Control Cultures Research Articles

Control of Tissue Strain Is Essential for Enhanced Dermal Innervation in the Three-Dimensional Skin Engineering.

Engineered skin models with sensory innervation are a growing and challenging field of research aimed at applications in regenerative medicine, biosensing, and drug screening. Researchers are attempting to fabricate innervated skin tissues using collagen sponges, cell culture inserts, and microfluidic devices to partially mimic the layered structure of the skin. However, innervation of the full-thickness skin model has not yet been achieved. Here, using the anchoring culture device we previously reported, which is a powerful tool to construct a full-thickness three-dimensional (3D) skin model with balanced tissue contraction forces, we drastically improved dermal layer innervation using a composite hydrogel of collagen and Matrigel (Coll:MG). To determine the preferable hydrogel matrix for neurite extension in the 3D skin construct, DRG neural spheroids were placed at the bottom of the dermal layer composed of various hydrogel scaffold, including type I collagen from different origins (dermis or tendon) and Coll:MG composite hydrogel with different compositions. We showed that the Coll:MG (2:1) composite hydrogel significantly increased vertical neurite extension in the dermal layer, concomitant with the reduced tissue shrinkage during the culture. In contrast, in the collagen-only hydrogel, neurite extension occurred mostly in the horizontal direction, and tissues sometimes detached from the anchors due to significant shrinkage, indicating that tissue shrinkage may affect the direction of neurite extension. To exemplify this idea, 3D skin constructed in the device was partially detached from the anchors to comply with the cell-induced tissue shrinkage and reduce the strain on the tissue. The data showed that the partial allowance of in-plane tissue strain remarkably increased vertical neurite extension compared to the control cultures. Collectively, our results strongly suggest that neurite extension angles can be modulated by adjusting the tissue strain during the culture. Our findings highlight the importance of controlling tissue strain for the advancement of an innervated skin model.

Controlled Dynamic Microfluidic Culture of Murine, Bovine, and Human Embryos Improves Development: Proof-of-Concept Studies

Classical preimplantation embryo culture is performed in static fluid environments. Whether a dynamic fluid environment, like the fallopian tube, is beneficial for embryo development remains to be determined across mammalian species. Objectives of these proof-of-concept studies were to determine if controllable dynamic microfluidic culture would enhance preimplantation murine, bovine, and human embryo development compared to static culture. This prospective randomized controlled trial tested static versus controlled dynamic culture of preimplantation mouse (n = 397), bovine (n = 242), and human (n = 512) zygotes to blastocyst stages with outcome measures of embryo cleavage, cellular fragmentation, apoptosis, and blastocyst conversion rates. Dynamic culture of mouse and bovine zygotes with microfluidics significantly improved embryo development. Mouse placental imprinted gene expression was significantly different between embryos derived in vivo, by static culture, and by dynamic culture. Using human sibling zygotes, this dynamic microfluidic culture system increased the number of blastomeres per cleavage-stage embryo, reduced cellular fragmentation or apoptosis, improved blastocyst conversion rates, and enhanced blastocyst developmental stages. In conclusion, species-specific longitudinal studies demonstrated that dynamic microfluidic culture significantly improved embryo development, independent of culture media composition, temperature, and gaseous environment. These cellular indicators represent improved embryo development that can translate into higher pregnancy rates in transgenics, domestic livestock and endangered species and treating human infertility.

Navigating readiness for change: exploring the influence of authentic leadership, culture, learning and internal locus of control

Navigating readiness for change: exploring the influence of authentic leadership, culture, learning and internal locus of control

The mediating effect of patient safety culture on the relationship between professionalism, self-leadership, and compliance with infection prevention activity against emerging respiratory infectious disease among nurses working in geriatric long-term care hospitals: A cross-sectional study

