Absence Of Toxicity Research Articles

Utilization of Cassava Peel as Substrate for Production of Biofertilizer and Its Effect on the Growth of Millet

The quality of the carrier is critical in influencing microbial load and shelf life of biofertilizers. Cassava peels are abundant and have little economic value, making them ideal for use as biofertilizer carrier materials. The carrier material supported the growth of the test organism, thus suggesting the presence of nutrients and absence of toxicity. During isolation and culturing of bacteria, three (3) grams of soil sample was measured and diluted with 100ml of distilled water, and mixed well to get soil suspension. Ten (10) ml of the soil suspension was poured in the first test tube and shake well, 1ml of the first test tube was transferred into the second test tube containing nine (9) ml of sterile distilled water aseptically to get dilution. One (1)ml of the suspension from the second test-tube was transferred to third test tube also containing 9ml of sterile distilled water aseptically, 1ml of the soil suspension was transferred from the third test tube to the fourth test tube,1ml of the soil suspension was transferred from the fourth test tube was transferred to the fifth test tube,1ml of the soil suspension from the fifth soil suspension was also transferred to the sixth test tube, another 1ml from the sixth soil suspension was also transferred to seventh test tube aseptically. Soil sample from test tube 4, 5, and 6 was inoculated in yeast extract media and was incubated for 24 hours. The result shows that at 2DAP, 3DAP, 4DAP, 5DAP, AND 6DAP there is statistical significant difference between the treatment and control but at 7DAP, 8DAP, 9DAP AND 10DAP there is no statistical significant difference between the treatment and control but also at 11DAP there is statistical significant difference between the treatment and the control and at 10DAG, 12DAG, 14DAG, 16DAG, 18DAG, AND 20DAG there is no statistical significant difference between the treatment and control but at 22DAG there is statistical significant difference between the treatment and control at 24DAG there is no statistical significant difference between the treatment and the control. And at 26DAG there is statistical significant difference between the treatment and the control but also at 28DAG there is no statistical significant difference between the treatment and the control using fisher’s least significant difference test. Biofertilizer is a relevant alternative for disposal of this waste and even enables the act of converting wastes to wealth.

Isatin Bis-Imidathiazole Hybrids Identified as FtsZ Inhibitors with On-Target Activity Against Staphylococcus aureus.

In the present study, a series of isatin bis-imidathiazole hybrids was designed and synthesized to develop a new class of heterocyclic compounds with improved antimicrobial activity against pathogens responsible for hospital- and community-acquired infections. A remarkable inhibitory activity against Staphylococcus aureus was demonstrated for a subset of compounds (range: 13.8-90.1 µM) in the absence of toxicity towards epithelial cells and human red blood cells. The best performing derivative was further investigated to measure its anti-biofilm potential and its effectiveness against methicillin-resistant Staphylococcus aureus strains. A structure-activity relationship study of the synthesized molecules led to the recognition of some important structural requirements for the observed antibacterial activity. Molecular docking followed by molecular dynamics (MD) simulations identified the binding site of the active compound FtsZ, a key protein in bacterial cell division, and the mechanism of action, i.e., the inhibition of its polymerization. The overall results may pave the way for a further rational development of isatin hybrids as FtsZ inhibitors, with a broader spectrum of activity against human pathogens and higher potency.

Designing of a Novel Aerolysin-based Multiepitope Vaccine against Aeromonas hydrophila Isolated from Osphronemus goramy Using Reverse Vaccinology: an in Silico Approaches

