Bioinspired polyethersulfone-based hollow fiber membranes as the scaffolds in renal assist device for protein-bound toxins removal from blood

Dissertation for obtaining the Master degree in Membrane EngineeringErasmus Mundus Master in Membrane EngineeringUsing bioartificial kidney is the promising approach for removal of non-dializable, proteinbound uremic toxins, which are responsible for high mortality and morbidity in treating kidney failure related conditions. Additionaly, bioartificial kidney device could perform the physiological roles of the kidney such as metabolic replacement, endocrine function and immunomodulation. In the current work two commercial polyethersulfone-based membranes, Gambro HCO 1100 and Membrana MicroPES TF10 used in haemofiltration and plasma separation applications respectively were investigated. To provide adequate cytocompatibility of the membrane biomimetic, biomimetic double layer coating was developed. First, the membranes were coated with musselinspired synthetic polydopamine film, following with the coating of Collagen Type IV. Transport properties of the coated and native membranes were investigated. Increase in pure water permeability of the coated HCO 1100 membranes was observed. Membrane surface hydrophilization was assumed as the major factor responsible for the effect. Membrane permeabilities for bovine serum albumin and immunoglobulin G solutions were studied. Significant increase in protein rejection was observed for double coated HCO 1100 membranes with small or no effect of the double coated MicroPES TF10 membranes. Next, formation of confluent monolayers of the renal epithelial cells on the membrane scaffolds was studied. Cell seeding strategy was developed and two seeding conditions were tested. Specifically, the cells were allowed to adhere to the biomimetic membranes passively, and the negative pressure was applied to facilitate cell adhesion. After cultivation in semi-batch conditions the monolayer formation was examined. Confluent monolayers were observed for the conditions with passive cell adherence for the both membranes. Cell contacts formation and cell polarization were confirmed with the staining for ZO-1 protein. Applying the pressure to facilitate cell adhesion, on the contrary, resulted in the loss of cell ability to form functional monolayers.EM3E Master is an Education Programme supported by the European Commission, the European Membrane Society (EMS), the European Membrane House (EMH), and a large international network of industrial companies, research centres and universitie

Novel copper-containing membrane monooxygenases (CuMMOs) encoded by alkane-utilizing Betaproteobacteria.

Copper-containing membrane monooxygenases (CuMMOs) are encoded by xmoCAB(D) gene clusters and catalyze the oxidation of methane, ammonia, or some short-chain alkanes and alkenes. In a metagenome constructed from an oilsands tailings pond we detected an xmoCABD gene cluster with <59% derived protein sequence identity to genes from known bacteria. Stable isotope probing experiments combined with a specific xmoA qPCR assay demonstrated that the bacteria possessing these genes were incapable of methane assimilation, but did grow on ethane and propane. Single-cell amplified genomes (SAGs) from propane-enriched samples were screened with the specific PCR assay to identify bacteria possessing the target gene cluster. Multiple SAGs of Betaproteobacteria belonging to the genera Rhodoferax and Polaromonas possessed homologues of the metagenomic xmoCABD gene cluster. Unexpectedly, each of these two genera also possessed other xmoCABD paralogs, representing two additional lineages in phylogenetic analyses. Metabolic reconstructions from SAGs predicted that neither bacterium encoded enzymes with the potential to support catabolic methane or ammonia oxidation, but that both were capable of higher n-alkane degradation. The involvement of the encoded CuMMOs in alkane oxidation was further suggested by reverse transcription PCR analyses, which detected elevated transcription of the xmoA genes upon enrichment of water samples with propane as the sole energy source. Enrichments, isotope incorporation studies, genome reconstructions, and gene expression studies therefore all agreed that the unknown xmoCABD operons did not encode methane or ammonia monooxygenases, but rather n-alkane monooxygenases. This study broadens the known diversity of CuMMOs and identifies these enzymes in non-nitrifying Betaproteobacteria

Radically extended modified mastectomy of t4b-dn0-3m0 primary inflammatory breast cancer as a tool to minimize the risk of recurrence