Purpose: This study investigates the mediating effect of patient safety culture on the relationship between professionalism, self-leadership, and compliance with infection prevention activity against emerging respiratory infectious diseases among nurses working in geriatric long-term care hospitals. Methods: In total, 136 nurses from eight geriatric long-term care hospitals were invited to participate. For the statistical analyses, descriptive statistics, t-test, ANOVA, and Pearson’s correlation coefficient analyses were employed. The mediating effect was analyzed using the three-step mediation procedure of Baron and Kenny in hierarchical regression, and the significance of the mediating effect was tested using the Sobel test. Results: Patient safety culture had a full mediating effect on the relationship between professionalism and compliance with infection prevention activity (Z=3.14, p=.001) and a full mediating effect on the relationship between self-leadership and compliance with infection prevention activity (Z=3.37, p<.001). Conclusion: The study findings indicate that a higher patient safety culture within the organization further enhances compliance with infection prevention activity. Therefore, it is crucial to develop strategies to enhance nurses’ professionalism and self-leadership, as well as to improve organizational patient safety culture for the infection control in geriatric long-term care hospitals.

Pericytes require physiological oxygen tension to maintain phenotypic fidelity

Pericytes function to maintain tissue homeostasis by regulating capillary blood flow and maintaining endothelial barrier function. Pericyte dysfunction is associated with various pathologies and has recently been found to aid cancer progression. Despite having critical functions in health and disease, pericytes remain an understudied population due to a lack of model systems which accurately reflect in vivo biology. In this study we developed a protocol to isolate and culture murine lung, brain, bone, and liver pericytes, that maintains their known phenotypes and functions. We demonstrate that pericytes, being inherently plastic, benefit from controlled oxygen tension culture conditions, aiding their expansion ex vivo. Primary pericytes grown in physiologically relevant oxygen tensions (10% O2 for lung; 5% O2 for brain, bone, and liver) also better retain pericyte phenotypes indicated by stable expression of characteristic transcriptional and protein markers. In functional tube formation assays, pericytes were observed to significantly associate with endothelial junctions. Importantly, we identified growth conditions that limit expression of the plasticity factor Klf4 to prevent spontaneous phenotypic switching in vitro. Additionally, we were able to induce pathological pericyte phenotypic switching in response to metastatic stimuli to accurately recapitulate in vivo biology. Here, we present a robust method for studying pericyte biology in both physiology and disease.

Influence of neodymium-doped yttrium aluminum garnet laser exposure time on cytokine secretion in lipopolysaccharide-challenged rat peripheral blood mononuclear cells

Multiple investigators have suggested that infrared laser energy facilitates hard and soft tissue wound healing through various mechanisms, including the suppression of inflammation. This study investigated the influence of neodymium-doped yttrium aluminum garnet (Nd:YAG) laser exposure time on pro-inflammatory cytokine/chemokine concentrations in lipopolysaccharide (LPS)-stimulated rat peripheral blood mononuclear cells (PBMCs). Cultured rat PBMCs were stimulated with various LPS concentrations (0, 10, 100, or 1000 ng/mL) and treated with Nd:YAG laser irradiation for 0 (control), 30, 45, or 60 s. For these experiments, the average power, pulse duration, and repetition rate remained constant at 5 W, 100 μs, and 20 Hz, respectively. Luminex magnetic microsphere immunoassays were used to compare the secretion of 27 inflammatory mediators from LPS-stimulated PBMCs in laser-irradiated versus control groups. Two-way analysis of variance was used to compare the main effects of laser exposure time and LPS concentration on cytokine/chemokine concentrations and evaluate the potential interaction between these factors. Four pro-inflammatory cytokines – tumor necrosis factor-α, macrophage inflammatory protein (MIP)-1α, MIP-2, and interferon gamma-induced protein-10 – exhibited a trend of reduced secretion in laser-irradiated cultures. The effect appeared more pronounced at longer exposure times (45 and 60 s). However, none of the 27 inflammatory mediators exhibited statistically significant reductions in concentrations in laser-irradiated cultures versus control cultures. These observations do not support a robust anti-inflammatory effect of Nd:YAG laser irradiation. Further studies should explore the potential impact of Nd:YAG laser irradiation on cytotoxicity and cellular growth kinetics and extend across a range of irradiation parameters and cell types.