Graphical Abstract Highlight Research The study aims to develop a multi-epitope vaccine (MEV) against A. hydrophila by targeting the aerolysin toxin, a key virulence factor responsible for infections in fish and humans. Computational methods identified and optimized B-cell and T-cell epitopes, focusing on their ability to trigger immune responses without causing toxicity or allergenicity. In silico simulations demonstrated that the MEV has a strong binding affinity to immune receptors like TLR-4, MHC-I, and MHC-II, indicating its potential to induce robust cellular and humoral immunity. Structural analysis of the MEV showed a stable 3D conformation, with most residues in favorable regions, ensuring stability during immune activation. The MEV could enhance disease control in aquaculture and reduce human infection risks, offering a promising solution to address antibiotic resistance and the absence of effective vaccines. Abstract Aeromonas hydrophila, gram-negative, is a major pathogen responsible for various diseases in mammals, reptiles, amphibia, and vertebrates, including fish and humans. Targeting the specific toxin aerolysin in A. hydrophila is crucial to address antibiotic resistance and the lack of adequate and protective vaccines against this intracellular pathogen. This study aimed to identify a multi-epitope vaccination (MEV) candidate targeting A. hydrophila aerolysin toxin to combat the disease effectively. Standard biochemical characterization methods and sequencing of the 16S rRNA, rpoB, and aerA genes identified the isolate AHSA1 as A. hydrophila. Subsequently, we identified B and T cell epitopes on the aerolysin protein and separately predicted MHC-I and MHC-II epitopes. The epitopes are then evaluated for toxicity, antigenicity, allergenicity, and solubility. The vaccine design integrated multi-epitope-based constructs, utilizing specialized linkers (GPGPG) and EAAAK linkers to connect epitope peptides with adjuvants in the cholera toxin B component, thereby enhancing immunogenicity. Ramachandran plots showed that 85.25% of the residues were located in the most favorable regions, which was followed by the generously allowed zone (1.30%), the additional allowed regions (10.80%), and the forbidden regions (2.65%), thus confirming the feasibility of the modeled vaccine design. Based on docking simulations, MEV had the highest binding and interaction energies with TLR-4, TLR-9, MHC-I, and MHC-II (-1081.4, -723.2, 866.2, -9043.3 kcal/mol). Based on computational modelling, we expect the Aerolysin MEV candidate design to activate diverse immune mechanisms, stimulate robust responses against A. hydrophila, and maintain safety. The significant solubility, absence of toxicity or allergic response, and minimal side effects in animal testing all contribute to the potential clinical utility of this vaccine candidate.

Gluconate solutions for nickel electrodeposition and nickel electroless plating

Nickel electroplating has found a wide range of industrial applications as a technique for creating protective and decorative coatings of metallic and non-metallic surfaces, protecting materials against corrosion at elevated temperatures in both alkaline environments and organic acid solutions, forming a sublayer for obtaining coatings of other metals on steel, enhancing the hardness and wear resistance of surfaces, as well as for improving solderability. Such coatings can be obtained from weakly acidic aqueous and weakly alkaline complex electrolytes. In this review, we analyze the available literature on complexation of nickel(+2) with D-gluconate ion CH2OH(CHOH)4COO‾, along with that on compositions and some technological characteristics of complex D-gluconate solutions for nickel electrodeposition and electroless plating. Thus, a corrosion-resistant smooth light-colored nickel coating, tightly adhered to a copper substrate, was obtained from an electrolyte with a pH level of 8, containing 53 g/dm3 of NiSO4 •6H2O, 44 g/dm3 of D-C6H11O7Na, 25 g/dm3 of H3BO3, 53 g/dm3 of (NH4)2SO4, and 0.5–3 g/dm3 of CO(NH2)2, at a temperature of 25 °C, a cathodic current density of 2.5 A/dm2 with a current efficiency of 96.4%. An electroless Ni-P(3–18 wt%) alloy coating on copper was obtained from a solution with a pH level of 9, containing 5–30 g/dm3 of NiSO4 •6H2O, 10–60 g/dm3 of D-C6H11O7Na, 5–40 g/dm3 of NaH2PO2 •H2O, 3 g/dm3 of HOOCCH2CH2COOH, 0.5 g/dm3 of CH3(CH2)11OSO3Na, 0.002 g/dm3 of Pb(CH3COO)2•3H2O, at a temperature of 90 °C and a deposition rate of up to 0.75 μm/min. The review also discusses methods for preparing D-gluconate electrolytes for nickel plating using sodium D-gluconate, D-glucono-1,5-lactone, and nickel D-gluconate. One advantage of D-gluconate nickel plating solutions consists in the absence of toxicity and low cost of D-gluconates.

