Days Of Incubation Research Articles

Inhibition of mTOR prevents glucotoxicity-mediated increase of SA-beta-gal, p16INK4a, and insulin hypersecretion, without restoring electrical features of mouse pancreatic islets.

An over-activation of the mechanistic target of rapamycin (mTOR) pathway promotes senescence and age-related diseases like type 2 diabetes. Besides, the regenerative potential of pancreatic islets deteriorates with aging. Nevertheless, the role of mTOR on senescence promoted by metabolic stress in islet cells as well as its relevance for electrophysiological aspects is not yet known. Here, we investigated whether parameters suggested to be indicative for senescence are induced in vitro in mouse islet cells by glucotoxicity and if mTOR inhibition plays a protective role against this. Islet cells exhibit a significant increase (~ 76%) in senescence-associated beta-galactosidase (SA-beta-gal) activity after exposure to glucotoxicity for 72h. Glucotoxicity does not markedly influence p16INK4a protein within 72h, but p16INK4a levels increase significantly after a 7-days incubation period. mTOR inhibition with a low rapamycin concentration (1nM) entirely prevents the glucotoxicity-mediated increase of SA-beta-gal and p16INK4a. At the functional level, reactive oxygen species, calcium homeostasis, and electrical activity are disturbed by glucotoxicity, and rapamycin fails to prevent this. In contrast, rapamycin significantly attenuates the insulin hypersecretion promoted by glucotoxicity by modifying the mRNA levels of Vamp2 and Snap25 genes, related to insulin exocytosis. Our data indicate an influence of glucotoxicity on pancreatic islet-cell senescence and a reduction of the senescence markers by mTOR inhibition, which is relevant to preserve the regenerative potential of the islets. Decreasing the influence of mTOR on islet cells exposed to glucotoxicity attenuates insulin hypersecretion, but is not sufficient to prevent electrophysiological disturbances, indicating the involvement of mTOR-independent mechanisms.

Polyethylenimine Inclusion to Develop Aqueous Alginate-Based Core-Shell Capsules for Biomedical Applications.

Aqueous core-shell structures can serve as an efficient approach that allows cells to generate 3D spheroids with in vivo-like cell-to-cell contacts. Here, a novel strategy for fabricating liquid-core-shell capsules is proposed by inverse gelation of alginate (ALG) and layer-by-layer (LbL) coating. We hypothesized that the unique properties of polyethylenimine (PEI) could be utilized to overcome the low structural stability and the limited cell recognition motifs of ALG. In the next step, alginate dialdehyde (ADA) enabled the Schiff-base reaction with free amine groups of PEI to reduce its possible toxic effects. Scanning electron microscopy and light microscopy images proved the formation of spherical hollow capsules with outer diameters of 3.0 ± 0.1 mm for ALG, 3.2 ± 0.1 mm for ALG/PEI, and 4.0 ± 0.2 mm for ALG/PEI/ADA capsules. The effective modulus increased by 3-fold and 5-fold when comparing ALG/PEI/ADA and ALG/PEI to ALG capsules, respectively. Moreover, PEI-coated capsules showed potential antibacterial properties against both Staphylococcus aureus and Escherichia coli, with an apparent inhibition zone. The cell viability results showed that all capsules were cytocompatible (above 75.5%). Cells could proliferate and form spheroids when encapsulated within the ALG/PEI/ADA capsules. Monitoring the spheroid thickness over 5 days of incubation indicated an increasing trend from 39.50 μm after 1 day to 66.86 μm after 5 days. The proposed encapsulation protocol represents a new in vitro platform for developing 3D cell cultivation and can be adapted to fulfill the requirements of various biomedical applications.

