Effect of ß-Alanine Metabolite on Gut Integrity and Immunity in Commercial Broiler Chickens Infected with Eimeria maxima.

(1) Background: In a metabolomics analysis conducted to investigate the mechanisms behind the growth-promoting effects of probiotics in broilers, ß-alanine was found to be significantly elevated. This led to the hypothesis that ß-alanine could also contribute to growth-promoting effects in infected broilers. (2) Methods: An in vitro culture system was developed to assess ß-alanine's impact on proinflammatory cytokine response in chicken macrophage cells, gut integrity in chicken intestinal epithelial cells, and muscle differentiation in quail muscle cells and primary chicken embryonic muscle cells. In vivo animal feeding studies were then conducted to investigate the effects of dietary ß-alanine on various disease parameters in Eimeria maxima-infected broiler chickens. (3) Results: In vitro, ß-alanine treatment significantly decreased the gene expression of cytokines in chicken macrophage cells and increased occuldin expression in chicken intestinal epithelial cells. Dietary ß-alanine increased the body weight of chickens following Eimeria maxima infection in the H-ALA group. Dietary ß-alanine also suppressed cytokines and increased JAM-2 and occludin expression in the H-ALA group compared to the infected group without ß-alanine supplementation. (4) Conclusions: These results strongly support the positive effects of dietary ß-alanine on intestinal immune responses and gut barrier function in broiler chickens infected with Eimeria maxima.

The Role of the Basophil Activation Test in the Diagnosis of Drug-Induced Anaphylaxis.

The diagnosis of drug-induced anaphylaxis (DIA) is a serious health problem. The Basophil activation test (BAT) is considered a specific in vitro provocation, and compared to in vivo provocation, it is more convenient, cheaper, and safer for the patient. This study aimed to evaluate the usefulness of the BAT in the diagnosis of DIA. This study included 150 patients referred to a reference allergy clinic with suspected drug allergies. All patients underwent a detailed clinical evaluation supplemented with the BAT. Positive BAT results were obtained in two out of 21 patients who were to receive the COVID-19 vaccine. The sensitivity and specificity of the BAT were 40% and 75% for the COVID-19 vaccine, 67% and 58% for DMG PEG 2000, and 100% and 75% for PEG 4000, respectively. Nine out of 34 patients with suspected antibiotic allergies had positive BAT results with 14 different antibiotics. Positive BAT results were also obtained with NSAIDs in two patients and with local anesthetics in three patients. The confirmation of allergy by the BAT improves the safety profile of the diagnostic work-up as it may defer the need for drug provocation, preventing potential anaphylactic reactions.

Synthesis, Characterization, and Biological Performances of Magnesium-Substituted Dicalcium Phosphate Anhydrous.

Dicalcium phosphate anhydrous (DCPA, CaHPO4) is regarded as an orthopedic material due to its ability to match the generation of new bone to the rate of implant resorption without considering the material's mechanical stability. Additionally, magnesium (Mg) is widely recognized for its essential function in bone metabolism, especially during the initial phases of osteogenesis. Therefore, we explored the influences of Mg ions on DCPA powder, in biological responses, and on the enhancement of osteogenic properties. Mg-DCPA powders with varying substitution levels (0, 3, 5, and 7 mol%) were produced using the co-precipitation method. In the in vitro test, precipitates began to develop on the surface of the Mg-DCPA powders after 7 days. These results indicate that Mg ions in the DCPA powder could enhance the generation of a new apatite phase when subjected to physiological fluids on the surface of the powder. In addition, the osteogenic performance of the DCPA powder was improved by adding Mg ions. The most effective magnesium substitution content in the DCPA powder in order to improve its osteogenic potential was approximately 3 mol%. Consequently, this amount of magnesium in the DCPA powder could control the maintaining time in the implantation operation to produce a new apatite phase.

Identification of novel RANKL inhibitors through in silico analysis.