Bithiazole inhibitors of PI4KB show broad-spectrum antiviral activity against different viral families

Broad-spectrum antivirals can be extremely important for pandemic preparedness. Targeting host factors dispensable for the host but indispensable for the virus can result in high barrier to resistance and a large range of viruses targeted. PI4KB is a lipid kinase involved in the replication of several RNA viruses, but common inhibitors of this target are mainly active against members of the Picornaviridae family. Herein we describe the optimization of bithiazole PI4KB inhibitors as broad-spectrum antivirals (BSAs) active against different members of the Picornaviridae, Coronaviridae, Flaviviridae and Poxviridae families. Since some of these viruses are transmitted via respiratory route, the efficacy of one of the most promising compounds was evaluated in an airway model. The molecule showed complete viral inhibition and absence of toxicity. These results pave the road for the development of new BSAs.

Turmeric Essential Oil Constituents as Potential Drug Candidates: A Comprehensive Overview of Their Individual Bioactivities.

The therapeutic properties of turmeric essential oil have been extensively documented in both preclinical and clinical studies. Research indicates that its primary active compounds are promising candidates for addressing a wide range of pathologies, exhibiting anticancer, anti-inflammation, antioxidant, cardiovascular, hypoglycemic, dermatological, hepatoprotective, neurological, antiparasitic, antiviral, insecticidal, antifungal, and antivenom activities. While numerous compounds possess similar potential applications, the isolated active constituents of turmeric essential oil stand out due to their unique pharmacological profiles and absence of toxicity. This literature review meticulously compiles and analyzes the bioactivities of these constituents, emphasizing their molecular mechanisms of action, reported pharmacological effects, and potential therapeutic applications. The aim of this review is to provide a comprehensive synthesis of currently available clinical and preclinical findings related to individual turmeric essential oil compounds, while also identifying critical knowledge gaps. By summarizing these findings, this work encourages further research into the isolated compounds from turmeric oil as viable drug candidates, ultimately contributing to the development of innovative therapeutic strategies.

New Antibacterial and Antioxidant Chitin Derivatives: Ultrasonic Preparation and Biological Effects.

This work focuses on the first use of ultrasonic phenol-ene coupling as a polymer analogous transformation. The ultrasonic reaction was introduced into chitin chemistry, resulting in the fast and convenient preparation of new water-soluble cationic chitin derivatives. Since water-soluble derivatives of fully deacetylated chitin are poorly described in the literature, the synthesis of each new type of these derivatives is a significant event in polysaccharide chemistry. Polycations, or cationic polymers, are of particular interest as antibacterial agents. Consequently, the resulting polymers were tested for their antibacterial activity and toxicity. We found that the highly substituted polymer of medium molecular weight exhibited the most pronounced in vitro antibacterial effect. We prepared nanoparticles using the ionic gelation technique. The most effective in vitro antibacterial chitin-based systems were tested in vivo in rats. These tests demonstrated outstanding antibacterial effects combined with an absence of toxicity. Additionally, we found that the resulting polymers, unlike their nanoparticle counterparts, also exhibited strong antioxidant effects. In summary, we demonstrated the effectiveness of ultrasound in polymer chemistry and highlighted the importance of the sonochemical approach in the chemical modification of polysaccharides. This approach enables the synthesis of derivatives with improved physicochemical and biological properties.

Predictive and pharmacodynamic biomarkers for combination therapy in stage III-IV melanoma: A Simon phase II trial (NCT03999749).

