In this paper, we present a simple, versatile method that creates patterns for cell migration studies using thermoplastic elastomer (TPE). The TPE material used here can be robustly, but reversibly, bonded to a variety of plastic substrates, allowing patterning of cultured cells in a microenvironment. We first examine the bonding strength of TPE to glass and polystyrene substrates and com-pare it to thermoset silicone-based PDMS under various conditions and demonstrate that the TPE can be strongly and reversibly bonded on commercially available polystyrene culture plates. In cell migration studies, cell patterns are templated around TPE features cored from a thin TPE film. We show that the significance of fibroblast cell growth with fetal bovine serum (FBS)-cell culture media compared to the cells cultured without FBS, analyzed over two days of cell culture. This simple approach allows us to generate cell patterns without harsh manipulations like scratch assays and to avoid damaging th... [more]

Defined by its potential for self-renewal, differentiation and tumorigenicity, cancer stem cells (CSCs) are considered responsible for drug resistance and relapse. To understand the behavior of CSC, the effects of the microenvironment in each tissue are a matter of great concerns for scientists in cancer biology. However, there are many complicated obstacles in the mimicking the microenvironment of CSCs even with current advanced technology. In this context, novel biomaterials have widely been assessed as in vitro platforms for their ability to mimic cancer microenvironment. These efforts should be successful to identify and characterize various CSCs specific in each type of cancer. Therefore, extracellular matrix scaffolds made of biomaterial will modulate the interactions and facilitate the investigation of CSC associated with biological phenomena simplifying the complexity of the microenvironment. In this review, we summarize latest advances in biomaterial scaffolds, which are explo... [more]

In this study, we investigated thermophilic (55 °C) anaerobic digestion (AD) performance and microbial community structure, before and after hydrogen addition, in a novel hybrid gas-stirred tank reactor (GSTR) implemented with a partial immobilization of the microbial community and fed with second cheese whey (SCW). The results showed that H2 addition led to a 25% increase in the methane production rate and to a decrease of 13% in the CH4 concentration as compared with the control. The recovery of methane content (56%) was reached by decreasing the H2 flow rate. The microbial community investigations were performed on effluent (EF) and on interstitial matrix (IM) inside the immobilized area. Before H2 addition, the Anaerobaculaceae (42%) and Lachnospiraceae (27%) families dominated among bacteria in the effluent, and the Thermodesulfobiaceae (32%) and Lachnospiraceae (30%) families dominated in the interstitial matrix. After H2 addition, microbial abundance showed an increase in the ba... [more]

The usage of lignocellulosic biomass in energy production for biofuels and other value-added products can extensively decrease the carbon footprint of current and future energy sectors. However, the infrastructure in the processing of lignocellulosic biomass is not well-established as compared to the fossil fuel industry. One of the bottlenecks is the production of the lignocellulolytic enzymes. These enzymes are produced by different fungal and bacterial species for degradation of the lignocellulosic biomass into its reactive fibers, which can then be converted to biofuel. The selection of an ideal feedstock for the lignocellulolytic enzyme production is one of the most studied aspects of lignocellulolytic enzyme production. Similarly, the fermentation enhancement strategies for different fermentation variables and modes are also the focuses of researchers. The implementation of fermentation enhancement strategies such as optimization of culture parameters (pH, temperature, agitation,... [more]

High removal of heavy metals using plant-based bioflocculant under low concentration is required due to its low cost, abundant source, and nontoxicity for improved wastewater management and utilization in the water industry. This paper presents a treatment of synthetic wastewater using plant-based Tacca leontopetaloides biopolymer flocculant (TBPF) without modification on its structural polymer chains. It produced a high removal of heavy metals (Zn, Pb, Ni, and Cd) at a low concentration of TBPF dosage. In our previous report, TBPF was characterized and successfully reduced the turbidity, total suspended solids, and color for leachate treatment; however, its effectiveness for heavy metal removal has not been reported. The removal of these heavy metals was performed using a standard jar test procedure at different pH values of synthetic wastewater and TBPF dosages. The effects of hydroxide ion, pH, initial TBPF concentration, initial metal ion concentration, and TBPF dosage were examine... [more]