Introduction. Breast cancer (BC) is an important public health problem. These are BC patients in young age, hereditary (BRCA-associated) BC patients, inflammatory BC patients, synchronous cancer patients etc. In case of a large size of tumor or diffuse form of BC we often need to use extended surgery techniques in order to perform it radically and minimize a risk of recurrence. The aim of the study to evaluate the effectiveness of surgery approaches in inflammatory BC patients. Materials and research methods. Тhe first stage of our research was to evaluate extended surgical treatment using techniques by Handelhail and Beck on the risk of local recurrence. The study included 39 patients with breast cancer T4b-dN0-3M0 who in the period from 2014 to 2019 received complex treatment at National Cancer Institute of Ukraine. Besides neo- and adjuvant chemotherapy, an extended radical mastectomy using a technique by Handelhail and Beck was performed in 19 patients. In 20 patients a traditional (Madden) modified radical mastectomy was done. Results. When studying the data of a group of 20 patients we received a very high percentage of recurrence. Recurrences were classified as locoregional if they occurred in the ipsilateral breast or the axillary or supraclavicular lymph nodes, and as distant metastasis, if they occurred at any other site. Radical extended modified mastectomy consisted in the removal of the mammary gland and adjacent skin along the perimeter with subcutaneous tissue at a distance of 2-3 cm outward from the macroscopic edges of the gland, subclavian-axillary-subscapularis lymphadenectomy. The resulting defect was closed using a technique by Handelhail and Beck. The use of extended surgery made it possible to improve recurrence-free survival

Changes in the vulnerability of clinical manifestations of thyrotoxic cardiomyopathy as indicators of remote results of surgical treatment in patients with toxic goiter

The aim of the work – improvement of the evaluation method of remote results of surgical treatment in patients with toxic goiter, taking into account the changes in the severity of clinical manifestations of thyrotoxic cardiomyopathy Materials and methods. The research was conducted in 150 patients operated for toxic goiter of moderate severity (48) and severe (102). The method of estimation of remote results of surgical treatment, taking into account the regression of signs of thyrotoxic cardiomyopathy and gradation and division into good, satisfactory, unsatisfactory and ineffective treatment, was developed. Results and discussion. A good remote outcome of surgical treatment of thyrotoxicosis was achieved in 26 (54.2 %) patients with moderate thyrotoxicosis and in 9 (8.8%) patients with severe thyrotoxicosis, satisfactory in 21 (43.7 %) and 36 (35, 3%) cases respectively and unsatisfactory – in 1 (2.0 %) and 33 (32.4 %). Ineffective surgical treatment was recognized in 24 (23.5 %) patients. Unsuccessful results and ineffective surgical treatment of toxic goiter were found in patients with severe heart rate rhythm disorders, structural and functional changes in the cardiac muscle and functional classes II and III. Conclusions. Long-term results of surgical treatment of patients with toxic goiter are determined by the severity of thyrotoxic cardiomyopathy. In cases of severe thyrotoxicosis, the question of the need for surgical treatment should be put in the development of severe heart rhythm disorders and structural and functional changes in the heart muscle

Seasonal dynamics of methanotrophic bacteria in a boreal oil sands end-pit lake

Copyright © 2022 American Society for Microbiology. All Rights Reserved.NSERC CRD (CRDPJ478071-14); King Abdullah Scholarship Program (KASP) - Ministry of Education, Kingdom of Saudi Arabia; NSERC CGS-M; NSERC CRD (CRDPJ476388–14).Peer ReviewedBase Mine Lake (BML) is the first full-scale demonstration end pit lake for the oil sands mining industry in Canada. We examined aerobic methanotrophic bacteria over all seasons for 5 years in this dimictic lake. Methanotrophs comprised up to 58% of all bacterial reads in 16S rRNA gene amplicon sequencing analyses (median 2.8%), and up to 2.7 × 104 cells mL−1 of water (median 0.5 × 103) based on qPCR of pmoA genes. Methanotrophic activity and populations in the lake water were highest during fall turnover and remained high through the winter ice-covered period into spring turnover. They declined during summer stratification, especially in the epilimnion. Three methanotroph genera (Methylobacter, Methylovulum, and Methyloparacoccus) cycled seasonally, based on both relative and absolute abundance measurements. Methylobacter and Methylovulum populations peaked in winter/spring, when methane oxidation activity was psychrophilic. Methyloparacoccus populations increased in the water column through summer and fall, when methane oxidation was mesophilic, and also predominated in the underlying tailings sediment. Other, less abundant genera grew primarily during summer, possibly due to distinct CH4/O2 microniches created during thermal stratification. These data are consistent with temporal and spatial niche differentiation based on temperature, CH4 and O2. This pit lake displays methane cycling and methanotroph population dynamics similar to natural boreal lakes