222 Background: The evaluation of biomarkers that predict response and immune-related adverse events (irAEs) is crucial for optimizing melanoma treatment with Immune checkpoint inhibitors (ICIs). We report predictive and pharmacodynamic biomarkers for response and irAEs using high-dimensional flow cytometry in a phase II trial combining nivolumab (3 mg/kg), ipilimumab (1 mg/ml), and the IL-6 receptor inhibitor tocilizumab (4 mg/kg) in unresectable/advanced melanoma patients. Methods: We analyzed serum and blood from melanoma patients at baseline, weeks 7 and 37, categorized them by irAE severity and clinical response. Patients were categorized based on the severity of irAEs (grades 3-5) (TOX), the absence of toxicity or grades 1-2 irAE (NT), clinical benefit (CB), and non-clinical benefit (NCB). Immune monitoring regimen included ELISA for IL-6, gp130, IL-23, osteopontin, and Luminex oncological panel (LXSAHM-22). We employed multi-parametric flow cytometry panels to examine myeloid cells, cytotoxic T cells, and checkpoint protein expression on immune cells. We processed ~1 million PBMCs per sample using various antibodies, with data acquisition performed via Sony’s ID7000 cytometer. The data underwent quality control, normalization PBMCs per patient, concatenation, dimensionality reduction, and clustering in FlowJo. Results: Among the 70 patients administered treatment, we observed a best overall response rate of 57%. Grade 3-5 irAEs occurred in 22% of patients by week 24. Patients manifesting irAEs exhibited notably higher baseline Th17 cell counts (NT=181±18 vs. Tox=311±49, p<0.01) and CD26+ Naive CD4 T-cell populations (NT=5838±801 vs. Tox=9632±1343, p<0.04), suggesting their potential role as predictive markers for toxicity. Additionally, in vitro investigations led us to identify CD26 co-stimulation as a preliminary step in TH17 cell differentiation. Key pharmacodynamic markers included a significant elevation in Treg cells (NCB=1286±71 vs. CB=1492±95, p=0.02) and reduction in serum gp130 levels (NCB=104±5 vs. CB=83±5 ng/ml, p<0.01) at week 7 in addition to IL-6 (NCB=7.5±1.5 vs. CB=4.6±1.1 pg/ml, p<0.01) and osteopontin (NCB=95±15 vs. CB=25±3 ng/ml, p<0.01) by week 37 when compared to baseline. Conclusions: Our findings suggest that Th17 cells can serve as a toxicity biomarker for combination therapy with ipilimumab, nivolumab and tocilizumab. Circulating Treg cells and serum osteopontin have emerged as promising pharmacodynamic biomarkers. Clinical trial information: NCT03999749 .

Antioxidant properties and bioactivity of three marine microalgae on human cancer cell lines

ABSTRACT Marine microalgae offer significant potential as sustainably cultivable microorganisms for the biorefining of a diverse array of functional metabolites. In this study, three species of marine microalgae, Tetraselmis chuii Butcher CCAP 66/21B (Chlorophyte), Phaeodactylum tricornutum Bohlin CCAP 1052/1 (Diatom) and Amphidinium carterae Hulburt CCAP 1102/8 (Dinoflagellate), were batch cultivated and their biomass retrieved for analysis in terms of pigment and fatty acid composition as well as bioactivity potential. Sequential solvent maceration of the biomass returned extracts was used to determine antioxidant capacity and assess their growth inhibition potential against A549 (a human lung carcinoma) and MCF-7 (a breast carcinoma) cell lines. The extracts of A. carterae showed higher antioxidant capacity (≤ 33.5 μmol Trolox Equivalent mg Recovered Dried Material‒1) compared to the other extracts. No major cytotoxic activity nor production of reactive oxygen species was observed using the extracts of T. chuii and P. tricornutum. However, the methanolic extraction of A. carterae inhibited the growth of both A549 and MCF-7 cells (< 4%) at 25, 50 and 100 µg ml−1 concentrations, as determined using the MTS assay and AO/PI cell staining. Microalgal metabolites have the potential for practical applications in a variety of industrial sectors. The absence of toxicity and substantial antioxidant potential of extracts of T. chuii and P. tricornutum confirm their suitability for incorporation into animal feeds or human nutrition supplements. A. carterae also appears to be a promising source of bioactives that can be further valorized via multi-compound biorefinery pipelines.