Receptor activator of nuclear factor-κB ligand (RANKL) is considered the principal regulator of osteoclast differentiation. Therefore, strategies interfering with the RANKL-RANK signaling pathway may effectively inhibit osteoclast differentiation and mitigate bone resorption. Consequently, RANKL has become a promising target for new drug design strategies. Despite extensive research on specific drugs and antibodies, only a few have shown efficacy in treating osteoporosis. To address this challenge, we aimed to explore new approaches for designing drugs for osteoporosis. In this study, a 3D quantitative structure-activity relationship (QSAR) pharmacophore model was built for RANKL with reference to known inhibitor IC50 values. The optimal pharmacophore model was then employed as a 3D query to screen databases for novel lead compounds. The obtained compounds were subjected to ADMET and TOPKAT analyses to predict drug pharmacokinetics and toxicity. Molecular docking and de novo evolution approaches were applied to verify the docking binding affinities of the compounds. Five candidate compounds were subjected to further in vitro analyses to assess their anti-osteoporotic effects, among which compound 4 demonstrated significant inhibitory activity, achieving an inhibitory rate of 92.6 % on osteoclastogenesis at a concentration of 10 µM. Subsequent molecular dynamics (MD) simulations to assess the stability and behavior of compound 4 and its evolved variant, ZINC00059014397_Evo, within the RANKL binding site revealed that the variant is a potential therapeutic agent for targeting osteoclasts. This study offers valuable insights for developing next generation RANKL inhibitors for osteoporosis treatments.

Exploring the antiparasitic and antimicrobial potential of Schinus terebinthifolius Raddi essential oil against fish and shrimp pathogens.

This study aimed to perform in vitro antiparasitic and antimicrobial tests with the essential oil (EO) of Schinus terebinthifolius against of fish and shrimp. The chemical composition of the EO of S. terebinthifolius was determined by gas chromatography. For the antiparasitic test, the protozoan Epistylis sp. obtained from parasitized Oreochromis niloticus was used, and exposed to different concentrations of EO (2%, 1%, 0.5%, 0.25%), and control with 1% grain alcohol. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) test with EO of S. terebinthifolius evaluated the antimicrobial potential, with serial dilutions starting at 2% and control with 1% grain alcohol, using the strains of Aeromonas hydrophila (2.2 × 108 CFU mL-1), Edwardsiella tarda, Vibrio parahaemolyticus, V. harveyi, and V. alginolyticus (2.0 × 108 CFU mL-1). Chemical analysis revealed that the major EO compounds of S. terebinthifolius were δ-3-Carene (56.00%) and α-Pinene (16.89%). In the antiparasitic test, the concentration of 2% EO showed 100% efficacy against Epistylis sp. within 5 min. In the antimicrobial tests, the concentration of 2% EO was effective against all bacteria tested. The EO of S. terebinthifolius demonstrated antiparasitic and antimicrobial activity at a concentration of 2%, standing out as an alternative to conventional antibiotics.

Polyethylene Wear Testing of a Non-Mechanically Linked Total Elbow Replacement.

BACKGROUND: The objective of this study was to perform a polyethylene wear test on a non-mechanically linked total elbow arthroplasty implant using a clinically relevant in-vitro elbow wear test methodology that simulated ten years of use in the light to moderate activity of daily living range. MATERIALS AND METHODS: The test protocol applied an 80° arc of ulnohumeral motion beginning at 30° shy of full extension and progressing to 110° of flexion. Force was applied at 7° to recreate a valgus load on the elbow. A variable joint load profile at a frequency of 0.5 Hz was employed. The implants were tested for 5 million cycles (Mc) in a bovine serum lubricant. Implant component failure was characterized and polyethylene wear was determined gravimetrically. RESULTS: After 5 Mc the small polyethylene bushing wear rate was 0.56 mg/Mc. The medium size wear rate was 0.28 mg/Mc. Three large sizes were tested and the average wear rate was 0.39 ± 0.07 mg/Mc. No implant failure was identified. CONCLUSION: The test recreated an in vivo loading environment and measured polyethylene wear rates at specified cycle counts. The test demonstrated less wear than other joint replacements. Further clinical evaluation is necessary to determine if this translates into reduced complications of total elbow replacement associated with wear.

The effect of gut microbiota-derived carnosine on mucosal integrity and immunity in broiler chickens challenged with Eimeria maxima.