Weaving potentialities and AI:

Ngaa tuku o Maahina is a roopuu [group] whose kaupapa [purpose] relates to the multiple interconnected strands of knowledge of ngaa maramataka [site specific lunar calendars]. We are Hagen Tautari, Horomona Horo, Te Taima Barrett, Hollie Tawhiao, Toni Herangi, Ra Keelan and Joe Citizen. At our inaugural hui [gathering], Toni brought along a taonga [prized treasure] which had been in her family for generations – dated 1898, this photolithographed collection in te reo Māori [the Māori language] and English, has copies of He Whakaputanga (Declaration of independence), Te Tiriti o Waitangi (Treaty of Waitangi) and other associated papers. To some, this is might appear to simply be copies of older original documents. In te ao Māori [the Māori world], however, this collection has its own mauri [life-essence], mana [spiritual authority and prestige] and haa [breath] and retains links to the tuupuna [ancestors] through their tohu [signs]. Our koorero [discussion] became centred on its preservation – simply photographing it and uploading copies of it online would make it vulnerable to AI that employ web-scraping and web-crawling practices, to thieve knowledge and assimilate it into a supposedly universalist framework. This koorero brought to our attention the wider take [subject] of how we should navigate the colonial tendencies of AI – the structural violence that arises from those culturally specific practices that assume a priori truths as universally foundational. By appraising the Eurocentric and human-centric assumptions behind data recognition through the automation of iterative logic loops, we acknowledge the entanglements of historically systemic problems: how cognitivism and positivism underpin supremacist claims of liberatory technological progress; how material realism and representational indexicality have traditionally dismissed and abused indigenous knowledge frameworks; how military-industrial control culture has normalised surveillance and transnational capitalism; and how reductionist methodologies reify libertarian claims of individual freedoms as commodified transactions between atomistic entities.

Research note: Increased lipid accumulation within broiler preadipocytes during differentiation in vitro at atmospheric oxygen tension

In the broiler industry, intensive genetic selection has been placed on muscle growth which has undesirably led to increased fat accretion. Models of chicken preadipocyte differentiation in vitro have conventionally used incubators without the ability to control oxygen (O2) tension; thus, the cells are exposed to atmospheric (∼20-21%) O2, which is supraphysiological compared to the O2 tension within adipose tissue. The objective of this study was to investigate embryonic broiler preadipocyte differentiation at different O2 tensions, including atmospheric (20%), physiological (5%), and hypoxic (1%). Culture at 1% O2 resulted in increased abundance of HIF1α, a canonical protein stabilized during hypoxia, thus confirming effectiveness of the treatment. Increased accumulation of lipid was observed in preadipocytes cultured in adipogenic differentiation medium compared to the control medium. When considering oxygen tension, lipid accumulation was increased in preadipocytes that were cultured in differentiation medium at 20% O2 compared to 5% or 1% O2. Furthermore, abundance of transcripts related to fatty acid transport and adipogenesis, fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor gamma (PPARγ), were increased in differentiated preadipocytes cultured at 20% O2 compared to 5% or 1% O2. Abundance of transcripts related to lipid synthesis and oxidation, acyl-CoA synthetase long chain family member 1 (ACSL1) and carnitine palmitoyltransferase 1A (CPT1A), were increased in the differentiation cultures compared to the control cultures. Abundance of glutathione peroxidase 4 (GPX4) was increased in all the differentiation cultures compared to the controls, regardless of oxygen tension; however, differences in the abundance of other antioxidant enzymes were not observed. Overall, exposure to atmospheric oxygen tension promotes lipid accumulation within chicken preadipocytes, which may need to be considered when developing in vitro models of this process.

Improving the biodiesel production in the marine diatom Thalassiosira pseudonana cultivated in nutrient deficiency and sewage water.