Different antibiotics contained in manure, slurry, wastewater or sewage sludge are spread into the environment. The harmful effects of these antibiotics could be minimized by means of immobilization onto bioadsorbent materials. This work investigates the competitive adsorption/desorption of tetracycline (TC) and sulfadiazine (SDZ) onto pine bark, oak ash and mussel shell. The study was carried out using batch-type experiments in binary systems (with both antibiotics present simultaneously), adding 5 equal concentrations of the antibiotics (between 1 and 50 µmol L−1). The adsorption percentages were higher for TC (close to 100% onto pine bark and oak ash, and between 40 and 85% onto mussel shell) than for SDZ (75−100% onto pine bark, and generally less than 10% on oak ash and mussel shell). Pine bark performed as the best adsorbent since TC adsorption remained close to 100% throughout the entire concentration range tested, while it was between 75 and 100% for SDZ. Desorption was always... [more]

The aim of our study was to obtain rutoside (RUT) inclusion complexes in β-cyclodextrin (β-CD) and in hydroxypropyl-β-cyclodextrin (HP-β-CD), in a 1:1 molar ratio, using the lyophilization method of complexation in solution. The complexes were confirmed and characterized, in comparison with the raw materials and their simple physical mixtures, by SEM, DSC, and FT-IR analyses. The antioxidant activity of the compounds was assessed by using the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and 2’-azino-bis(3-ethylbenzothiazolin-6-sulfonic) acid (ABTS) radicals, determining the radical scavenging activity, and by ferric reducing antioxidant power (FRAP) assay. The results revealed superior antioxidant ability for the inclusion complexes towards rutoside alone. The inclusion complexes were used as active ingredients in formulations of immediate-release tablets. The preformulation studies were performed on the powders for direct compression obtained after mixing the active ingredients with the exci... [more]

In this second part of our systematic review on the research area of 3D cell culture in micro-bioreactors we give a detailed description of the published work with regard to the existing micro-bioreactor types and their applications, and highlight important results gathered with the respective systems. As an interesting detail, we found that micro-bioreactors have already been used in SARS-CoV research prior to the SARS-CoV2 pandemic. As our literature research revealed a variety of 3D cell culture configurations in the examined bioreactor systems, we defined in review part one “complexity levels” by means of the corresponding 3D cell culture techniques applied in the systems. The definition of the complexity is thereby based on the knowledge that the spatial distribution of cell-extracellular matrix interactions and the spatial distribution of homologous and heterologous cell−cell contacts play an important role in modulating cell functions. Because at least one of these parameters ca... [more]

We aimed in the current study, the identification of a marine bacterial amylase produced by Bacillus pacificus, which was associated with Turbinaria ornata. Cultural conditions were optimized for the highest amylase production on Tryptic soy broth media supplemented with starch 1% at initial pH 9, 55 °C for 24 h. The newly purified amylase was characterized for a possible biotechnological application. Data indicated that the obtained amylase with a molecular weight of 40 kD and the N-terminal sequence of the first 30 amino acids of amBp showed a high degree of homology with known alpha amylase, and was stable at 60 °C of pH 11. Among the tested substrate analogs, amBp was almost fully active on Alylose and Alylopectine (97%), but moderately hydrolyzed glycogen < sucrose < maltose < lactose. Therefore, the current amylase mainly generated maltohexaose from starch. Mg2+ and Zn2+ improved amylase activity up to 170%. While ethylenediamine tetraacetic acid (EDTA) similarly induced... [more]

Candida antarctica Lipase B (CALB) has a wide range of applications in many fields. In this study, Pichia pastoris was used to express CALB for fermentation tank culture. Sodium dodecyl sulfate (SDS) was used as a surfactant, and foam separation technology was used to explore the best experimental conditions for the harvest of CALB. The results showed that the optimal technological conditions for the foam separation and recovery of CALB were as follows: liquid volume was 150 mL, separating gas velocity was 600 mL/min, pH value was 7, and surfactant SDS concentration was 0.5 mg/mL. Under these conditions, the enrichment ratio of CALB was 0.95, and recovery rate R was 80.32%, respectively, indicating that the foam separation technology is feasible to extract lipase B.