Sideritis raeseri—Modified Coatings on Ti-6Al-4V as a Carrier for Controlled Delivery Systems of Active Substances

The search for the ideal metallic material for an implant is still a difficult challenge for scientists due to the phenomenon of corrosion and the consequent disruption of the implant structure. Prevention is the application of coatings that protect the implant, activate the tissues for faster regeneration, and also prevent inflammation through antibacterial and antiviral effects. The present study focuses on the selection of components for a Ti-6Al-4V alloy coating. These days, researchers are taking an intense interest in extracts of natural origin. It was decided to take a look at Sideritis raeseri, which contains vitamins and valuable elements and is rich in polyphenols, as well as antioxidants. The composition of coatings based on a PEG polymer reinforced with brushite and the S. raeseri extract with the proteins L-carnosine, fibroin, or sericin was developed. The samples were subjected to detailed physiochemical analysis, including potentiometry and electrical conductivity analysis, Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), X-ray diffraction (XRD) analysis, and UV-VIS spectroscopy. The study demonstrated that polyphenols were successfully released from the coatings during incubation in vitro. The osteointegration process can be supported by a number of factors, such as the release of polyphenols from implant coatings to prevent bacterial, viral, and fungal infections. Subjecting the samples to 14 days of incubation demonstrated their interactions with the incubation fluids, an ion exchange between the medium and the materials. An analysis of the surface morphology exhibited the presence of brushite crystals and their increased number after incubation, indicating the bioactivity of the formed coatings.

Design of Highly Porous Materials Based on Chitosan/Pectin Interpolyelectrolyte Complex for Wound Healing Application

Interpolyelectrolyte complexes (IPECs) of polysaccharides are multifunctional polymer materials that improve the mechanical and physicochemical properties of individual polysaccharides. In this study, highly porous (>90%) materials based on IPECs of versatile natural polysaccharides, chitosan (30 and 1,200 kDa) and pectin, are obtained by freeze-drying technique. To enhance the interaction between chitosan and pectin macromolecules, the latter are chemically functionalized with dialdehyde groups. The chitosan-/aldehyde-functionalized pectin (Chit/AF-Pect) polyelectrolyte complex sponges obtained are characterized using SEM, FTIR spectroscopy, and TGA. The swelling capacity study reveals a higher swelling ratio of IPEC sponges with an increase in both the molecular weight and content of chitosan: for Chit30/AF-Pect, the swelling ratio rises from 327% to 480%, while for Chit1200/AF-Pect, from 681% to 1,066%. Additionally, the in vitro degradation test demonstrates higher stability of Chit1200/AF-Pect sponges in comparison with those of Chit30/AF-Pect: after 4 days of incubation, the weight losses are found to be 9%–16% and 18%–41%, respectively. The cytotoxicity study shows that Chit30/AF-Pect sponges are noncytotoxic, with cell viability values >70%. Furthermore, the Chit30/AF-Pect sponges, obtained at chitosan:pectin weight ratio of 5:1, exhibit bactericidal activity against Escherichia coli BIM B-984 G, Pseudomonas aeruginosa BIM B-807 G, Staphylococcus aureus BIM B-1841, and slightly inhibit the growth of Enterococcus faecalis BIM B-1530 G. These findings indicate that the obtained Chit30/AF-Pect sponges can be used to create wound dressings for wound healing applications.

Development of treatment strategies by comparing the minimum inhibitory concentrations and minimum fungicidal concentrations of azole drugs in dermatophytes.

We compared the minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) of azoles in antifungal drug-susceptible, terbinafine-resistant, and lowly itraconazole (ITCZ)-susceptible strains of dermatophytes. To assess the MICs of ITCZ, ravuconazole (RVCZ), efinaconazole (EFCZ), and luliconazole (LUCZ) in the isolates, broth microdilution assays were performed based on the Clinical and Laboratory Standards Institute M38-A2 guidelines with modifications. After the assays for determining the MICs, the inoculum suspensions in wells were resuspended, then 10 μL of the growth solution in each well was inoculated onto potato dextrose agar with the use of a pipette. After 7 days of incubation at 28°C, the MFCs were determined as the lowest concentration of a drug that allowed the growth of colonies on the potato dextrose agar. The MICs in the dermatophytes were <0.03 to >32 mg/L for ITCZ, <0.03 to 4 mg/L for RVCZ, <0.03 to 2 mg/L for EFCZ, and <0.03 mg/L for LUCZ. The MFCs in the dermatophytes were 1 to >32 mg/L for ITCZ, 0.06 to >32 mg/L for RVCZ, <0.03 to 4 mg/L for EFCZ, and <0.03 to 2 mg/L for LUCZ. If the drug susceptibility test shows that the fungi are resistant to the drug, the treatment can be changed to a susceptible drug in advance, or if the fungi are low-susceptible, the treatment can be done with the recognition that it may require a longer treatment period than usual.