The use of microalgae as a feedstock in biofuel production is highly encouraging. The marine diatom in this study, Thalassiosira pseudonana, was used as a test organism to evaluate the impact of nitrogen or phosphorus limitation and sewage water on improving biodiesel production. The growth rate is more affected in cultures without phosphorus by 41.8% lower than in control and the highest dry weight estimated in control. The cellular dry weight significantly increased in cultures treated with mix1 and mix2 wastewater compared to the control cultures. Chlorophyll a content decreased in the absence of nitrogen and phosphorous and in sewage water cultures. Lipid content in algal cultures without nitrogen or phosphorus and in sewage water accumulated nearly twice as much lipid content in synthetic medium. T. pseudonana showed high FAME contents; the two most abundant fatty acids, stearic acid (C18:0) and palmitoleic acid (C16:1), in the algal extracts revealed that T. pseudonana was predominantly composed of these fatty acids. T. pseudonana grown in nitrogen or phosphorus-deficient conditions exhibited an extreme percentage of saturated fatty acids (SFAs) by 87.38% and 85.47%, respectively, of the total fatty acids (TFAs). More importantly, the low polyunsaturated fatty acid content in oils indicates a high cetane number, low iodine value, and low corrosion for biodiesel quality indicators. Producing biodiesel that closely meets worldwide biodiesel requirements (ASTM D6751 and EN 14214) is the goal of this work, which shows that growing T. pseudonana under nutrient limitations leads to algal metabolism in that direction.

Thermoresponsive Brush Coatings for Cell Sheet Engineering with Low Protein Adsorption above the Polymers' Phase Transition Temperature.

Thermoresponsive polymer coatings on cell culture substrates enable noninvasive cell detachment and cell sheet fabrication for biomedical applications. Optimized coatings should support controlled culture and detachment of various cell types and allow chemical modifications, e.g., to introduce specific growth factors for enhanced gene expression. Furthermore, the sterilization and storage stability of the coatings must be assessed for translational attempts. Poly(glycidyl ether) (PGE) brush coatings with short alkoxy side chains provide a versatile platform for cell culture and detachment, but their polyether backbones are susceptible to oxidation and degradation. Thus, we rationally designed potential alternatives with thermoresponsive glycerol-based block copolymers comprising a stable polyacrylate or polymethacrylate backbone and an oligomeric benzophenone (BP)-based anchor. The resulting poly(ethoxy hydroxypropyl acrylate-b-benzophenone acrylate) (pEHPA-b-BP) and poly(ethoxy hydroxypropyl methacrylate-b-benzophenone methacrylate) (pEHPMA-b-BP) block copolymers preserve the short alkoxy-terminated side chains of the PGE derived structure on a stable, but hydrophobic, aliphatic backbone. The amphiphilicity balance is maintained through incorporated hydroxyl groups, which simultaneously can be used for chemical modification. The polymers were tailored into brush coatings on polystyrene surfaces via directed adsorption using the BP oligomer anchor. The resulting coatings with thickness values up to ∼3 nm supported efficient adhesion and proliferation of human fibroblasts despite minimal protein adsorption. The conditions for cell sheet fabrication on pEHPA-b-BP were gentler and more reliable than on pEHPMA-b-BP, which required additional cooling. Hence, the stability of pEHPA-b-BP and PGE coatings was evaluated post gamma and formaldehyde (FO) gas sterilization. Gamma sterilization partially degraded PGE coatings and hindered cell detachment on pEHPA-b-BP. In contrast, FO sterilization only slowed detachment on PGE coatings and had no adverse effects on pEHPA-b-BP, maintaining their efficient performance in cell sheet fabrication.

Alterations in nitrogen metabolism caused by heavy metals in the acid-tolerant microalga Coccomyxa onubensis