In the first study, an in vitro culture system was developed to investigate the effects of carnosine on macrophage proinflammatory cytokine response using an established chicken macrophage cell line (CMC), gut integrity using a chicken intestinal epithelial cell line (IEC), muscle differentiation in quail muscle cells (QMCs) and primary chicken embryonic muscle cells (PMCs), and direct anti-parasitic effect against Eimeria maxima sporozoites. Cells to be tested were seeded in 24-well plates and treated with carnosine at 4 different concentrations (0.1, 1.0, and 10.0 µg). After 18 h of incubation, cells were harvested to measure gene expression of proinflammatory cytokines in CMC, tight junction (TJ) proteins in IECs, and muscle cell growth markers in QMCs and PMCs. In vivo trials were conducted to investigate the effect of dietary carnosine on disease parameters in broiler chickens challenged with E. maxima. One hundred and twenty male broiler chickens (0-day-old) were allocated into 4 treatment groups: 1) basal diet without infection (NC), 2) basal diet with E. maxima infection (PC), 3) carnosine at 10.0 mg/kg feed with PC (HCS), and 4) carnosine at 1.0 mg/kg feed with PC (LCS). All groups except NC were orally infected with E. maxima on d 14. Jejunal samples were collected for lesion scoring and jejunum gut tissues were used for transcriptomic analysis of cytokines and TJ proteins. In vitro, carnosine treatment significantly decreased IL-1ß gene expression in CMC following LPS stimulation. In vivo feeding studies showed that dietary carnosine increased BW and ADG of chickens in E. maxima-infected groups and reduced the jejunal lesion score and fecal oocyst shedding in HCS group. Jejunal IL-1ß, IL-8, and IFN-γ expression were suppressed in the HCS group compared to PC. The expression levels of claudin-1 and occludin in IECs were also increased in HCS following carnosine treatment. In conclusion, these findings highlight the beneficial effects of dietary carnosine supplementation on intestinal immune responses and gut barrier function in broiler chickens exposed to E. maxima infection.

Effect of cassava pulp treated with Lactobacillus casei TH14, urea, and molasses on gas kinetics, rumen fermentation, and degradability using the in vitro gas technique.

This study focused on examining the gas dynamics, rumen fermentation, and digestibility of ensiled cassava pulp (CSVP) using Lactobacillus casei TH14, urea, and molasses in the context of a laboratory experiment. All data in this study were analyzed using treatments arranged in 2 × 2 × 2 factorial arrangements using a completely randomized design. The L.casei TH14 additive (L) was factor A. Factor B was the molasses additive (M), while factor C was urea (U). There was no interaction effect of L, U, and M on gas production, volatile fatty acid (VFA) content, pH value, or ammonia-nitrogen level (P<0.05). The interaction of L, U, and M influenced in vitro dry matter digestibility (IVDMD) at 12 h (P < 0.05), and the CSVP fermented with the additions of L, U, and M together (LUM) was higher than the additions of CON, M, U, UM, and L on IVDMD (P < 0.05). However, the IVDMD values of adding LUM were higher in the control group (CON), M, U, UM, and L additive groups (P < 0.05). There was an interaction effect of L, U, and M on the protozoal count at 8 h (P<0.05), which had a lower protozoal count in the control group. In addition, acetic acid and butyric acid concentrations at 4 h and 8 h (P<0.05) were increased during the fermentation of CSVP using L and M combinations. Furthermore, the combination of U and M enhanced (P<0.05) average acetic acid, propionic acid, and pH at 4 h and 8 h while reducing (P<0.05) the gas generation from the insoluble portion (b). It was suggested that utilizing L. casei TH14 together with urea and molasses can enhance nutrient contents and improve the in vitro dry matter digestibility of CSVP, although it has no effect on ruminal fermentation or gas production.

In vitro efficacy of essential oils against various Candida species.

Candida species are responsible for the most common fungal infections worldwide. We studied the in vitro antifungal activity of a large panel of essential oils (EOs) against various Candida species. The EOs activity against Candida spp. was tested using a gradient microdilution assay ranging from 4% to 0.008% (v/v). After a preliminary screening including 31 EOs, seven selected EOs were tested against 13 clinical isolates and four reference strains belonging to six Candida species. Cinnamomum zeylanicum and Cymbopogon giganteus EOs exhibited the best antifungal activity against all clinical and reference strains, with MIC ranges of 0.015%-0.25% (v/v). EOs from Litsea citrata, Backhousia citriodora and Ocimum sanctum presented MIC ranges of 0.03%-0.5% (v/v). The antifungal efficacy of EOs was independent of the susceptibility of Candida strains to usual antifungal agents. These EOs could have a promising antifungal action.