Biodegradation of polyurethanes by Staphylococcus warneri and by microbial co-culture

With the increasing production and use of polyurethanes (PUs), it is necessary to develop sustainable techniques for the remediation of plastic pollution. The use of microorganisms capable of biodegrading PUs may be an environmentally desirable solution for controlling these plastic contaminants. To contribute to the discovery of alternatives for the mitigation of plastics in the environment, this study aimed to explore the potential of StaphylococcuswarneriUFV_01.21, isolated from the gut of Galleria mellonellalarvae, for biodegradation of PU in pure culture and microbial co-culture with Serratia liquefaciensL135. S. warneri grew using Impranil® PU as the sole carbon source in pure culture and co-culture. With six days of incubation, the biodegradation of Impranil® in Luria Bertani broth was 96, 88 and 76%, while in minimal medium, it was 58, 54 and 42% for S. warneri, S. liquefaciens, and co-culture, respectively. In addition, S. warneri in pure culture or co-culture was able to biodegrade, adhere and form biofilms on the surfaces of Impranil® disks and poly[4,4′-methylenebis (phenyl isocyanate)-alt-1,4-butanediol/di(propylene glycol)/polycaprolactone] (PCLMDI) films. Scanning electron microscopy also revealed biodegradation by detecting the formation of cracks, furrows, pores, and roughness on the surfaces of inoculated PU, both with pure culture and microbial co-culture. This study is the first to demonstrate the potential of S. warneriin PU biodegradation.

Microbe-cellulose hydrogels as a model system for particulate carbon degradation in soil aggregates

Abstract Particulate carbon (C) degradation in soils is a critical process in the global C cycle governing greenhouse gas fluxes and C storage. Millimeter-scale soil aggregates impose strong controls on particulate C degradation by inducing chemical gradients of e.g., oxygen, as well as limiting microbial mobility in pore structures. To date, experimental models of soil aggregates have incorporated porosity and chemical gradients but not particulate C. Here, we demonstrate a proof-of-concept encapsulating microbial cells and particulate C substrates in hydrogel matrices as a novel experimental model for soil aggregates. Ruminiclostridium cellulolyticum was co-encapsulated with cellulose in millimeter-scale polyethyleneglycol-dimethacrylate (PEGDMA) hydrogel beads. Microbial activity was delayed in hydrogel-encapsulated conditions, with cellulose degradation and fermentation activity being observed after 13 days of incubation. Unexpectedly, hydrogel encapsulation shifted product formation of R. cellulolyticum from an ethanol-lactate-acetate mixture to an acetate-dominated product profile. Fluorescence microscopy enabled simultaneous visualization of the PEGDMA matrix, cellulose particles, and individual cells in the matrix, demonstrating growth on cellulose particles during incubation. Together, these microbe-cellulose-PEGDMA hydrogels present a novel, reproducible experimental soil surrogate to connect single cells to process outcomes at the scale of soil aggregates and ecosystems.

An antifouling epoxy coated metal surface containing silica-immobilized carbonic anhydrase supraparticles for CO2 capture through microalgae

Carbonic anhydrase (CA) has a promising application as a green and efficient biocatalyst for CO2 capture, and many successful cases of immobilizing CA have been reported. However, CA antifouling coatings on metal for CO2 sequestration have rarely been reported. Herein, dimeric CA from Sulfurihydrogenibium azorense (SazCA) with a ferritin tag, which was prepared by low-speed centrifugation with high yield, was adopted as a free enzyme and encapsulated in the sol-gel silica. The silica-immobilized CAs were dispersed into the commercialized metal-antifouling epoxy resin paint to obtain CA coated nickel foams, which had excellent stability, with 90 % and 67 % residual activity after 28 days of incubation at 30 °C and 60 °C, respectively. The CA coated nickel foams remained 60 % original activity after 6 cycles of use within 28 days. Then, a CA-microalgae carbon capture device was constructed using the CA coated nickel foams and Chlorella. The growth rate of Chlorella was significantly increased and the biomass of Chlorella increased by 29 % compared with control after 7 days of incubation. Due to the simple and cost-effective preparation process, sustainable and efficient CO2 absorption, this easy-to-scale up CA coated nickel foam has great potential in CA assisted microalgae-based CO2 capture and carbon neutrality.