Regenerative medicine is a novel scientific field that employs the use of stem cells as cell-based therapy for the regeneration and functional restoration of damaged tissues and organs. Stem cells bear characteristics such as the capacity for self-renewal and differentiation towards specific lineages and, therefore, serve as a backup reservoir in case of tissue injuries. Therapeutically, they can be autologously or allogeneically transplanted for tissue regeneration; however, allogeneic stem cell transplantation can provoke host immune responses leading to a host-versus-transplant reaction. A probable solution to this problem is stem cell encapsulation, a technique that utilizes various biomaterials for the creation of a semi-permeable membrane that encases the stem cells. Stem cell encapsulation can be accomplished by employing a great variety of natural and/or synthetic hydrogels and offers many benefits in regenerative medicine, including protection from the host’s immune system and... [more]

Several microalgae species have been exploited due to their great biotechnological potential for the production of a range of biomolecules that can be applied in a large variety of industrial sectors. However, the major challenge of biotechnological processes is to make them economically viable, through the production of commercially valuable compounds. Most of these compounds are accumulated inside the cells, requiring efficient technologies for their extraction, recovery and purification. Recent improvements approaching physicochemical treatments (e.g., supercritical fluid extraction, ultrasound-assisted extraction, pulsed electric fields, among others) and processes without solvents are seeking to establish sustainable and scalable technologies to obtain target products from microalgae with high efficiency and purity. This article reviews the currently available approaches reported in literature, highlighting some examples covering recent granted patents for the microalgae’s compone... [more]

Effective mosquito monitoring relies on the accurate identification and characterization of the target population. Since this process requires specialist knowledge and equipment that is not widely available, automated field-deployable systems are highly desirable. We present a centrifugal microfluidic cartridge, the VectorDisk, which integrates TaqMan PCR assays in two feasibility studies, aiming to assess multiplexing capability, specificity, and reproducibility in detecting disk-integrated vector-related assays. In the first study, pools of 10 mosquitoes were used as samples. We tested 18 disks with 27 DNA and RNA assays each, using a combination of multiple microfluidic chambers and detection wavelengths (geometric and color multiplexing) to identify mosquito and malaria parasite species as well as insecticide resistance mechanisms. In the second study, purified nucleic acids served as samples to test arboviral and malaria infective mosquito assays. Nine disks were tested with 14 as... [more]

Hemophilia, a congenital X linked disease, has the serious burden of bleeding, requiring life-long replacement with coagulation factors (CF). In the present day, there is a continuously improving treatment for this condition. Objective: Our observational, cross-sectional study aims at finding out whether a prophylactic replacement with CF is affordable from the point of view of its cost-effectiveness in our country. Material and methods: A cohort of 122 persons with hemophilia were included in this patient-reported outcome survey, and they answered a questionnaire consisting of 56 items, focused on 4 domains—socio-demographic, medical, quality of health/life and cost/cost-effectiveness. Results and discussion: The markers for quality of health/life were correlated with the direct and indirect costs of care, comparing subgroup 1 of patients with prophylactic vs. subgroup 2 with on-demand replacement. Based on the incremental quality adjusted life years and the incremental costs, we calc... [more]

The present study focuses on the inhibitory effect of volatile metabolites released by Bacillus velezensis CT32 on Verticillium dahliae and Fusarium oxysporum, the causal agents of strawberry vascular wilt. The CT32 strain was isolated from maize straw compost tea and identified as B. velezensis based on 16S rRNA gene sequence analysis. Bioassays conducted in sealed plates revealed that the volatile organic compounds (VOCs) produced by the strain CT32 possessed broad-spectrum antifungal activity against eight phytopathogenic fungi. The volatile profile of strain CT32 was obtained by headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS). A total of 30 volatile compounds were identified, six of which have not previously been detected in bacteria or fungi: (Z)-5-undecene, decyl formate, 2,4-dimethyl-6-tert-butylphenol, dodecanenitrile, 2-methylpentadecane and 2,2’,5,5’-tetramethyl-1,1’-biphenyl. Pure compounds were tested in vitro for th... [more]