A Human Skin Explant Test as a Novel In Vitro Assay for the Detection of Skin Sensitization to Aggregated Monoclonal Antibodies.

Introduction: Monoclonal antibodies (mAbs) are important therapeutics. However, the enhanced potential for aggregation has become a critical quality parameter during the production of mAbs. Furthermore, mAb aggregation may also present a potential health risk in a clinical setting during the administration of mAb therapeutics to patients. While the extent of immunotoxicity in patient populations is uncertain, reports show it can lead to immune responses via cell activation and cytokine release. In this study, an autologous in vitro skin test designed to predict adverse immune events, including skin sensitization, was used as a novel assay for the assessment of immunotoxicity caused by mAb aggregation. Material and Methods: Aggregation of mAbs was induced by a heat stress protocol, followed by characterization of protein content by analytical ultra-centrifugation and transmission electron microscopy, revealing a 4% aggregation level of total protein content. Immunotoxicity and potential skin sensitization caused by the aggregates, were then tested in a skin explant assay. Results: Aggregated Herceptin and Rituximab caused skin sensitization, as shown by histopathological damage (grade II-III positive response) together with positive staining for Heat Shock Protein 70 (HSP70). Changes in T cell proliferation were not observed. Cytokine analysis revealed a significant increase of IL-10 for the most extreme condition of aggregation (65 °C at pH3) and a trend for an overall increase of IFN-γ, especially in response to Rituximab. Conclusions: The skin explant assay demonstrated that aggregated mAbs showed adverse immune reactions, as demonstrated as skin sensitization, with histopathological grades II-III. The assay may, therefore, be a novel tool for assessing immunotoxicity and skin sensitization caused by mAb aggregation.

The Accomplishments of KoCVAM in the Development and Implementation of Alternative Methods in Korea.

The Korean Center for the Validation of Alternative Methods (KoCVAM), which promotes the Three Rs principles and the use of alternative methods in Korea, has been operating within the Toxicological Screening and Testing Division of the Ministry of Food and Drug Safety (MFDS) since 2009. KoCVAM has exchanged opinions and information on the development and validation of non-animal alternative test methods as part of the International Cooperation on Alternative Test Methods (ICATM), and provided input into draft OECD Test Guidelines (TGs). Several Korean laws (e.g. the Cosmetics Act) encourage the use of alternative test methods for chemical testing and assessment. To promote and support the use of alternative test methods in the country, KoCVAM has published information and provided training on the national guidelines, which are based on the OECD TGs. In addition, KoCVAM has held annual training workshops on alternative test methods, to help Korean research institutions (including GLP test facilities) to implement them. In addition, by helping to develop and validate alternative test methods that were adopted in OECD TG 442B, TG 492 and TG 439, KoCVAM has contributed to the enhanced competitiveness of Korean industry on the worldwide stage.

Effect of decentration and tilt on the in vitro optical quality of monofocal and trifocal intraocular lenses.

PURPOSE: To evaluate and compare the effect of decentration and tilt on the optical quality of monofocal and trifocal intraocular lenses (IOL). METHODS: Optical quality of a monofocal IOL (AcrySof IQ SN60WF; Alcon Laboratories, Inc., USA) and a trifocal IOL (AcrySof IQ PanOptix; Alcon Laboratories, Inc., USA) was assessed using an in vitro optical bench (OptiSpheric IOL R&D; Trioptics GmbH, Germany). At apertures of 3.0 mm and 4.5 mm, modulation transfer function (MTF) at spatial frequency of 50 lp/mm, MTF curve and the United States Air Force (USAF) resolution test chart of the two IOLs were measured and compared at their focus with different degrees of decentration and tilt. Optical quality at infinity, 60 cm and 40 cm and the through-focus MTF curves were compared when the two IOLs were centered at apertures of 3.0 mm and 4.5 mm. Spectral transmittance of the two IOLs was measured by the UV-visible spectrophotometer (UV 3300 PC; MAPADA, China). RESULTS: The SN60WF and the PanOptix filtered blue light from 400 to 500 nm. Both IOLs at the far focus and the PanOptix at the intermediate focus showed a decrease in optical quality with increasing decentration and tilt. The PanOptix demonstrated enhanced optical quality compared to the previous gradient at the near focus at a decentration range of 0.3-0.7 mm with a 3.0 mm aperture, and 0.5 mm with a 4.5 mm aperture, whereas other conditions exhibited diminished optical quality with increasing decentration and tilt at the focus of both IOLs. When the two IOLs were centered, the SN60WF had better optical quality at infinity, while the PanOptix had better optical quality at 60 cm and 40 cm defocus. The optical quality of the SN60WF exceeded that of the PanOptix at far focus, with a 3 mm aperture decentration up to 0.7 mm and a 4.5 mm aperture decentration up to 0.3 mm; this observation held true for all tilts, irrespective of aperture size. As both decentration and tilt increased, the optical quality of the SN60WF deteriorated more rapidly than that of the PanOptix at the far focal point. CONCLUSIONS: The SN60WF showed a decrease in optical quality with increasing decentration and tilt. Optical quality of the PanOptix at the near focus increased in some decentration conditions and decreased in some conditions, while it showed a decrease at the other focuses with increasing decentration. While tilt only had a negative effect on optical quality. When both IOLs were centered, the PanOptix provided a wider range of vision, while the SN60WF provided better far distance vision. At the far focus, the SN60WF has better resistance to tilt than the PanOptix, but the optical quality degrades more quickly when decentered and tilted.