The effect of the 13C abundance of soil microbial DNA on identifying labelled fractions after ultracentrifugation

DNA-based stable isotope probing (DNA-SIP) technology has been widely employed to trace microbes assimilating target substrates. However, the fractions with labelled universal genes are sometimes difficult to distinguish when detected by quantitative real-time PCR. In this experiment, three paddy soils (AQ, CZ, and NB) were amended with 0.1% glucose containing 13C at six levels, and DNA was then extracted after a 7-day incubation and subjected to isopycnic gradient centrifugation. The results showed that the amount of labelled DNA was notably related to the 13C-glucose percentage, while the separation spans of 18S rRNA and 16S rRNA genes between labelled and unlabelled treatments became notably clearer when the δ13C values of the total DNA were 90.9, 61.6, and 38.9‰ and 256.2, 104.5 and 126.1‰ in the AQ, CZ, and NB soils, respectively. Moreover, fractionated DNA was also labelled by determining the δ13C values while adding only 5 atom% 13C-glucose to the soil. The results suggest that the optimal labelling fractions were not always those fractions with the maximal gene abundance, and detecting the δ13C values of the total and fractionated DNA was beneficial in estimating the results of DNA-SIP.Key points• Appropriate 13C-DNA amount was needed for DNA-SIP.• Detecting the 13C ratio of fractionated DNA directly was an assistant method for identifying the labelled fractions.• Fractions with the maximal 18S or 16S rRNA gene abundance always were not labelled.

First report of Colletotrichum queenslandicum associated with chilli anthracnose in Mauritius.

Globally, chilli (Capsicum annuum L.) is one of the most economically important and widely cultivated crop which elicits ethnomedicinal and nutritional potential as well as enhancing the taste and aroma of foods (Ayob et al., 2022; Kiran et al., 2020). Anthracnose disease is regarded as a prime constraint in chilli production, leading to enormous losses in tropical and subtropical countries. In September 2022, chilli fruit displaying sunken, shriveled and dark bown to black lesion with abundant acervuli on the surface was obtained from Flacq, Mauritius. From the symptomatic tissue, small pieces of the diseased tissue were excised, surface-disinfected using 1% sodium hypochlorite, twice rinsed using sterilized distilled water, air-dried and plated on PDA. After 7 days of incubation at room temperature, white to greyish white colony with dense white cottony aerial mycelium was recovered. Out of two isolates, CHF and CH10, the latter was considered for morphological and molecular characterization. The observed conidia (n=30) were unicellular, straight, cylindrical with rounded ends and slight constriction near the centre and had average length and width of 20.5 µm and 6 µm, respectively. For growth rate measurement of the isolate, two 5×5 mm of fungal agar plugs were taken from growing edge of colony, inoculated at centre of individual PDA plate and incubated at room temperature with a natural light/dark cycle. The diameter of the cultures were measured perpendicularly for a period of 7 days and the growth rate was calculated as 7-day average of daily growth (mm day-1). The growth rate of the fungal isolate (CH10) was 13.5 mm day-1 on PDA. Based on the morphological characters, the isolate was classified within the C. gloeosporioides species complex. For precise identification of the isolate, DNA was extracted from fungal mycelium using traditional DNA isolation methods (Ranghoo and Hyde, 2000), followed by PCR amplification and DNA sequencing using primer pairs ITS4/ITS5 (White et al., 1990), GDF/GDR and T1/Bt2b (Gan et al., 2016), respectively. ITS gene sequence (600 bp) confirmed that the isolate was Colletotrichum, with 99.83% similarity to KR704204 while GADPH (277 bp), TUB2 (733 bp) and ApMat (801 bp) gene sequences showed 99.64 to 100% similarity to C. queenslandicum with GenBank reference sequences, KT372374, KU221378 and MG674932 respectively. The gene sequences of isolate CH10 were deposited in GenBank database under the following accession numbers OR681557 (ITS), OR233734 (GADPH), OR475575 (TUB2) and PP622748 (ApMat). Koch's postulates were confirmed by spraying disease-free chilli plants with 10µL of conidial suspension (1 × 106 spores/ml) prepared from 7 days old colony of isolate CH10. Healthy chilli plants inoculated with sterile distilled water served as a negative control experiment. The plants were grown in pots in a moist chamber at 25˚C. After 5 days post-inoculation, anthracnose symptoms were developed on test plants while the control plant remained asymptomatic. The original isolate was successfully recovered from the test fruits, thus fulfilling Koch's postulates. The experiment was repeated thrice and revealed the same results. To the best of our knowledge, this is the first record of C. queenslandicum in Mauritius and is the first time to report anthracnose of chilli caused by this fungus. Colletotrichum queenslandicum has previously been reported in Europe, Mexico, US, Puerto Rico, Australia, Fiji, Brazil, Indonesia and China. Furthermore, the latter was associated with papaya, avocado, cashew, coffee, Persian lime, Licania tomentosa, white mangrove, lychee, mango, Nephelium lappaceum, olive, passionfruit, Dracaena cambodiana and Syzygium australe (Câmara and Vieira, 2022; Shidiq et al., 2024; Wang et al., 2022). This study will allow local farmers training and extension facilities to increase awareness among farmers about this disease-causing agent and allow them to take necessary measures for building up chilli crops resilience against this new and emerging pathogen in Mauritius.