Формирование свойств биодеградируемых пленок на основе рыбного желатина

Nowadays in the food and nutrition industry special attention is paid to the creation of non-toxic, easily recy-clable packaging materials of a new assortment, capable of providing reliable protection of products from harmful factors. The object of the study was biodegradable film compositions based on fish gelatin and agar, with the addition of plasticizing agent - glycerol. Experimental and analytical studies were carried out using standard and original methods. Modeling of the process of thermal treatment of film compositions was carried out, the experimental dependence of moisture absorption of films on temperature (range 120-160 °C) and duration of the thermal modification process (10-30 minutes) was established. The possibility of thermal modification of biodegradable film compositions was determined. The rational mode of thermal treatment of films is the temperature interval of 140 °C for 20 minutes, which allows to keep the moisture absorption index at the maximum level (842%), exceeding the initial index by 8.4 times, during 120 hours. The established parameters of thermal structuring allow to keep organoleptic quality indicators of films at a high level and to obtain a homogeneous, porous and elastic film. Thermally modified films at temperatures of 140 and 160 °C are characterized by increased dissolution time up to 40 and 70 minutes respectively at dissolution temperature of 60 °C and up to 15 and 30 minutes respectively at dissolution temperature of 75 °C. It is shown that thermal modification of biodegradable films will contribute to compliance with safety requirements, environmental friendliness, absence of toxicity. Thermal modification allows to increase functional and technological properties of composite biodegradable films, which can be used to create a wide range of modern packaging barrier materials.

Visualization of 2D and 3D Tissue Models via Size-Selected Aqueous AgInS/ZnS Quantum Dots.

Three-dimensional (3D) spheroid cell cultures of fibroblast (L929) and tumor mammary mouse (4T1) were chosen as in vitro tissue models for tissue imaging of ternary AgInS/ZnS fraction quantum dots (QDs). We showed that the tissue-mimetic morphology of cell spheroids through well-developed cell-cell and cell-matrix interactions and distinct diffusion/transport characteristics makes it possible to predict the effect of ternary AgInS/ZnS fraction QDs on the vital activity of cells while simultaneously comparing with classical two-dimensional (2D) cell cultures. The AgInS/ZnS fractions, emitting in a wide spectral range from 635 to 535 nm with a mean size from ∼3.1 ± 0.8 to ∼1.8 ± 0.4 nm and a long photoluminescence lifetime, were separated from the initial QD ensemble by using antisolvent-induced precipitation. For ternary AgInS/ZnS fraction QDs, the absence of toxicity at different QD concentrations was demonstrated on 2D and 3D cell structures. QDs show a robust correlation between numerous factors: their sizes in biological fluids over time, penetration capabilities into 2D and 3D cell structures, and selectivity with respect to penetration into cancerous and healthy cell spheroids. A reproducible protocol for the preparation of QDs along with their unique biological properties allows us to consider ternary AgInS/ZnS fraction QDs as attractive fluorescent contrast agents for tissue imaging.

Effects of antiseptic irrigation solutions on osseointegration in a cementless tibial implantation mouse model.

Despite the success of standard antiseptic irrigation solutions in reducing periprosthetic joint infection (PJI) rates, there is still a need for more effective solutions. Synergistic use of povidone-iodine (PI) and hydrogen peroxide (H2O2) has shown promising results; however, the optimal solution concentration balancing bactericidal activity and osseointegration remains unknown. This study aims to evaluate the impact of these antiseptic irrigation solutions on osseointegration and the bone-implant interface strength in vivo. Forty C57BL/6 mice underwent bilateral tibial implantation surgery and were randomly allocated into three groups receiving 0.3% PI, 10% PI mixed with 3% H2O2, or saline as irrigation solutions intraoperatively. Assessments were performed on postoperative Days 1 and 28, including plain radiographs, microcomputed tomography (microCT) evaluation, histological analysis, immunohistochemistry, and biomechanical pull-out testing. No wound complications were observed. MicroCT scans revealed no differences in peri-implant trabecular bone parameters. Biomechanical pull-out testing showed no differences in the bone-implant interface strength across groups. Histological analysis indicated no differences in bone and bone marrow percentage areas among treatment groups. Immunohistochemical analysis demonstrated no differences among groups in peri-implant osteocalcin, osterix, or endomucin-positive cells. In conclusion, using either antiseptic irrigation solution showed no differences in osseointegration parameters compared to the control group, demonstrating safety and the absence of toxicity. CLINICAL RELEVANCE: Dilute 0.3% povidone-iodine and a 1:1 combination of 10% povidone-iodine mixed with 3% hydrogen peroxide can be safely used during primary and revision total joint arthroplasty without compromising osseointegration or causing wound complications.