Systemic therapy of inflammatory breast cancer with type 2 diabetes mellitus – Prevention of high risk of radiation-induced progression of tumor tissue cancer

Introduction. Cancer mortality in diabetic patients has been reported to increase moderately compared to non-diabetic patients. The aim of the study aimed to assess the findings and identify radiotherapy's role in the comprehensive care of diabetic IBC patients with various hyperglycemia correction strategies. Methods. Patients with diabetes have shown a higher risk of radiation-induced cancer progression for tumor tissue, especially for inflammatory form. For 7 patients, to continue systematic chemotherapy with a scheme change (consecutive anthracyclines-taxanes, 2-week interval) and insulin with individual scheme for hyperglycemia correction on the basis of glycemic control – 1st group, the other (7 patients) – 2nd group, was given radiotherapy to the breast gland and lymphatic drainage ways. 45–50 Grey was prescribed for 25-28 fractions (per 1.8-2.0 Grey), 46–50 Grey in 23-25 fractions were used for zones of regional metastasis and for hyperglycemia correction metformin 2000 mg/day PO divided q8-12hr with meal on the basis of glycemic control. Results. Assessed were the number of patients who qualified for surgery and overall survival for 24 months. 2nd group showed a superior response following resistance to prior systemic treatment. Thus, 5 (71.41%) of the 7 patients exhibited a consistent response – complete or partial regression. There were only 2 individuals (28.61%) who responded to treatment among the patients who maintained chemotherapy. Conclusions. Breast cancer of the aggressive IBC variety requires multidisciplinary treatment from breast surgery, medical, and radiation oncology. Patients with diabetes appear to experience more side effects from radiation therapy than patients without the disease. Hyperglycemia, higher total RT doses, and radiosensitizers are a few techniques that can improve the impact of RT on local-regional management. Local-regional control rates for IBC are increasing with an individual patient strategy. Metformin also improves insulin resistance and has anticancer benefits

Microbial Community Dynamics in Base Mine Lake, the First End-Pit Lake in the Alberta Oil sands Industry

Oil extraction from bitumen in the Athabasca region of northeastern Alberta, comprises a large segment of the Canadian economy. However, the process of oil extraction from surface mined oil sands ores results in diverse environmental issues including disturbance of land areas and habitats for wildlife, as well as production of large volumes of fluid tailings containing many compounds of concern for the environment. Land reclamation strategies of most oilsands operations propose the construction of end-pit lakes (EPL) to contain and biodegrade tailings, eventually becoming integrated into local watersheds. We used 16S/18S rRNA gene amplicon and metagenome sequencing to monitor prokaryotic and eukaryotic communities in the first full-scale pilot EPL of the Canadian oilsands, Base Mine Lake (BML) Fig. 1, over 6 years from 2015-2021, and compared them with communities from two active tailings ponds, Mildred Lake Setting Basin (MLSB) and Southwest in-Pit (SWIP), as well as with communities from a more natural freshwater body, Beaver Creek Reservoir (BCR). Alpha diversity in BML is intermediate to diversities in active tailings ponds and a natural lake, although highly variable with time, depth, and season. Microbial communities in BML resemble neither freshwater lake nor active tailings communities, although a structural shift of either microbial community occurred every year, and 2021 was a year of the most profound effect. Moreover, metabolic functions in BML also shifted every year, with the most dramatic shift for photosynthesis-related genes. Temporal changes among protists identified in BML were characterized by enrichment of species designated as picophytoplankton: Cryptomonas, Mychonastes, Trebouxiophyceae, and Dinobryon, and among bacterial genera by enrichment of common freshwater lake species or bacterioplankton: Ca. Fonsibacter, Sporichthyaceae, Ca. Planktophila, Microbacteriaceae ML602J-51, Ilumatobacteraceae CL500-29 group. Network analysis identified a potential microbial consortium between newly enriched species of picophytoplankton and bacterioplankton. Heterotrophic bacteria with streamlined genomes may overcome auxotrophic limitations by scavenging metabolites and other compounds produced by phototrophs presumably via chemotaxis behaviour. We propose that the formation of the consortium might serve as a biomarker for reclamation process of an oil sands tailings pond