Identification of compounds that cause axonal dieback without cytotoxicity in dorsal root ganglia explants and intervertebral disc cells with potential to treat pain via denervation.

Low back pain, knee osteoarthritis, and cancer patients suffer from chronic pain. Aberrant nerve growth into intervertebral disc, knee, and tumors, are common pathologies that lead to these chronic pain conditions. Axonal dieback induced by capsaicin (Caps) denervation has been FDA-approved to treat painful neuropathies and knee osteoarthritis but with short-term efficacy and discomfort. Herein, we propose to evaluate pyridoxine (Pyr), vincristine sulfate (Vcr) and ionomycin (Imy) as axonal dieback compounds for denervation with potential to alleviate pain. Previous literature suggests Pyr, Vcr, and Imy can cause undesired axonal degeneration, but no previous work has evaluated axonal dieback and cytotoxicity on adult rat dorsal root ganglia (DRG) explants. Thus, we performed axonal dieback screening using adult rat DRG explants in vitro with Caps as a positive control and assessed cytotoxicity. Imy inhibited axonal outgrowth and slowed axonal dieback, while Pyr and Vcr at high concentrations produced significant reduction in axon length and robust axonal dieback within three days. DRGs treated with Caps, Vcr, or Imy had increased DRG cytotoxicity compared to matched controls, but overall cytotoxicity was minimal and at least 88% lower compared to lysed DRGs. Pyr did not lead to any DRG cytotoxicity. Further, neither Pyr nor Vcr triggered intervertebral disc cell death or affected cellular metabolic activity after three days of incubation in vitro. Overall, our findings suggest Pyr and Vcr are not toxic to DRGs and intervertebral disc cells, and there is potential for repurposing these compounds for axonal dieback compounds to cause local denervation and alleviate pain.

Impact of lytic phages infecting Salmonella enterica subspecies arizonae on physicochemical and microbiological water quality of the old Port of Bizerte (Tunisia, SW Mediterranean Sea)

Phage therapy is among recent innovative technology to improve wastewater quality. Previous phage studies have focused on the interactions of phages with bacterial host-cells or the impact of environmental conditions on phage-host cell dynamics. In this research, we particularly examined the impact of phages on a marine physicochemical and microbiological environmental characteristic. Following three days of incubation, seawaters receiving separately phage fBC-Eco01 (OM272052) and phage PA25, undergone a temperature increase of 1.7°C and 2.5°C, respectively. Seawater salinity values saved an increase in conductivity over time that ranged between 51.03 and 53.94 µS and from 51.04 to 53.7 μS for the phage PA25 and the phage fBC-Eco01, respectively. A significant decrease in dissolved oxygen levels of 2,1mg/L were reported following the addition of both phages PA25 and fBC-Eco01 during three days of incubation, compared to a decrease of 1mg/L in the control seawaters. The TOC increased from 0.33 ppm to 0.67 ppm in the presence of the PA25 phage and from 0.2 ppm to 1.51 ppm after the addition of the fBC-Eco01 phage, compared to control that saved values decreasing from 0.35 ppm to 0.05 ppm. The absorbance and fluorescence spectrophotometry values were also affected by phages addition. Moreover, phages addition reduced total aerobic mesophilic bacteria (TAMB) and Total aerobic thermophilic bacteria (TATB) abundance, corresponding to a decrease of 2 Ulog and 1 Ulog discharge, respectively following addition of PA25 and 3 Ulog and 2 Ulog discharge, respectively with addition of fBC-Ec01 phage.