Development of natural cosmetic emulsion using the by-product of Lecythis pisonis seed.

Natural products and their biological activities are currently a subject of great interest to the industrial and scientific sector, due to society's awareness of the proper use of biodiversity and economic and sustainability. To promote the sustainable use of biomass the extract of the by-product of the shell seed of Lecythis pisonis was applied to develop a natural cosmetic emulsion. To ensure safety for its topical use the cytotoxic activity of its crude extract was evaluated by the colorimetric method of 3- bromide (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium, MTT, in cell culture of fibroblasts L929, human keratinocytes HaCat, and human endothelium EA.hy926 cell lines. The triplicate of the cosmetic formulation containing the extract was obtained regarding stability according to the procedures of the Brazilian Health Regulatory Agency (Anvisa). The irritability tests were performed using alternative methods, in vitro, chorioallantoic membrane assay (HET-CAM and CAM-TBS), and hemolysis test (RBC). The crude extract was not cytotoxic, IC50 index >780 mg/mL. The preservative system was effective against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and Aspergillus niger. The emulsion was classified as non-irritable. The crude extract of the by-product of sapucaia seeds can be incorporated into a natural emulsion, safe for topical use.

The larvicidal effect of the supernatant of Lactobacillus acidophilus ATCC 4356 on Toxocara canis.

Human toxocariasis is a parasitic anthropozoonosis that is difficult to treat and control. A previous study carried out with Lactobacillus acidophilus ATCC 4356 revealed that the cell free supernatant (CFS) of this probiotic killed 100% of Toxocara canis larvae in vitro. The present study aimed to investigate the characteristics of the CFS of L. acidophilus ATCC 4356, which may be involved in its larvicidal effects on T. canis. L. acidophilus ATCC 4356 was cultured, and lactic and acetic acids present in the CFS were quantified by high performance liquid chromatography (HPLC). The levels of pH and H2O2 were also analyzed. To assess the larvicidal effect of the CFS, this was tested pure and diluted (1:2 to 1:128) on T. canis larvae. High concentrations of lactic and acetic acids were detected in the CFS. The acidity of the pure CFS was observed at pH 3.8, remaining acidic at dilutions of 1:2 to 1:16. Regarding the in vitro larvicidal effect, 100% death of T. canis larvae was observed using the pure CFS and 1:2 dilution. Based on these results, it can be inferred that the presence of higher concentrations of organic acids and low pH of the medium contributed to the larvicidal activity of the CFS of L. acidophilus ATCC 4356. In addition, the maintenance of the larvicidal effect, even after dilution, suggests a greater chance of the larvicidal effect of this CFS against T. canis in vivo.

Influences of coexisting aged polystyrene microplastics on the ecological and health risks of cadmium in soils: A leachability and oral bioaccessibility based study.