Radiofrequency (RF)-based monopolar (MM) and bipolar mode (BM) applicators are used to thermally create coagulation zones (CZs) in biological tissues with the aim of destroying surface tumors and minimizing blood losses in surgical resection. Both modes have disadvantages as regards safely and in obtaining a sufficiently deep coagulation zone (CZ). In this study, we compared both modes versus a switching monopolar mode (SMM) in which the role of the active electrode changes intermittently between the two electrodes of the applicator. In terms of clinical impact, the three modes can easily be selected by the surgeon according to the surgical maneuver. We used computational and experimental models to study the feasibility of working in MM, BM, and SMM and to compare their CZ characteristics. We focused exclusively on BM and SMM, since MM only creates small coagulation zones in the area between the electrodes. The results showed that SMM produces the deepest CZ between both electrodes (33... [more]

Bioreactors have proven useful for a vast amount of applications. Besides classical large-scale bioreactors and fermenters for prokaryotic and eukaryotic organisms, micro-bioreactors, as specialized bioreactor systems, have become an invaluable tool for mammalian 3D cell cultures. In this systematic review we analyze the literature in the field of eukaryotic 3D cell culture in micro-bioreactors within the last 20 years. For this, we define complexity levels with regard to the cellular 3D microenvironment concerning cell−matrix-contact, cell−cell-contact and the number of different cell types present at the same time. Moreover, we examine the data with regard to the micro-bioreactor design including mode of cell stimulation/nutrient supply and materials used for the micro-bioreactors, the corresponding 3D cell culture techniques and the related cellular microenvironment, the cell types and in vitro models used. As a data source we used the National Library of Medicine and analyzed the s... [more]

Elementary Flux Modes (EFMs) provide a rigorous basis to systematically characterize the steady state, cellular phenotypes, as well as metabolic network robustness and fragility. However, the number of EFMs typically grows exponentially with the size of the metabolic network, leading to excessive computational demands, and unfortunately, a large fraction of these EFMs are not biologically feasible due to system constraints. This combinatorial explosion often prevents the complete analysis of genome-scale metabolic models. Traditionally, EFMs are computed by the double description method, an efficient algorithm based on matrix calculation; however, only a few constraints can be integrated into this computation. They must be monotonic with regard to the set inclusion of the supports; otherwise, they must be treated in post-processing and thus do not save computational time. We present aspefm, a hybrid computational tool based on Answer Set Programming (ASP) and Linear Programming (LP) th... [more]

Continuous manufacturing of biologics (biopharmaceuticals) has been an area of active research and development for many reasons, ranging from the demand for operational streamlining to the requirement of achieving obvious economic benefits. At the same time, biopharma strives to develop systems and concepts that can operate at similar scales for clinical and commercial production—using flexible infrastructures, such as single-use flow paths and small surge vessels. These developments should simplify technology transfer, reduce footprint and capital investment, and will allow to react readily to changing market pressures while maintaining quality attributes. Despite a number of clearly identified benefits compared to traditional batch processes, continuous bioprocessing is still not widely adopted for commercial manufacturing. This paper details how industry-specific technological, organizational, economic, and regulatory barriers that exist in biopharmaceutical manufacturing are hinder... [more]

Microalgae cultivation in liquid foams is a promising concept which requires the use of a surfactant as a foam stabilizing agent. The biodegradable character of a surfactant is a key aspect regarding its applicability in a liquid foam-bed photobioreactor (LF-PBR), since it might influence microalgal growth and the stability of the foam-based cultivation. In this work, the effects of the biodegradable surfactants bovine serum albumin (BSA), Saponin and Tween 20 on the whole microbial community of microalgal cultures (i.e., microalgal and bacterial populations) were studied. The three surfactants enhanced bacterial and microalgal growth in non-axenic microalgal cultures, but they differed in their efficiency to sustain bacterial growth. In this sense, Saponin was proven to enhance the growth of S. obliquus-associated bacteria in microalgae-free cultures, and to sustain it even when other nutrients were lacking, suggesting that Saponin can be used as an energy and nutrients source by thes... [more]