The microalga Coccomyxa onubensis is an extremophile microorganism with a unique ecosystem (Río Tinto, Huelva, Spain) that contains high amounts of contaminants, including heavy metals, sulphates, and nitrates, in acidic environments (pH 2.5). The current work presents an evaluation of the capacity of Coccomyxa onubensis to assimilate different nitrogen sources under Cu2+, Cd2+, AsO33−, AsO43− and Hg2+ stress, and the metabolic implications of these stressors. The results showed that ammonium consumption was less affected than nitrate consumption when microalgae were cultivated with heavy metals (except cadmium). The activities of enzymes involved in nitrogen metabolism, such as nitrite reductase (NiR; EC:1.7.7.1), glutamine synthetase (GS; EC:6.3.2.1) and glutamate dehydrogenase (GDH; EC:1.4.1.2) were characterised to determine the Michaelis-Menten constant (Km) and optimal temperature and pH values, being 45, 40 and 60 °C and pH values of 7.5, 6.0 and 9.0 for NiR, GS, and GDH, respectively. The effects of different heavy metals on these enzymes were assessed at multiple levels, and the results showed that the enzymatic activity of NiR was downregulated, specially under copper stress, maintaining 23 % of control NiR activity at 2 mM Cu2+. The enzymatic activity of GS was upregulated at low concentrations under cadmium and mercury stress (115–120 % of control cultures GS activity at 25 μM Cd2+ and 50 nM Hg2+, respectively) and downregulated at high concentrations of these elements. GDH activity was upregulated in the presence of Cu2+, Cd2+, and Hg2+, with increases up to 192, 155 and 154 % at 1 mM Cu2+, 300 μM Cd2+, and 250 nM Hg2+, respectively. These results provide a better explanation of the effects of heavy metal stress on N metabolism in Coccomyxa onubensis, which may be used as a model eukaryotic organism of the Tinto River acidophilic ecosystem.

Growth of Picochlorum celeri in produced water from the Permian Basin (US)

Produced water generated from hydraulic fracking is a waste stream that, on one hand, demands remediation, and, on the other hand, can be used as a water source to support a circular economy. In this study, we aimed to assess the ability of Picochlorum celeri to grow in two different sources of raw, untreated produced water collected in New Mexico, USA. The produced water had initial salinities of 110 and 140 PPT. Picochlorum was screened, with additional algae, on two different sources of produced water supplemented with nitrogen and phosphorus. The strain was able to grow on 25 % and 50 % produced water and achieve biomass densities between ~40 and 100 % of those demonstrated by control cultures. To separate the effects of high salinity stress from toxicity of the produced water samples, we conducted a bioassay with produced water under isotonic conditions of 60 PPT. When controlling for salinity, the contaminants associated with the two sources of produced water did not affect growth of the algae. This study provides justification to consider P. celeri as a candidate for remediation of and biomass production on produced water. Foundational studies are necessary to assess remediation potential and characterize the growth, biomass accumulation, and biomass composition of P. celeri grown on a variety of produced water streams.

Dual drug delivery systems based on natural and synthetic polymer particles for isoniazid and rifampicin vehiculization

AbstractThe purpose of this work is the design and characterization of novel systems based on micro and nanoparticles prepared from poly(lactic‐co‐glycolic) acid (PLGA), chitosan (CHT) and gelatin (GEL) for compartmentalization and dual release of rifampicin and isoniazid. The emulsion/evaporation method is employed for the preparation of PLGA based microparticles and nanoparticles. CHT and GEL are used for the preparation of microparticles by spray‐drying. Entrapment efficiencies are in the range of 35%–73% and 5%–29% for rifampicin and isoniazid in PLGA microparticles, respectively. For isoniazid GEL:CHT particles the entrapment efficiencies are within 82%–100%. Isoniazid shows a complete release at acid condition from GEL:CHT particles, while rifampicin release is maintained from GEL:CHT particles or displays an slightly increase from PLGA particles during buffer conditions. Antimycobacterial activity experiments show that the formulations present the ability to inhibit the bacterial growth in comparison with a control culture without treatment. Rifampicin PLGA and isoniazid GEL:CHT microparticles could be used for the preparation of a dual drug delivery system with each antibiotic in a single particulate compartment. GEL:CHT microparticles containing free isoniazid and rifampicin PLGA nanoparticles are a novel two compartment delivery system.