Whether coexisting microplastics (MPs) affect the ecological and health risks of cadmium (Cd) in soils is a cutting-edge scientific issue. In this study, four typical Chinese soils were prepared as artificially Cd-contaminated soils with/without aged polystyrene (PS). TCLP and in vitro PBET model were used to determine the leachability (ecological risk) and oral bioaccessibility (human health risk) of soil Cd. The mechanisms by which MPs influence soil Cd were discussed from direct and indirect perspectives. Results showed that there was no significant difference in the leachability of soil Cd with/without aged PS. Additionally, aged PS led to a significant decrease in the bioaccessibility of soil Cd in gastric phase, but not in small intestinal phase. The increase in surface roughness and the new characteristic peaks (e.g., Si-O-Si) of aged PS directly accounted for the change in Cd bioaccessibility. The change in organic matter content indirectly accounted for the exceptional increase in Cd bioaccessibility of black soil with aged PS in small intestinal phase. Furthermore, the changes in cation exchange capacity and Cd mobility factor caused by aged PS explained the change in Cd leachability. These results contribute to a deeper understanding about environmental and public health in complicated emerging scenarios.

Endotoxin test by recombinant Factor C for 0.9% sodium chloride injection

Endotoxin contamination is a threat to the safety of pharmaceutical products, especially parenteral drugs. Any sterile and/or pyrogen-free pharmaceutical product requires regulatory specifications to ensure safe patient use. This study covers the performance evaluation study of an endotoxin quantitation commercial kit by recombinant Factor C (rFC), Endozyme II® Go, for 0.9% sodium chloride injection. The samples were spiked with endotoxin solutions between 0.0005 and 10 EU/mL and tested by the rFC kit to evaluate precision, accuracy, detection and quantification limits, linearity, and robustness. Each of the six points was assayed at least five times.The relative standard deviation for precision testing ranged from 1.9 to 8.3%. The recovery accuracy values of endotoxin were between 61% and 125% for the range from 0.005 to 10 EU/mL. The results demonstrated that the rFC method allows endotoxin quantification with accuracy, precision, specificity, and linearity for the range of 0.005 and 10 EU/mL for 0.9% sodium chloride injection. (AU)

A contaminação por endotoxinas é uma ameaça à segurança dos produtos farmacêuticos, especialmente dos medicamentos parenterais. Qualquer produto farmacêutico estéril e/ou livre de pirogênios requer especificações regulatórias para garantir a segurança de uso para o paciente. Este estudo abrange o estudo de avaliação de desempenho empregando o kit comercial Endozyme II® Go para quantificação de endotoxina, por Fator C recombinante (FCr), em amostras de cloreto de sódio 0,9% para uso parenteral. As amostras foram fortificadas com cinco concentrações distintas de soluções de endotoxina na faixa entre 0,0005 e 10 UE/mL. Cada um dos cinco níveis foi testado pelo menos cinco vezes para avaliação dos critérios de precisão, exatidão, limites de detecção e quantificação, linearidade e robustez. O desvio padrão relativo para os testes de precisão variou de 1,9 a 8,3%. Os valores de recuperação de endotoxina para o parâmetro exatidão estiveram compreendidos entre 61% e 125%. Os resultados demonstraram que o método por FCr permite a quantificação de endotoxinas com exatidão, precisão, especificidade e linearidade para a faixa de 0,005 e 10 UE/mL em amostras de cloreto de sódio 0,9% para uso parenteral. (AU)

Adsorption of aflatoxin B1 by different antimycotoxin additives: bentonite, clinoptilolite, and beta-glucans extracted from yeast cell wall.

The present study aims to evaluate and compare antimycotoxin additives (AMAs) composed of bentonite (AMA 1), clinoptilolite (AMA 2), and beta-glucans extracted from yeast cell wall (AMA 3), with respect to their ability to bind Aflatoxin B1 (AFB1) using the isothermal models of Freundlich, Langmuir, and BET. The additives were submitted to an in vitro adsorption experiment with AFB1 (0.05-4 mg L-1), using solutions of pH 3 and pH 6, with an inclusion rate of 0.5%, and analyzed by HPLC-MS/MS. At pH 3, for the seven concentrations evaluated, AMA 1 obtained adsorption rates (99.69 to 99.98%) higher (p < 0. 05) than the other AMAs, which were from 82.97 to 88.72% (AMA 2) and from 79.43 to 89.32% (AMA 3). At pH 6, in concentrations of 1, 2, and 4 mg L-1 of AFB1, AMA 1 obtained higher (p < 0.05) adsorption results (97.86 to 99.86%) than AMA 2 (91.98 to 96.12%) and AMA 3 (87.56 to 93.50%). The Freundlich model best fitted the AMA 1 adsorption data. For the other additives, the Langmuir model obtained the best fit, demonstrating qm of 8.6 mg g-1 at pH 3 and 2.3 mg g-1 at pH 6 for AMA 2; and for AMA 3, with qm of 3.4 mg g-1 at pH 3 and 2.3 mg g-1 at pH 6. The isotherm models work as an effective tool to describe the adsorption process whereas the AMA adsorption capacity varies as a function of product composition, pH, and mycotoxin content.