Impact of dichromatic lighted incubation on hatching result and post-hatch performance of broiler chickens.

This study was planned to evaluate the impact of dichromatic lights during incubation on the hatching and post-hatch performance of broiler chickens. A total of 500 eggs of broiler breeder (Ross 308; Age 44 weeks) were evenly divided according to a completely randomized design into 4 treatments having 5 replicates and 25 eggs each. Treatments consisted of dichromatic lights Blue + Red (BR), Green + Red (GR) and Green + Blue (GB) provided at an intensity of 250lx for 12h a day along with a Dark (D) environment. After hatching 200 chicks (50 from each respective light group) were divided into 4 treatments with 5 replicates each having 10 chicks. Results indicated a higher embryo index (13.12%) in the GR group on the 12th day of incubation; while an ideal hatch window was observed in GR and GB (98.18% and 96.00% hatched chicks) lighting groups. In hatching traits, higher hatchability (86.15) and hatch of fertile (93.85) percentages were observed in GR lighting followed by GB, BR and Dark treatment groups; while dead-in shell embryos were lowest in the GR group. In growth performance, higher feed intake (513.20g) and body weight (479.20g) were observed in the GB group followed by GR, BR and dark group; and feed conversion ratio (FCR) was better in the GR group (1.06). In welfare parameters, improved physical asymmetry (0.90mm) and tonic immobility (54.40s) were measured in the GR group followed by GB, BR and the dark group. It was concluded that under experimental conditions when broiler breeder eggs are provided with GR lighting during incubation, it can help to improve hatchability, growth performance and welfare traits in chicks.

In vitro inhibition of Fusarium spp. isolated from bean crop using Argemone ochroleuca extract

Species of the genus Fusarium are the main cause of root rot in bean crops, resulting in yield losses of up to 50%. Synthetic fungicides are the primary tools for their control; however, they cause environmental damage. The objective of this study was to evaluate the effect of extracts from A. ochroleuca on Fusarium spp. under in vitro conditions. During the spring-summer 2022 growing season, plants exhibiting symptoms of wilting and vascular browning were collected. The isolated fungi were purified through monosporic cultures and hyphal tip techniques, and their morphological characteristics were characterized. To determine growth inhibition, a poisoned medium methodology was employed, evaluating two concentrations of extracts (13% and 23%) in both ethanol and aqueous mediums. The obtained isolates corresponded to the morphological characteristics of Fusarium. The extract volumes that showed 100% inhibition were the 23% concentration in both ethanol and aqueous mediums. The 13% ethanol extract exhibited inhibition ranging from 29% to 62% between the first and tenth day of incubation. The 13% aqueous extract displayed inhibition ranging from 25% to 43%. The Argemone ochroleuca extract demonstrates antifungal properties against Fusarium spp. isolated from beans.

Iron-based materials functionalized with carbon and phosphorus recovered from sludge enhanced the formation of stable minerals to passivate lead and chromium in wastewater and soil

The bioaccumulation and toxicity of heavy metals are serious threats to human activities and ecological health. The exploitation of environmentally friendly passivated materials is major importance for the remediation of heavy metal contaminated soil. This research developed a new type of environmental functional material with a core-shell structure, which is an iron-based material functionalized with phosphorus and carbon from sludge for heavy metal pollution remediation. The results indicated that the C/P@Fe exhibits excellent heavy metal removal ability, and the maximum removal rates of the two heavy metals in simulated wastewater could reach 100% under optimum reaction conditions. It also effectively converts the labile Cr/Pb into the stable fraction after 28 days of incubation, which increased the maximum residual fraction percentage of Cr and Pb by 32.43% and 160% in soil. Further analysis found that the carbon layer wrapped around the iron base could improve the electron transport efficiency of reducing iron, phosphorus and ferrum could react with heavy metal ions to form stable minerals, such as FeCr2O4, FeO·Cr2O3, Pb5(PO4)3OH, PbCO3, 2PbCO3·Pb(OH)2 and PbS, after reacting with C/P@Fe. The study demonstrated that the Iron-based materials functionalized with carbon and phosphorus from sludge provided a more efficient way to remove heavy metals.