Ozonation process at neutral pH integrated with anaerobic treatment system to methylparaben removal

The incomplete removal of emerging pollutants, such as parabens, from wastewater effluents represents a major pathway for their release into the environment. The development of advanced technologies integrated into well-known conventional systems is a solution. Advanced oxidative processes (AOP), such as ozonation and its combinations, are technologies based on the generation of hydroxyl radicals (HO•) capable of removing these pollutants, representing an alternative solution to this problem. Therefore, the present study aimed to investigate the ozonation process combined with hydrogen peroxide (O3/H2O2) in the treatment of anaerobic effluent post-treated by a real-scale upflow anaerobic sludge blanket (UASB) reactor, enriched with methylparaben (MeP). The study was conducted under optimized conditions and at neutral pH of the effluent. To investigate the influence of variables, as ozone concentration, hydrogen peroxide concentration, pH and time, a 24 Factorial experimental design followed by a 22 Doehlert optimization design was implemented. The optimal degradation occurred with 25.0 mg L−1 of O3 and 30.0 mg L−1 of H2O2 at neutral effluent pH, achieving a removal efficiency of 71.6 % after 60 min and 85.5 % after 120 min. The optimized O3/H2O2 process after 60 min achieved a 95.0 % removal of organic matter by biochemical oxygen demand (BOD), reducing the initial concentration from 138 mg L−1 to 7 mg L−1. Acute toxicity test with Artemia salina and Lactuca sativa, indicated an absence of toxicity (TU < 0.4). The findings of this study indicate the positive efficiency of combining post-UASB effluent treatment with O3/H2O2 processes for the removal of methylparaben.

Mechanistic insights on the antibacterial action of the kyotorphin peptide derivatives revealed by in vitro studies and Galleria mellonella proteomic analysis

ObjectivesThe selected kyotorphin derivatives were tested to improve their antimicrobial and antibiofilm activity. The antimicrobial screening of the KTP derivatives were ascertained in the representative strains of bacteria, including Streptococcus pneumoniae, Streptococcus pyogenes, Escherichia coli and Pseudomonas aeruginosa. MethodsKyotorphin derivatives, KTP-NH2, KTP–NH2–DL, IbKTP, IbKTP-NH2, MetKTP-DL, MetKTP-LD, were designed and synthesized to improve lipophilicity and resistance to enzymatic degradation. Peptides were synthesized by standard solution or solid-phase peptide synthesis and purified using RP-HPLC, which resulted in >95 % purity, and were fully characterized by mass spectrometry and 1H NMR. The minimum inhibitory concentrations (MIC) determined for bacterial strains were between 20 and 419 μM. The direct effect of IbKTP-NH2 on bacterial cells was imaged using scanning electron microscopy. The absence of toxicity, high survival after infection and an increase in the hemocytes count was evaluated by injections of derivatives in Galleria mellonella larvae. Proteomics analyses of G. mellonella hemolymph were performed to investigate the underlying mechanism of antibacterial activity of IbKTP-NH2 at MIC. ResultsIbKTP-NH2 induces morphological changes in bacterial cell, many differentially expressed proteins involved in DNA replication, synthesis of cell wall, and virulence were up-regulated after the treatment of G. mellonella with IbKTP-NH2. ConclusionWe suggest that this derivative, in addition to its physical activity on the bacterial membranes, can elicit a cellular and humoral immune response, therefore, it could be considered for biomedical applications.