Bacterial endophyte Pseudomonas mosselii PR5 improves growth, nutrient accumulation, and yield of rice (Oryza sativa L.) through various application methods

BackgroundPseudomonas spp. have drawn considerable attention due to their rhizospheric abundance and exceptional plant growth-promoting attributes. However, more research is needed on the optimal application methods of Pseudomonas mosselii for rice growth, nutrient accumulation, and yield improvement. This research explored the application of the endophytic bacterium P. mosselii PR5 on rice cultivar BRRI dhan29 with four treatments: control, seedling priming, root drenching, and bacterial cell-free culture (CFC) foliar application.ResultsPR5 led to better rice growth, improved nutrient acquisition, and higher yields compared to the control, regardless of the application method used. The highest results in fresh weight of root (146.93 g/pot), shoot (758.98 g/pot), and flag leaf (7.88 g/pot), dry weight of root (42.16 g/pot), shoot (97.32 g/pot), and flag leaf (2.69 g/pot), and grains/panicle (224.67), were obtained from seedling priming treatment, whereas root drenching resulted in maximum plant height (105.67 cm), root length (49.0 cm), tillers/pot (23.7), and panicles/pot (17.67). In all three application methods, rice grain yield per pot was higher in PR5 inoculated treatments, compared to the control. The amount of P, Mg and Zn in the shoot and N, P, Ca, Mg and Si content in the flag leaf was significantly increased along with effective suppression of naturally occurring blast disease in bacterial CFC foliar application, validated by multivariate analysis.ConclusionOur results indicated that rice seedlings priming with PR5 improved rice growth, yield and nutrient uptake, whereas CFC foliar application significantly increased the concentration of most nutrients in the rice plant and suppressed the naturally occurring rice blast disease. This research highlights the significant potential of P. mosselii PR5 in enhancing rice growth, yield, and nutrient uptake, particularly through seedling priming and CFC foliar application methods.

Serum-free Quality and Quantity Control Culture Improves the Angiogenic Potential of Peripheral Blood Mononuclear Cells Harvested from Patients with Connective Tissue Diseases

Serum-free Quality and Quantity Control Culture Improves the Angiogenic Potential of Peripheral Blood Mononuclear Cells Harvested from Patients with Connective Tissue Diseases

A Dot-Blot Screening for Identifying the Temozolomide-Regulated Proteins as Potential Targets for Glioma Multi-OMICS Studies.

Malignant gliomas represent a heterogenous group of brain cancers that are characterized by infiltrative growth that lacks a clearly identifiable tumor border. The lack of the possibility of radical surgical resection and targeted therapy results in a poor prognosis. Although Temozolomide (TMZ) is still the leading chemotherapeutic agent in glioma treatment, its efficacy is limited due to the development of tumor resistance. Therefore, there an urgent need to improve the diagnosis and treatment of these tumors. Finding and developing biomarkers that are specific to glioma could be useful for both identifying therapy targets and monitoring treatment as well as for constructing a personalized therapy. However, there are still no reliable markers that would change the quality of glioma treatment. In this study, differences in the expression of 84 cancer-related proteins in three glioma cell lines were analyzed using the dot-blot method: commercial T98G cells and two patient-derived cell lines. The influence of TMZ on changes in protein expression, cell morphology, and migration was also investigated (Proteome Profiler Human XL Oncology Array, LeviCell System, Microscopic imaging). The lines that were analyzed were characterized by a remarkably different plasticity of protein expression and the proteomic alterations that were induced by TMZ. A dot-blot analysis revealed ten proteins that were common to all of the lines and five (Cathepsin b, FGF, Survivin, AXL, Osteopontin) that were modulated by the TMZ. As a result of the exposure of TMZ, the proteins that are involved in chemoresistance and invasion (TIE-2, Thrombospondin) were detected in both the HROG02 and T98G cell lines. In the control culture (not exposed to TMZ) of HROG17 cells, the proteins that are involved in metabolism were strongly suppressed. The presented data sheds new light on the modulatory effect of Temozolomide on the expression of a protein panel: Cathepsin b, fibroblast growth factor (FGF), Survivin, AXL, and Osteopontin that may suggest their potential as therapeutic targets or biomarkers to monitoring therapy effects. However, further high-throughput analysis and detection of the proteins in the body fluids are necessary.