Novel polyurethane-degrading cutinase BaCut1 from Blastobotrys sp. G-9 with potential role in plastic bio-recycling

Environmental pollution caused by plastic waste has become global problem that needs to be considered urgently. In the pursuit of a circular plastic economy, biodegradation provides an attractive strategy for managing plastic wastes, whereas effective plastic-degrading microbes and enzymes are required. In this study, we report that Blastobotrys sp. G-9 isolated from discarded plastic in landfills is capable of depolymerizing polyurethanes (PU) and poly (butylene adipate-co-terephthalate) (PBAT). Strain G-9 degrades up to 60% of PU foam after 21 days of incubation at 28 ℃ by breaking down carbonyl groups via secretory hydrolase as confirmed by structural characterization of plastics and degradation products identification. Within the supernatant of strain G-9, we identify a novel cutinase BaCut1, belonging to the esterase family, that can reproduce the same effect. BaCut1 demonstrates efficient degradation toward commercial polyester plastics PU foam (0.5 mg enzyme/25 mg plastic) and agricultural film PBAT (0.5 mg enzyme/10 mg plastic) with 50% and 18% weight loss at 37 ℃ for 48 h, respectively. BaCut1 hydrolyzes PU into adipic acid as a major end-product with 42.9% recovery via ester bond cleavage, and visible biodegradation is also identified from PBAT, which is a beneficial feature for future recycling economy. Molecular docking, along with products distribution, elucidates a special substrate-binding modes of BaCut1 with plastic substrate analogue. BaCut1-mediated polyester plastic degradation offers an alternative approach for managing PU plastic wastes through possible bio-recycling.

Evaluation of the Mutagenic Potential of Artavol® Using the Ame’s Test

Background: The worldwide increase in the use of herbal medicinal - products calls for their safety testing to protect the public from unintended hazardous effects. However, most are not tested, putting the public at risk. The modified Ames ISO test is a useful resource for determining the mutagenic potential of medicinal products and was used in this study to determine the safety of an herbal tea like Artavol®. Methods: This study used Ame's Modified ISO test to analyze aqueous extract of Artavol®, in a Level II biosafety cabinet. Artavol® was extracted using the infusion method 1,2, freeze dried, diluted to concentrations of 125µg, 250µg, and 500µg and used in the study. Mutagenicity was tested by culturing Salmonella typhimurium TA98 and TA100. Results were considered by observing colour change in the wells of the microtitre plates from purple to yellow indicating mutation. Valid results were determined by comparing negative and positive control plates after 3 days of incubation, with positive results showing a color change from purple to yellow. Results: Colour changes were not observed in all wells containing 125µg, 250µg, and 500µg of Artavol® up to day 3, while negative control showed color changes equivalent to 80% for TA98 and 70% for TA100. Conclusion: Aqueous extract of Artavol® is not mutagenic. Further safety test such as sub-chronic toxicity study ad teratogenicity studies are recommended to provide more safety data on the product.

Formation and development of lamellar histoarchitecture of the ventricular myocardium of chick embryos