TGF-β blockade drives a transitional effector phenotype in T cells reversing SIV latency and decreasing SIV reservoirs in vivo

HIV-1 persistence during ART is due to the establishment of long-lived viral reservoirs in resting immune cells. Using an NHP model of barcoded SIVmac239 intravenous infection and therapeutic dosing of anti-TGFBR1 inhibitor galunisertib (LY2157299), we confirm the latency reversal properties of in vivo TGF-β blockade, decrease viral reservoirs and stimulate immune responses. Treatment of eight female, SIV-infected macaques on ART with four 2-weeks cycles of galunisertib leads to viral reactivation as indicated by plasma viral load and immunoPET/CT with a 64Cu-DOTA-F(ab’)2-p7D3-probe. Post-galunisertib, lymph nodes, gut and PBMC exhibit lower cell-associated (CA-)SIV DNA and lower intact pro-virus (PBMC). Galunisertib does not lead to systemic increase in inflammatory cytokines. High-dimensional cytometry, bulk, and single-cell (sc)RNAseq reveal a galunisertib-driven shift toward an effector phenotype in T and NK cells characterized by a progressive downregulation in TCF1. In summary, we demonstrate that galunisertib, a clinical stage TGF-β inhibitor, reverses SIV latency and decreases SIV reservoirs by driving T cells toward an effector phenotype, enhancing immune responses in vivo in absence of toxicity.

Efficacy and safety of guttiferone E in melanoma-bearing mice.

Melanoma, an aggressive and potentially fatal skin cancer, is constrained by immunosuppression, resistance, and high toxicity in its treatment. Consequently, there is an urgent need for innovative antineoplastic agents. Therefore, this study investigated the antimelanoma potential of guttiferone E (GE). In an allogeneic murine B16 melanoma model, GE was administered subcutaneously and intraperitoneally. Antitumor evaluation included tumor volume/weight measurements and histopathological and immunohistochemical analysis. Furthermore, the toxicity of the treatments was evaluated through body/organ weights, biochemical parameters, and genotoxicity. Subcutaneous administration of 20 mg/kg of GE resulted in a significant reduction in both tumor volume and weight, effectively suppressing melanoma cell proliferation as evidenced by a decrease in mitotic figures. The tumor growth inhibition rate was equivalent to 54%. This treatment upregulated cleaved caspase-3, indicating apoptosis induction. On the other hand, intraperitoneal administration of GE showed no antimelanoma effect. Remarkably, GE treatments exhibited no toxicity, evidenced by non-significant differences in body weight gain, as well as organ weight, biochemical parameters of nephrotoxicity and hepatotoxicity, and genotoxic damage. This study revealed, for the first time, the efficacy of subcutaneous administration of GE in reducing melanoma, in the absence of toxicity. Furthermore, it was observed that the apoptotic signaling pathway is involved in the antimelanoma property of GE. These findings offer valuable insights for further exploring GE's therapeutic applications in melanoma treatment.

Optimizing Sex Ratios of Hyalella azteca to Reduce Variability in Reproduction and Improve Reproductive Toxicity Test Methods.

Standard toxicological test methods with Hyalella azteca incorporate both lethal and sublethal (growth, reproduction) endpoints, although lethal endpoints are often favored in routine testing. However, sublethal endpoints are important to consider because they are ecologically relevant and are often more sensitive than lethality. It is difficult to achieve robust data for reproduction in H. azteca because high biological variability is associated with reproductive yield, likely due to complex interactions in mate selection, which can add to the variability in brood sizes that females produce. In addition, effects on reproduction often co-occur with effects on growth, making it difficult to differentiate effects on growth from those on reproduction in standard tests initiated with juveniles. The present study characterized the reproductive capacity of H. azteca by investigating the role of sex ratios in reproductive yield. Experiments were initiated in the absence of toxicants with sexually mature (6-8-week-old) individuals that were placed in different female-to-male ratios (1:1, 2:3, 3:2, and 7:3). Reproduction was monitored during weekly static renewals for 7 weeks. The results indicated that a higher female:male ratio (seven females to three males) improved reproductive success and lowered biological variability in reproduction. In addition, the body lengths and weights of newly mature amphipods were used to create a size distribution to aid in the identification of small female amphipods. Based on the results of our study, a novel reproductive toxicity test method is proposed that begins with sexually mature amphipods in a ratio of seven females to three males to minimize reproductive variability in amphipods and to enable a more effective assessment of contaminants using reproduction as a highly sensitive endpoint. However, evaluation of this novel method with toxicants is needed to demonstrate that results are comparable to standard methods.Environ Toxicol Chem 2024;43:712-722. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Evaluation of antioxidant, toxicological and anxiolytic-like effect of ethanolic extracts of Ziziphus cotinifolia Reissek in adult zebrafish (Danio rerio)

