Microchip Isoelectric Focusing Applications

This review summarizes all available publications on isoelectric focusing carried out on planar microfabricated devices. Characteristic features of microchip isoelectric focusing (MC-IEF), such as resolution independence of separation length and compression of pH gradient, are discussed. An overview of materials used for microchip fabrication and developed detection strategies is given. Accomplishments in on-chip coupling of MC-IEF with other electrophoretic separation techniques or with mass spectrometry are briefly described. The review ends with alternative approaches to MC-IEF separation in terms of unusual sample introduction, kinds of support used for IEF separation, and in manner in which the pH gradient is generated

Clinoptilolite in Drinking Water Treatment for Ammonia Removal

In most countries today the removal of ammonium ions from drinking water has become almost a necessity. The natural zeolite clinoptiloliteis mined commercially in many parts of the world. It is a selective exchanger for the ammonium cation, and this has prompted its use in water treatment, wastewater treatment, swimming pools and fish farming. The work described in this paper provides dynamic data on cation exchange processes in clinoptilolite involving the NH4 +, Ca+2 and Mg+2 cations. We used material of natural origin – clinoptilolite from Nižný Hrabovec in Slovakia (particle-size 3–5 mm). The breakthrough capacity was determined by dynamic laboratory investigations, and we investigated the influence of thermal pretreatment of clinoptilolite and the concentration of regenerant solution (2, 5, and 10% NaCl). The concentrations of ammonium ion inputs in the tap water that we used were 10, 5, and 2 mg NH4 + l_1 and down to levels below 0.5 mg NH4 + l_1. The experimental results show that repeated pretreatment sufficiently improves the zeolite’s properties, and the structure of clinoptilolite remains unchanged during the loading and regeneration cycles. Ammonium removal capacities were increased by approximately 40 % and 20 % for heat-treated zeolite samples. There was no difference between the regenerates for 10% and 5% NaCl. We conclude that the use of zeolite is an attractive and promising method for ammonium removal

Lewatit S100 in Drinking Water Treatment for Ammonia Removal

Ammonium nitrogen is the most important form of nitrogen that can cause excessive algal growth and stimulate eutrophication in surface water. The purpose of this study is to investigate the possibility of removing ammonium from drinking water by means of an ion Exchange process. Polymeric Lewatit S100 material (particle-size 0.3–1.2 mm) was used. The breakthrough capacity was determined by dynamic laboratory investigations and the concentration of regenerant solution (5 and 10 % NaCl) was investigated. The concentration of ammonium ion inputs in the tap water that we used were 10, 5 and 2 mg NH4+ l_1 and down to levels below 0.5 mg NH4 + l_1. The experimental results show that the breakthrough capacity was very small at ammonium concentration 2 mg NH4 + l_1 compared to its breakthrough capacity at ammonium concentration 10 mg NH4 + l_1. There was no difference between regeneration by 10 and 5 % NaCl. We conclude that the use of Lewatit S100 is an attractive and promising method for ammonium concentration greater than 5 mg NH4 + l_1 and till 10 mg NH4 + l_1

DNA methylation profiles in a group of workers occupationally exposed to nanoparticles

The risk of exposure to nanoparticles (NPs) has rapidly increased during the last decade due to the vast use of nanomaterials (NMs) in many areas of human life. Despite this fact, human biomonitoring studies focused on the effect of NP exposure on DNA alterations are still rare. Furthermore, there are virtually no epigenetic data available. In this study, we investigated global and gene-specific DNA methylation profiles in a group of 20 long-term (mean 14.5 years) exposed, nanocomposite, research workers and in 20 controls. Both groups were sampled twice/day (pre-shift and post-shift) in September 2018. We applied Infinium Methylation Assay, using the Infinium MethylationEPIC BeadChips with more than 850,000 CpG loci, for identification of the DNA methylation pattern in the studied groups. Aerosol exposure monitoring, including two nanosized fractions, was also performed as proof of acute NP exposure. The obtained array data showed significant differences in methylation between the exposed and control groups related to long-term exposure, specifically 341 CpG loci were hypomethylated and 364 hypermethylated. The most significant CpG differences were mainly detected in genes involved in lipid metabolism, the immune system, lung functions, signaling pathways, cancer development and xenobiotic detoxification. In contrast, short-term acute NP exposure was not accompanied by DNA methylation changes. In summary, long-term (years) exposure to NP is associated with DNA epigenetic alterations