Background. The existence of several models of the myolamellar structure of the ventricular myocardium, which currently has a number of contradictory provisions, reflects the need for a reasonable integration of the results of different methods. Under these circumstances, the study of those ontogenetic mechanisms that are responsible for the formation and development of the myolamellar architecture of the myocardium is of great interest. The purpose of the study is to determine the ontogenetic transformations of the embryonic chicken heart that ensure the formation and development of the myolamellar structure of the ventricular myocardium. Methods. The work examined the embryos of Cobb500 cross chickens from the beginning of the 10th day to the 21st day of incubation. The lamellar organization of the ventricular myocardium was studied using light and transmission electron microscopy. Results. Starting from the 36th stage according to NN (the beginning of the 10th day of incubation), the active development of the stromal component was observed in the heart of chicken embryos, which led to the division of the tissue of the compact ventricular myocardium into groups of muscle fibers in the form of narrow elongated flat plates containing thicker than 3 to 5 rows of cardiomyocytes. At the 41st and 43rd stages of development, active development of the intercellular matrix and division of the myocardium mass into muscle plates continued as part of the compact ventricular myocardium. The intercellular spaces within the plates narrowed, and between the myolamellae, the perimysium accumulated elements of the microcirculatory bed, functionally active fibroblasts, a large amount of amorphous substance, and bundles of formed collagen fibers. At the final stages of embryogenesis, the muscle plates of the left ventricle acquired a pronounced spiral orientation with a gradual displacement of the long axis of the muscle fibers in the direction from the apex of the ventricle to its base. In the wall of the right ventricle, the location of the myolamella acquired a transverse oblique-circular orientation. Conclusion. A comparison of the structure and geometry of the myolamella made it possible to reveal that starting from the 38th stage of development in the left ventricle, the conditions for the translational-rotational mechanism of chamber contraction were formed and increased, in which the formation of the difference between the systolic and diastolic volumes of the left ventricle is ensured not only by the longitudinal apico-basal vector compression of the cavity, but also by mutual sliding of spirally oriented plates in the ventricular wall. In right ventricle, the contraction mechanism is based on the longitudinal-circular compression of the chamber in accordance with the oblique-circular orientation of the muscle fibers in the composition of the myolamella without displacement in the state of systole.

Fungal Bioremediation of the β-Lactam Antibiotic Ampicillin under Laccase-Induced Conditions

Due to widespread overuse, pharmaceutical compounds, such as antibiotics, are becoming increasingly prevalent in greater concentrations in aquatic ecosystems. In this study, we investigated the capacity of the white-rot fungus, Coriolopsis gallica (a high-laccase-producing fungus), to biodegrade ampicillin under different cultivation conditions. The biodegradation of the antibiotic was confirmed using high-performance liquid chromatography, and its antibacterial activity was evaluated using the bacterial growth inhibition agar well diffusion method, with Escherichia coli as an ampicillin-sensitive test strain. C. gallica successfully eliminated ampicillin (50 mg L−1) after 6 days of incubation in a liquid medium. The best results were achieved with a 9-day-old fungal culture, which treated a high concentration (500 mg L−1) of ampicillin within 3 days. This higher antibiotic removal rate was concomitant with the maximum laccase production in the culture supernatant. Meanwhile, four consecutive doses of 500 mg L−1 of ampicillin were removed by the same fungal culture within 24 days. After that, the fungus failed to remove the antibiotic. The measurement of the ligninolytic enzyme activity showed that C. gallica laccase might participate in the bioremediation of ampicillin.

Assessing the influence of micelle silymarin on laying hens' performance, egg quality, water loss and blood profile.

Micelle silymarin (MS) is known for its various beneficial properties, including antiswelling, antioxidant, hepatoprotective and antidiabetic effects. The primary goal of this research was to investigate how MS impacts the performance, egg quality, water loss and blood profile of laying hens. 288 Hy-Line brown laying hens, 28 weeks in age, were utilized for this experiment. The hens were randomly allocated into three dietary treatment groups, with each group comprising eight replicates of 12 hens, each housed in individual pens with access to feed and water. Over a 12-week feeding trial, the hens were provided with a basal diet supplemented with different levels of MS: 0%, 0.03% and 0.06%. The results indicated that the inclusion of MS in the hens' diet did not have a significant impact on their performance (p > 0.05). However, Haugh units, egg weight and eggshell strength showed a linear improvement (p < 0.05) throughout the entire trial period with MS supplementation. Furthermore, there was a linear decrease in egg yolk colour and eggshell thickness showed linear improvements (p < 0.05), particularly during Week 8, with MS supplementation. Moreover, layers fed diet supplemented with MS showed a linear increased (p < 0.05) in albumen height and eggshell thickness in Week 12. In addition, egg water loss during Week 12, the third day of incubation, linearly decreased as an effect of the increasing level of the MS in the laying hen's diet (p < 0.05). Regarding blood profile parameters, the study revealed a tendency for alkaline phosphatase to decrease, whereas aspartate aminotransferase and cholesterol levels were linearly decreased (p < 0.05). In summary, increasing the level of MS supplementation in the diet of laying hens appeared to be beneficial in improving egg quality, slight improvement for egg water loss and certain aspects of blood profile parameters, without adversely affecting the hens' growth performance.