BackgroundThe GABAergic and serotoninergic neurotransmission pathways are involved in the control of anxiety and ensure emotional balance. Some plant species have substances with anxiolytic activity, a therapeutic effect associated with the presence of phenolic compounds. MethodsBefore assessing the existence of anxiolytic activity, ethanolic extracts from leaves, branches and roots of Ziziphus cotinifolia Reissek were evaluated regarding biological activity and absence of toxicity. Later the influences of the samples on locomotor and anxiolytic activity, anxiolytic pathways and probable mechanisms of neurotransmission were evaluated. ResultsThe EEtFJ extract (0.5 mg/mL) did not have its anxiolytic activity reversed by the GABAergic receptor antagonist (Fmz + EEtFJ = EEjFJ; p<0.05). The twig extracts and flumazenil only partially reduced the time spent in the light zone (Fmz + EEtGJ < + EEtGJ; p<0.05), the root extract was not reversed by flumazenil (Fmz + EEtFRJ = EEtFRJ). ConclusionThe initial analysis data reveal the presence of phenolic compounds that explain the biological activities found and, as they act on different targets of neurotransmission systems involved in the control of anxiety disorders, they have potential for the production of herbal medicines with synergistic action.

INFLUENCE OF MODIFIED PEPTIDES FROM THE FETAL THYMUS ON THE ACTIVITY OF T-LYMPHOCYTES AND NATURAL KILLERS IN EXPERIMENTAL VIRAL HEPATITIS

The results showed that immunomodulin has a moderate regulatory effect on the spontaneous blast transformation of lymphocytes in the in vitro system. The costimulatory effect of immunomodulin in the reaction of PHA-induced blast transformation of T-lymphocytes was revealed. The data we obtained on the rather pronounced interferon-inducing properties of Sanogen developed by us and its combinations with well-known inductors -Cycloferon and Betaleukin, can be of practical use in the treatment of infectious pathology, especially viral hepatitis, when the combination of hepatoprotective, anti-inflammatory, antiviral, immunomodulatory and detoxification mechanisms of action, in the absence of toxicity and side effects, will ensure the development of sanogenetic processes in the patient's body.

Glyoxal acid-free (GAF) histological fixative is a suitable alternative to formalin: results from an open-label comparative non-inferiority study

Formalin, an aqueous solution of formaldehyde, has been the gold standard for fixation of histological samples for over a century. Despite its considerable advantages, growing evidence points to objective toxicity, particularly highlighting its carcinogenicity and mutagenic effects. In 2016, the European Union proposed a ban, but a temporary permission was granted in consideration of its fundamental role in the medical-diagnostic field. In the present study, we tested an innovative fixative, glyoxal acid-free (GAF) (a glyoxal solution deprived of acids), which allows optimal tissue fixation at structural and molecular level combined with the absence of toxicity and carcinogenic activity. An open-label, non-inferiority, multicentric trial was performed comparing fixation of histological specimens with GAF fixative vs standard phosphate-buffered formalin (PBF), evaluating the morphological preservation and the diagnostic value with four binary score questions answered by both the central pathology reviewer and local center reviewers. The mean of total score in the GAF vs PBF fixative groups was 3.7 ± 0.5 vs 3.9 ± 0.3 for the central reviewer and 3.8 ± 0.5 vs 4.0 ± 0.1 for the local pathologist reviewers, respectively. In terms of median value, similar results were observed between the two fixative groups, with a median value of 4.0. Data collected indicate the non-inferiority of GAF as compared to PBF for all organs tested. The present clinical performance study, performed following the international standard for performance evaluation of in vitro diagnostic medical devices, highlights the capability of GAF to ensure both structural preservation and diagnostic value of the preparations.