actomyosin ATPase and reduced sensitivity of the thin filament to [Ca2+]. In contrast, the presence of E97K caused hyperactivation of the ATPase and increased sensitivity to [Ca2+]. In conclusion, the hypo- and hypercontractile phenotypes associated with mutations Q93H and E97K in Tpm2.2 are caused by defects in Ca2+-dependent regulation of actin–myosin interactions.

Alzheimer’s disease (AD), the most common form of dementia in elderly individuals, is marked by progressive neuron loss. Despite more than 100 years of research on AD, there is still no treatment to cure or prevent the disease. High levels of amyloid-β (Aβ) plaques and neurofibrillary tangles (NFTs) in the brain are neuropathological hallmarks of AD. However, based on postmortem analyses, up to 44% of individuals have been shown to have high Aβ deposits with no clinical signs, due to having a “cognitive reserve”. The biochemical mechanism explaining the prevention of cognitive impairment in the presence of Aβ plaques is still unknown. It seems that in addition to protein aggregation, neuroinflammatory changes associated with aging are present in AD brains that are correlated with a higher level of brain iron and oxidative stress. It has been shown that iron accumulates around amyloid plaques in AD mouse models and postmortem brain tissues of AD patients. Iron is required for essential brain functions, including oxidative metabolism, myelination, and neurotransmitter synthesis. However, an imbalance in brain iron homeostasis caused by aging underlies many neurodegenerative diseases. It has been proposed that high iron levels trigger an avalanche of events that push the progress of the disease, accelerating cognitive decline. Patients with increased amyloid plaques and iron are highly likely to develop dementia. Our observations indicate that the butyrylcholinesterase (BChE) level seems to be iron-dependent, and reports show that BChE produced by reactive astrocytes can make cognitive functions worse by accelerating the decay of acetylcholine in aging brains. Why, even when there is a genetic risk, do symptoms of the disease appear after many years? Here, we discuss the relationship between genetic factors, age-dependent iron tissue accumulation, and inflammation, focusing on AD.

Familial hypercholesterolemia (FH) is the most common monogenic autosomal dominant disorder. FH results in an increased cardiovascular mortality rate. However, cardiovascular risk control factors enable the avoidance of approximately 80% of strokes and cardiovascular diseases. Therefore, early detection and implementation of lipid-lowering treatment is essential. In the present study, 57 pediatric patients aged 9.57 ± 3.26 years with FH were enrolled in the study. Researchers checked the lipid profile and performed the ultrasound imaging including intima-media thickness (IMT) measurement and echo (e)-tracking in the study group. Patients were treated with a low-cholesterol diet solely or along with pharmacological treatment with statins. Subsequently, patients were monitored for 12 months. The positive results of dietary treatment were observed in 40 patients. The efficacy of 12 months of nutritional therapy along with pharmacological treatment was reported in 27 patients. We observed a significant decrease in the carotid beta index stiffness and an insignificant decrease in the IMT in the group of patients treated with statins. The obtained data show that statin therapy in children with FH allow for the reduction of the degree of atherosclerotic vessel changes.

Background: The monogenic defect in familial hypercholesterolemia (FH) is detected in ∼40% of cases. The majority of mutation-negative patients have a polygenic cause of high LDL-cholesterol (LDL-C). We sought to investigate whether the underlying monogenic or polygenic defect is associated with the response to rosuvastatin. Methods: FH Individuals were tested for mutations in LDLR and APOB genes. A previously established LDL-C-specific polygenic risk score (PRS) was used to examine the possibility of polygenic hypercholesterolemia in mutation-negative patients. All of the patients received rosuvastatin and they were followed for 8 ± 2 months. A propensity score analysis was performed to evaluate the variables associated with the response to treatment. Results: Monogenic subjects had higher mean (±SD) baseline LDL-C when compared to polygenic (7.6 ± 1.5 mmol/L vs. 6.2 ± 1.2 mmol/L; p < 0.001). Adjusted model showed a lower percentage of change in LDL-C after rosuvastatin treatment in monogenic patients vs. polygenic subjects (45.9% vs. 55.4%, p < 0.001). The probability of achieving LDL-C targets in monogenic FH was lower than in polygenic subjects (0.075 vs. 0.245, p = 0.004). Polygenic patients were more likely to achieve LDL-C goals, as compared to those monogenic (OR 3.28; 95% CI: 1.23–8.72). Conclusion: Our findings indicate an essentially higher responsiveness to rosuvastatin in FH patients with a polygenic cause, as compared to those carrying monogenic mutations.