Neurodevelopmental and Epilepsy Phenotypes in Individuals With Missense Variants in the Voltage-Sensing and Pore Domains of KCNH5

Background and Objectives KCNH5 encodes the voltage-gated potassium channel EAG2/Kv10.2. We aimed to delineate the neurodevelopmental and epilepsy phenotypic spectrum associated with de novo KCNH5 variants.Methods We screened 893 individuals with developmental and epileptic encephalopathies for KCNH5 variants using targeted or exome sequencing. Additional individuals with KCNH5 variants were identified through an international collaboration. Clinical history, EEG, and imaging data were analyzed; seizure types and epilepsy syndromes were classified. We included 3 previously published individuals including additional phenotypic details.Results We report a cohort of 17 patients, including 9 with a recurrent de novo missense variant p.Arg327His, 4 with a recurrent missense variant p.Arg333His, and 4 additional novel missense variants. All variants were located in or near the functionally critical voltage-sensing or pore domains, absent in the general population, and classified as pathogenic or likely pathogenic using the American College of Medical Genetics and Genomics criteria. All individuals presented with epilepsy with a median seizure onset at 6 months. They had a wide range of seizure types, including focal and generalized seizures. Cognitive outcomes ranged from normal intellect to profound impairment. Individuals with the recurrent p.Arg333His variant had a self-limited drug-responsive focal or generalized epilepsy and normal intellect, whereas the recurrent p.Arg327His variant was associated with infantile-onset DEE. Two individuals with variants in the pore domain were more severely affected, with a neonatal-onset movement disorder, early-infantile DEE, profound disability, and childhood death.Discussion We describe a cohort of 17 individuals with pathogenic or likely pathogenic missense variants in the voltage-sensing and pore domains of Kv10.2, including 14 previously unreported individuals. We present evidence for a putative emerging genotype-phenotype correlation with a spectrum of epilepsy and cognitive outcomes. Overall, we expand the role of EAG proteins in human disease and establish KCNH5 as implicated in a spectrum of neurodevelopmental disorders and epilepsy.</p

Ultra-Rare Genetic Variation in the Epilepsies : A Whole-Exome Sequencing Study of 17,606 Individuals

Sequencing-based studies have identified novel risk genes associated with severe epilepsies and revealed an excess of rare deleterious variation in less-severe forms of epilepsy. To identify the shared and distinct ultra-rare genetic risk factors for different types of epilepsies, we performed a whole-exome sequencing (WES) analysis of 9,170 epilepsy-affected individuals and 8,436 controls of European ancestry. We focused on three phenotypic groups: severe developmental and epileptic encephalopathies (DEEs), genetic generalized epilepsy (GGE), and non-acquired focal epilepsy (NAFE). We observed that compared to controls, individuals with any type of epilepsy carried an excess of ultra-rare, deleterious variants in constrained genes and in genes previously associated with epilepsy; we saw the strongest enrichment in individuals with DEEs and the least strong in individuals with NAFE. Moreover, we found that inhibitory GABA(A) receptor genes were enriched for missense variants across all three classes of epilepsy, whereas no enrichment was seen in excitatory receptor genes. The larger gene groups for the GABAergic pathway or cation channels also showed a significant mutational burden in DEEs and GGE. Although no single gene surpassed exome-wide significance among individuals with GGE or NAFE, highly constrained genes and genes encoding ion channels were among the lead associations; such genes included CACNAIG, EEF1A2, and GABRG2 for GGE and LGI1, TRIM3, and GABRG2 for NAFE. Our study, the largest epilepsy WES study to date, confirms a convergence in the genetics of severe and less-severe epilepsies associated with ultra-rare coding variation, and it highlights a ubiquitous role for GABAergic inhibition in epilepsy etiology.Peer reviewe

The Human Phenotype Ontology in 2024: phenotypes around the world.