Determination of Novel SARS-CoV-2 Inhibitors by Combination of Machine Learning and Molecular Modeling Methods.

INTRODUCTION: Within the scope of the project, this study aimed to find novel inhibitors by combining computational methods. In order to design inhibitors, it was aimed to produce molecules similar to the RdRp inhibitor drug Favipiravir by using the deep learning method. METHODS: For this purpose, a Trained Neural Network (TNN) was used to produce 75 molecules similar to Favipiravir by using Simplified Molecular Input Line Entry System (SMILES) representations. The binding properties of molecules to Viral RNA-dependent RNA polymerase (RdRp) were studied by using molecular docking studies. To confirm the accuracy of this method, compounds were also tested against 3CL protease (3CLpro), which is another important enzyme for the progression of SARS-CoV-2. Compounds having better binding energies and RMSD values than favipiravir were searched with similarity analysis on the ChEMBL drug database in order to find similar structures with RdRp and 3CLpro inhibitory activities. RESULTS: A similarity search found new 200 potential RdRp and 3CLpro inhibitors structurally similar to produced molecules, and these compounds were again evaluated for their receptor interactions with molecular docking studies. Compounds showed better interaction with RdRp protease than 3CLpro. This result presented that artificial intelligence correctly produced structures similar to favipiravir that act more specifically as RdRp inhibitors. In addition, Lipinski's rules were applied to the molecules that showed the best interaction with RdRp, and 7 compounds were determined to be potential drug candidates. Among these compounds, a Molecular Dynamic simulation study was applied for ChEMBL ID:1193133 to better understand the existence and duration of the compound in the receptor site. CONCLUSION: The results confirmed that the ChEMBL ID:1193133 compound showed good Root Mean Square Deviation (RMSD), Root Mean Square Fluctuation (RMSF), hydrogen bonding, and remaining time in the active site; therefore, it was considered that it could be active against the virus. This compound was also tested for antiviral activity, and it was determined that it did not delay viral infection, although it was cytotoxic between 5mg/mL-1.25mg/mL concentrations. However, if other compounds could be tested, it might provide a chance to obtain activity, and compounds should also be tested against the enzymes as well as the other types of viruses.

Early Blood Clot Detection Using Forward Scattering Light Measurements Is Not Superior to Delta Pressure Measurements.

The occurrence of thrombus formation within an extracorporeal membrane oxygenator is a common complication during extracorporeal membrane oxygenation therapy and can rapidly result in a life-threatening situation due to arterial thromboembolism, causing stroke, pulmonary embolism, and limb ischemia in the patient. The standard clinical practice is to monitor the pressure at the inlet and outlet of oxygenators, indicating fulminant, obstructive clot formation indicated by an increasing pressure difference (ΔP). However, smaller blood clots at early stages are not detectable. Therefore, there is an unmet need for sensors that can detect blood clots at an early stage to minimize the associated thromboembolic risks for patients. This study aimed to evaluate if forward scattered light (FSL) measurements can be used for early blood clot detection and if it is superior to the current clinical gold standard (pressure measurements). A miniaturized in vitro test circuit, including a custom-made test chamber, was used. Heparinized human whole blood was circulated through the test circuit until clot formation occurred. Four LEDs and four photodiodes were placed along the sidewall of the test chamber in different positions for FSL measurements. The pressure monitor was connected to the inlet and the outlet to detect changes in ΔP across the test chamber. Despite several modifications in the LED positions on the test chamber, the FSL measurements could not reliably detect a blood clot within the in vitro test circuit, although the pressure measurements used as the current clinical gold standard detected fulminant clot formation in 11 independent experiments.