The aim of this study was a comparison of aortic valve calcium score (AVCS) between patients with hypercholesterolemia and genetic diagnosis of familial hypercholesterolemia with low-density lipoprotein receptor gene mutation (LDLR-M group), versus patients with hypercholesterolemia without LDLR gene mutation (LDLR-WT group). A total of 72 LDLR-M patients and 50 LDLR-WT patients were enrolled in the study and underwent CT as a part of an assessment of coronary calcium scoring. AVCS was determined and compared between the two patient groups. AVCS was significantly higher in the LDLR-M group in comparison to the LDLR-WT group (13.8 ± 37.9 vs. 0.94 ± 3.1, p = 0.03). The Yates' chi-squared test for independence revealed that LDLR mutation and AVCS were significantly dependable (Chi^2 = 6.106, p = 0.013). The LDLR mutation was a strong predictor of a high AVCS (OR 7.83, 95% CI 2.08-29.50, p = 0.002) on multivariate regression analysis. Among the traditional risk factors, age (odds ratio 1.12, 95% CI 1.05-1.18, p

The paper presents the analysis of the human heartbeat value under the physical exercise and rest condition. The test group was divided into four groups regarding age and sex. Measurements were performed using a mobile application and a blood pressure monitor. The phone's accuracy with respect to the blood pressure gauge was measured and the statistical significance of differences between age groups, sex and activity was also computed. Measurement inaccuracy varies from 4 to 12%. The mobile application accurately reproduces the pulse during rest.

Mastocytosis is characterized by excessive proliferation and accumulation of mast cells in skin and/or other organs. Two forms of the disease, cutaneous and systemic mastocytosis, differ significantly in symptomatology and clinical course. KIT mutations play an important role in the pathogenesis of the disease. The presence of p.D816V KIT mutation was detected in the vast majority of adults with systemic mastocytosis. The role of KIT mutations in childhood-onset mastocytosis remains a matter of discussion. More recent studies have shown that cutaneous mastocytosis, which is the most common clinical manifestation of the disease in children, has a genetic background. In contrast to adults, different types of KIT mutations have been described in pediatric and familial mastocytosis. The understanding of the molecular mechanisms in mastocytosis enables targeted therapy using tyrosine kinase inhibitors.

Late-onset Alzheimer’s disease (AD) is clinically characterized by a progressive decline of memory and other cognitive functions leading to the loss of the ability to perform everyday activities. Only a few drugs have been approved to treat AD dementia over the past century since the first AD patient was diagnosed. Drugs increasing the availability of neurotransmitters at synapses in the brain are used clinically in the treatment of AD dementia, and cholinesterase inhibitors (ChEIs) are the mainstay of the therapy. A detrimental effect on cognitive function has been reported in patients with pharmacological inhibition of acetylcholinesterase (AChE) by ChEIs and reduced butyrylcholinesterase (BChE) activity due to the single nucleotide polymorphisms. The BChE K-variant (rs1803274), the most common genetic variant of the BCHE gene, was thought to reduce enzyme activity reflecting the lower clinical response to rivastigmine in AD patients. During ChEIs therapy, patients carrying reduced-activity BChE do not present such improved attention like patients with the wild-type enzyme. On the other hand, alterations in the BCHE gene causing enzyme activity reduction may delay AD onset in patients at risk by preserving the level of cortical acetylcholine (ACh). Based on our previous results, we conclude that SNPs localized outside of the coding sequence, in 5’UTR (rs1126680) and/or intron 2 (rs55781031) of the BCHE gene, but not solely K-variant alteration (p.A539T) itself, are responsible for reduced enzyme activity. Therefore, we suspect that not BChE-K itself, but these coexisting SNPs (rs1126680 and rs55781031), could be associated with deleterious changes in cognitive decline in patients treated with ChEIs. Based on the results, we suggest that SNPs (rs1126680) and/or (rs55781031) genotyping should be performed to identify subjects at risk for lowered efficacy ChEIs therapy, and such patients should be treated with a lower rivastigmine dosage. Finally, our sequence analysis of the N-terminal end of N-BChE revealed evolutionarily conserved amino acid residues that can be involved in disulfide bond formation and anchoring of N-BChE in the cell membrane.

Butyrylcholinesterase (BChE) activity assay and inhibitor phenotyping can help to identify patients at risk of prolonged paralysis following the administration of neuromuscular blocking agents. The assay plays an important role in clinical chemistry as a good diagnostic marker for intoxication with pesticides and nerve agents. Furthermore, the assay is also commonly used for in vitro characterization of cholinesterases, their toxins and drugs. There is still lack of standardized procedure for measurement of BChE activity and many laboratories use different substrates at various concentrations. The purpose of this study was to validate the BChE activity assay to determine the best dilution of human serum and the most optimal concentration of substrates and inhibitors. Serum BChE activity was measured using modified Ellman's method applicable for a microplate reader. We present our experience and new insights into the protocol for high-throughput routine assays of human plasma cholinesterase activities adapted to a microplate reader. During our routine assays used for the determination of BChE activity, we have observed that serum dilution factor influences the results obtained. We show that a 400-fold dilution of serum and 5mM S-butyrylthiocholine iodide can be successfully used for the accurate measurement of BChE activity in human serum. We also discuss usage of various concentrations of dibucaine and fluoride in BChE phenotyping. This study indicates that some factors of such a multicomponent clinical material like serum can influence kinetic parameters of the BChE. The observed inhibitory effect is dependent on serum dilution factor used in the assay.