The Human Phenotype Ontology (HPO) is a widely used resource that comprehensively organizes and defines the phenotypic features of human disease, enabling computational inference and supporting genomic and phenotypic analyses through semantic similarity and machine learning algorithms. The HPO has widespread applications in clinical diagnostics and translational research, including genomic diagnostics, gene-disease discovery, and cohort analytics. In recent years, groups around the world have developed translations of the HPO from English to other languages, and the HPO browser has been internationalized, allowing users to view HPO term labels and in many cases synonyms and definitions in ten languages in addition to English. Since our last report, a total of 2239 new HPO terms and 49235 new HPO annotations were developed, many in collaboration with external groups in the fields of psychiatry, arthrogryposis, immunology and cardiology. The Medical Action Ontology (MAxO) is a new effort to model treatments and other measures taken for clinical management. Finally, the HPO consortium is contributing to efforts to integrate the HPO and the GA4GH Phenopacket Schema into electronic health records (EHRs) with the goal of more standardized and computable integration of rare disease data in EHRs

Occupational asthma follow-up — which markers are elevated in exhaled breath condensate and plasma?

Objectives: To search for optimal markers in the exhaled breath condensate (EBC), plasma and urine that would reflect the activity/ severity of occupational asthma (OA) after the withdrawal from the exposure to the allergen. Material and Methods: Markers of oxidative stress: 8-iso-prostaglandin F2α (8-isoprostane, 8-ISO), malondialdehyde (MDA), 4-hydroxy-trans-2-nonenale (HNE), cysteinyl leukotrienes (LT) and LTB4 were determined using liquid chromatography and mass spectrometry in 43 subjects with immunological OA (49.3±11.8 years), removed from the exposure to the sensitizing agent 10.5±6.5 years ago; and in 20 healthy subjects (49.0±14.9 years). EBC was harvested both before and after the methacholine challenge test. In parallel, identical markers were collected in plasma and urine. The results were analyzed together with forced expiratory volume in one second (FEV1), blood eosinophils, immunoglobulin E (IgE) and eosinophilic cationic protein (ECP) and statistically evaluated (Spearman rank correlation rS, two- or one-sample t tests and alternatively Kruskal Wallis or pair Wilcoxon tests). Results: Several parameters of lung functions were lower in the patients (FEV1% predicted, MEF25% and MEF50%, Rtot%, p < 0.001). Shorter time interval since the removal from the allergen exposure correlated with higher ECP (rS = 0.375) and lower FEV1%, MEF25% and MEF50% after methacholine challenge (rS = -0.404, -0.425 and -0.532, respectively). In the patients, IgE (p < 0.001) and ECP (p = 0.009) was increased compared to controls. In EBC, 8-ISO and cysteinyl LTs were elevated in the asthmatics initially and after the challenge. Initial 8-ISO in plasma correlated negatively with FEV1 (rS = -0.409) and with methacholine PD20 (rS = -0.474). 8-ISO in plasma after the challenge correlated with IgE (rS = 0.396). Conclusions: The improvement in OA is very slow and objective impairments persist years after removal from the exposure. Cysteinyl LTs and 8-ISO in EBC and 8-ISO in plasma might enrich the spectrum of useful objective tests for the follow-up of OA

Clinoptilolite in Drinking Water Treatment for Ammonia Removal

In most countries today the removal of ammonium ions from drinking water has become almost a necessity. The natural zeolite clinoptiloliteis mined commercially in many parts of the world. It is a selective exchanger for the ammonium cation, and this has prompted its use in water treatment, wastewater treatment, swimming pools and fish farming. The work described in this paper provides dynamic data on cation exchange processes in clinoptilolite involving the NH4 +, Ca+2 and Mg+2 cations. We used material of natural origin – clinoptilolite from Nižný Hrabovec in Slovakia (particle-size 3–5 mm). The breakthrough capacity was determined by dynamic laboratory investigations, and we investigated the influence of thermal pretreatment of clinoptilolite and the concentration of regenerant solution (2, 5, and 10% NaCl). The concentrations of ammonium ion inputs in the tap water that we used were 10, 5, and 2 mg NH4 + l_1 and down to levels below 0.5 mg NH4 + l_1. The experimental results show that repeated pretreatment sufficiently improves the zeolite’s properties, and the structure of clinoptilolite remains unchanged during the loading and regeneration cycles. Ammonium removal capacities were increased by approximately 40 % and 20 % for heat-treated zeolite samples. There was no difference between the regenerates for 10% and 5% NaCl. We conclude that the use of zeolite is an attractive and promising method for ammonium removal.</p