Studying the Antioxidant and Antimicrobial Activity of Lipsticks Made with Phycoerythrin Under the Influence of Different Magnetic Fields

Background: Several studies have shown the benefits of magnetic treatment on the productivity of secondary metabolites, growth, and the state of microalgae cultures Objective: This study examined the antioxidant and antimicrobial activity of lipsticks made with phycoerythrin (PE) extracted from the cyanobacterium Nostoc sp. under the influence of different magnetic fields (30 mT and 60 mT). Material and Methods: After cyanobacterial culture under magnetic fields of 30 mT and 60 mT, the PE was extracted and lipsticks were formulated. The primary evaluation methods used in this study are melting point, breaking point, linoleic acid peroxidation assay, force of application, stability, surface abnormalities, skin irritation, thixotropy character, dispersibility test, perfume stability, colorimetric assay, antioxidant, and microbial analysis Results: The 30 mT treatment showed the highest concentration, purity, dry weight, antioxidant activity, and percentage of PE extraction compared to control cultures. No significant differences were found in the melting point, stability, thixotropy character, dispersibility, or perfume stability tests. The breaking point and force of application decreased significantly during 30 days. Peroxidation assay tests revealed lipstick increased oxidation and antioxidant activity after 30 days of 30 and 60 mT treatments compared to non-PE cultures. The study found that the amount of ΔE increased significantly in cultures without PE over time, while this increase was lower in magnetic field-treated cultures. However, no signs of crystal formation, surface wrinkles, liquid secretion, itching, or skin irritation were observed in 30 days of 30 and 60 mT magnetic treatments compared to control cultures with PE. Microbial analyzes over 30 days showed a significantly lower number of bacteria under magnetic fields than control cultures. In addition, the results of counting Escherichia coli and coliform bacteria were negative for thirty days. The antioxidant activity of PE was significantly higher in magnetic field-treated cultures. The number of Staphylococcus aureus decreased significantly in all cultures under magnetic field influence. Conclusions: The overall results of this study showed that magnetic fields had a significant effect in many evaluation tests on the culture of cyanobacteria Nostoc sp. As a result, lipsticks made with extracted PE have more antioxidant and antimicrobial activity

Omics exploration of Tetraselmis chuii adaptations to diverse light regimes.

Microalgae are significantly influenced by light quality and quantity, whether in their natural habitats or under laboratory and industrial culture conditions. The present study examines the adaptive responses of the marine microalga Tetraselmis chuii to different light regimes, using a cost-effective filtering method and a multi-omics approach. Microalgal growth rates were negatively affected by all filtered light regimes. After six days of cultivation, growth rate for cultures exposed to blue and green filtered light was 67%, while for red filter was 83%, compared to control cultures. Transcriptomic analysis revealed that the usage of green filters resulted in upregulation of transcripts involved in ribosome biogenesis or coding for elongation factors, exemplified by a 2.3-fold increase of TEF3. On the other hand, a 2.7-fold downregulation was observed in photosynthesis-related petJ. Exposure to blue filtered light led to the upregulation of transcripts associated with pyruvate metabolism, while photosynthesis was negatively impacted. In contrast, application of red filter induced minor transcriptomic alterations. Regarding metabolomic analysis, sugars, amino acids, and organic acids exhibited significant changes under different light regimes. For instance, under blue filtered light sucrose accumulated over 6-fold, while aspartic acid content decreased by 4.3-fold. Lipidomics analysis showed significant accumulation of heptadecanoic and linoleic acids under green and red light filters. Together, our findings indicate that filter light can be used for targeted metabolic manipulation.