GAL08, an Uncultivated Group of Acidobacteria, Is a Dominant Bacterial Clade in a Neutral Hot Spring.

GAL08 are bacteria belonging to an uncultivated phylogenetic cluster within the phylum Acidobacteria. We detected a natural population of the GAL08 clade in sediment from a pH-neutral hot spring located in British Columbia, Canada. To shed light on the abundance and genomic potential of this clade, we collected and analyzed hot spring sediment samples over a temperature range of 24.2-79.8°C. Illumina sequencing of 16S rRNA gene amplicons and qPCR using a primer set developed specifically to detect the GAL08 16S rRNA gene revealed that absolute and relative abundances of GAL08 peaked at 65°C along three temperature gradients. Analysis of sediment collected over multiple years and locations revealed that the GAL08 group was consistently a dominant clade, comprising up to 29.2% of the microbial community based on relative read abundance and up to 4.7 × 105 16S rRNA gene copy numbers per gram of sediment based on qPCR. Using a medium quality threshold, 25 single amplified genomes (SAGs) representing these bacteria were generated from samples taken at 65 and 77°C, and seven metagenome-assembled genomes (MAGs) were reconstructed from samples collected at 45-77°C. Based on average nucleotide identity (ANI), these SAGs and MAGs represented three separate species, with an estimated average genome size of 3.17 Mb and GC content of 62.8%. Phylogenetic trees constructed from 16S rRNA gene sequences and a set of 56 concatenated phylogenetic marker genes both placed the three GAL08 bacteria as a distinct subgroup of the phylum Acidobacteria, representing a candidate order (Ca. Frugalibacteriales) within the class Blastocatellia. Metabolic reconstructions from genome data predicted a heterotrophic metabolism, with potential capability for aerobic respiration, as well as incomplete denitrification and fermentation. In laboratory cultivation efforts, GAL08 counts based on qPCR declined rapidly under atmospheric levels of oxygen but increased slightly at 1% (v/v) O2, suggesting a microaerophilic lifestyle

Stable Isotope and Metagenomic Profiling of a Methanogenic Naphthalene-Degrading Enrichment Culture

Polycyclic aromatic hydrocarbons (PAH) such as naphthalene are widespread, recalcitrant pollutants in anoxic and methanogenic environments. A mechanism catalyzing PAH activation under methanogenic conditions has yet to be discovered, and the microbial communities coordinating their metabolism are largely unknown. This is primarily due to the difficulty of cultivating PAH degraders, requiring lengthy incubations to yield sufficient biomass for biochemical analysis. Here, we sought to characterize a new methanogenic naphthalene-degrading enrichment culture using DNA-based stable isotope probing (SIP) and metagenomic analyses. 16S rRNA gene sequencing of fractionated DNA pinpointed an unclassified Clostridiaceae species as a putative naphthalene degrader after two months of SIP incubation. This finding was supported by metabolite and metagenomic evidence of genes predicted to encode for enzymes facilitating naphthalene carboxylic acid CoA-thioesterification and degradation of an unknown arylcarboxyl-CoA structure. Our findings also suggest a possible but unknown role for Desulfuromonadales in naphthalene degradation. This is the first reported functional evidence of PAH biodegradation by a methanogenic consortium, and we envision that this approach could be used to assess carbon flow through other slow growing enrichment cultures and environmental samples