Blood is not always available in forensic autopsies, therefore, the search for alternative sampling materials is needed. This study aimed at examining if ethanol can be detected in costal cartilage and to investigate if different forms of costal cartilage can give accurate information about ethanol concentration in the blood or urine of human cadavers (n = 50). Ethanol concentration in samples of unground costal cartilage (UCC), ground costal cartilage (GCC), femoral venous blood, and urine was analyzed using a gas chromatography-flame ionization detector (GC-FID). Due to Polish law, we used two different cut-off points: the blood alcohol concentration >0.2 mg/mL defined as the ‘after use’ condition, and the blood alcohol concentration >0.5 mg/mL defined as the ‘state of insobriety’. Based on the constructed receiver operating characteristics (ROC) curves, the optimal cut-off point for ethanol content as the ‘after use’ condition was 0.273 mg/g for the UCC method and 0.069 mg/g for the... [more]

The development of an advanced and efficient drug delivery system with significant improvement in its efficacy and enhanced therapeutic value is one of the critical challenges in modern medicinal biology. The integration of nanomaterial science with molecular and cellular biology has helped in the advancement and development of novel drug delivery nanocarrier systems with precision and decreased side effects. The design and synthesis of nanocarriers using graphene oxide (GO) have been rapidly growing over the past few years. Due to its remarkable physicochemical properties, GO has been extensively used in efforts to construct nanocarriers with high specificity, selectivity, and biocompatibility, and low cytotoxicity. The focus of this review is to summarize and address recent uses of GO-based nanocarriers and the improvements as efficient drug delivery systems. We briefly describe the concepts and challenges associated with nanocarrier systems followed by providing critical examples of... [more]

Liposomes are bilayer membrane vesicles that can serve as vehicles for drug delivery. They are a good alternative to free drug administration that provides cell-targeted delivery into tumors, limiting the systemic toxicity of chemotherapeutic agents. Previous results from our group showed that an astrocytoma cell line exhibits selective uptake of sulfatide-rich (SCB) liposomes, mediated by the low-density lipoprotein receptor (LDL-R). The goal of this study was to assess the uptake of liposomes in a neuroblastoma cell line. For this purpose, we used two types of liposomes, one representing a regular cell membrane (DOPC) and another rich in myelin components (SCB). An astrocytoma cell line was used as a control. Characterization of liposome uptake and distribution was conducted by flow cytometry and fluorescence microscopy. Similar levels of LDL-R expression were found in both cell lines. The uptake of SCB liposomes was higher than that of DOPC liposomes. No alterations in cell viabilit... [more]

Protein phosphorylation is the most widespread type of post-translational modification and is properly controlled by protein kinases and phosphatases. Regarding the phosphorylation of serine (Ser) and threonine (Thr) residues, relatively few protein Ser/Thr phosphatases control the specific dephosphorylation of numerous substrates, in contrast with Ser/Thr kinases. Recently, protein Ser/Thr phosphatases were reported to have rigid substrate recognition and exert various biological functions. Therefore, identification of targeted proteins by individual protein Ser/Thr phosphatases is crucial to clarify their own biological functions. However, to date, information on the development of methods for identification of the substrates of protein Ser/Thr phosphatases remains scarce. In turn, substrate-trapping mutants are powerful tools to search the individual substrates of protein tyrosine (Tyr) phosphatases. This review focuses on the development of novel methods for the identification of S... [more]

A large portion of annual deaths worldwide are due to infections caused by disease-causing pathogens. These pathogens contain virulence genes, which encode mechanisms that facilitate infection and microbial survival in hosts. More recently, antimicrobial resistance (AMR) genes, also found in these pathogens, have become an increasingly large issue. While the National Center for Biotechnology Information (NCBI) Pathogen Detection Isolates Browser (NPDIB) database has been compiling genes involved in microbial virulence and antimicrobial resistance through isolate samples, few studies have identified the genes primarily responsible for virulence and compared them to those responsible for AMR. This study performed the first multivariate statistical analysis of the multidimensional NPDIB data to identify the major virulence genes from historical pathogen isolates for Australia, China, South Africa, UK, and US—the largely populated countries from five of the six major continents. The import... [more]