Green synthesis and evaluation of dual herb-extracted DHM-AgNPs: Antimicrobial efficacy and low ecotoxicity in agricultural and aquatic systems

Uncontrolled applications of weedicide and fertilizer can harm the soil ecology, and most significantly, earthworms are hazardous soil engineers. Thus, we aimed at the toxicity and histopathological alterations in the earthworm Eudrilus euginae following exposure to glyphosate (weedicide), urea (fertilizer), and environmentally friendly dual herb-mixed silver nanoparticles (DHM-AgNPs). The DHM-AgNPs were synthesized using a blend of Alfinia officinarum and Curcuma longa aqueous leaf extracts with 1 mM silver nitrate. The color change from yellow to brown after an hour of incubation was a significant indicator of successful DHM-AgNP synthesis. Characterization of the DHM-AgNPs using UV–Vis spectra indicated a surface plasmon resonance (SPR) peak at 430 nm. In addition to FT-IR spectroscopy and XRD analysis, SEM, TEM, and SEM investigations were performed to identify the DHM-AgNPs. The XPS analysis revealed the oxidation state and surface chemical composition, and Ag NP's specific surface area and degree of porosity were measured using BET. Furthermore, different concentrations of urea and glyphosate were administered to Artemia nauplii and E. euginae to assess their toxicity. The mortality rate for E. euginae exposed to a higher urea concentration (10 g/kg of soil) was 100%. In contrast, a % mortality rate of 83% was noted at 0.5 g/kg of soil. The maximum mortality (90 ± 0.64%) was observed at a 10 mL/kg/L concentration for glyphosate. In contrast, low mortality was noted in E. euginae and A. nauplii exposed with gradient concentrations of DHM-AgNPs compared to glyphosate and urea. As aquaculture and foodborne diseases are widespread, DHM-AgNPs showed significant anti-Vibrio activity against pathogenic Vibrio-related bacteria, inhibiting 80% at 100,100 μg/L, which is of great concern. This study suggests the potential use of DHM-AgNPs in field aqua and crops culture for eco-friendly pest control and anti-Vibrio activity without causing soil and environmental pollution. Further research is warranted to determine the efficacy, safety, and cost-effectiveness of DHM-AgNPs in aqua and agricultural practices.

NMNAT1 Is Essential for Human iPS Cell Differentiation to the Retinal Lineage.

The gene encoding nicotinamide mononucleotide adenylyltransferase 1 (NMNAT1), a nicotinamide adenine dinucleotide synthetase localized in the cell nucleus, is a causative factor in Leber's congenital amaurosis, which is the earliest onset type of inherited retinal degeneration. We sought to investigate the roles of NMNAT1 in early retinal development. We used human induced pluripotent stem cells (hiPSCs) and established NMNAT1-knockout (KO) hiPSCs using CRISPR/cas9 technology to reveal the roles of NMNAT1 in human retinal development. NMNAT1 was not essential for the survival and proliferation of immature hiPSCs; therefore, we subjected NMNAT1-KO hiPSCs to retinal organoid (RO) differentiation culture. The expression levels of immature hiPSC-specific genes decreased in a similar manner after organoid culture initiation up to 2 weeks in the control and NMNAT1-KO. Neuroectoderm-specific genes were induced in the control and NMNAT1-KO organoids within a few days after starting the organoid culture; PAX6 and TUBB3 were higher in NMNAT1-KO organoids up to 7days than in the control organoids. However, the induction of genes involving retinal early development, such as RAX, which was induced at around day 10 in this culture, was considerably reduced in NMNAT1-KO organoids. Morphological examination also showed failure of retinal primordial structure formation, which became visible at around 2 weeks of the control culture, in the NMNAT1-KO organoids. Decreased intracellular NAD levels and poly(ADP-ribosyl)ation were observed in NMNAT1-KO organoids at 7 to 10days of the culture. Mass spectrometry analysis of inhibited proteins in the poly(ADP-ribosyl)ation pathway identified poly(ADP-ribosyl)ation of poly(ADP-ribose) polymerase 1 (PARP1) as a major protein. These results indicate that NMNAT1 was dispensable for neural lineage differentiation but essential for the commitment of retinal fate differentiation in hiPSCs. The NMNAT1-NAD-PARP1 axis may play a critical role in the appropriate development of human retinal lineage differentiation.