Activity In Cultures Research Articles

Implementasi Gerakan Literasi Sekolah Melalui Pemanfaatan Majalah Dinding Digital

Reading and writing as one of the literacy activities in a positive school culture becomes boring. It is challenging to increase students' interest in literacy. To embody Ki Hadjar Dewantara's philosophy, learning notices the nature of the times, the school literacy movement must utilize technology advances. This research aims to increase students' interest in reading and writing and to determine digital media's effectiveness in increasing literacy ability. This is a qualitative research approach. The data collection techniques are through observation and interview. The research subjects are fourth to sixth-grade students. First, they make a wall magazine, next teacher uploads it to Google Sites, then other students watch by the link provided. The research result shows that students get more enthusiastic than looking for reference sources to write. When they place it on a digital media with varied digital displays, student’s reading enthusiasm increases. Google Sites use as a form of digital media is suitable as an alternative solution for increasing students’ interest in reading and writing. It is also effective in increasing literacy ability. The research benefit provides real experiences and a meaningful learning process. Digital media provides easy access everywhere and makes publication widely spread.

Sex-dimorphic glucose transporter-2 regulation of cAMP-protein kinase A (PKA) C-alpha pathway activity and phosphorylation in rat hypothalamic primary astrocyte cultures.

Brain astrocyte glycogenolysis is regulated in part by the second messenger adenosine 3'5'-cyclic monophosphate (cAMP). Hypothalamic astrocyte glycogen metabolism shapes glucose counterregulation, under the control of glucose transporter-2 (GLUT2), a plasma membrane glucose carrier and sensor. Hypothalamic astrocyte cAMP is subject to neurotransmitter control, but effects of nutrient cues on this messenger are unclear. Here, an established hypothalamic primary astrocyte culture model and gene knockdown tools were used to investigate the premise that GLUT2 exerts sex-dimorphic regulation of cAMP-protein kinase A (PKA) signalling in these glia. Data show that basal cAMP was elevated in female versus male; GLUT2 gene silencing up-regulated or down-regulated this profile in male versus female. Glucoprivation increased cAMP content in astrocytes of each sex, yet GLUT2 siRNA pretreatment exacerbated (male) or blunted (female) this stimulatory effect. PKA and phosphoPKA levels in glucose-supplied astrocytes were increased (male) or decreased (female) by GLUT2 knockdown. PKA protein was amplified, yet phosphoPKA was refractory to glucose withdrawal in male, while females showed sustained PKA expression alongside diminished phosphoPKA. GLUT2 siRNA pretreatment exacerbated glucoprivic augmentation of PKA content in male but down-regulated both PKA and phosphoPKA proteins in female. Evidence for parallel GLUT2 siRNA-associated changes in cAMP and PKA, albeit in opposing directions in the two sexes, infers that GLUT2 control of hypothalamic astrocyte cAMP-dependent PKA signalling is sex-specific. Data also disclose that in the female, GLUT2 curbs the baseline phosphoPKA/PKA expression ratio but is not involved in glucoprivic suppression of this ratio.

The Effect of the Optogenetic Stimulation of Astrocytes on Neural Network Activity in an In Vitro Model of Alzheimer's Disease.

Optogenetics is a combination of optical and genetic technologies used to activate or, conversely, inhibit specific cells in living tissues. The possibilities of using optogenetics approaches for the treatment of epilepsy, Parkinson's and Alzheimer's disease (AD) are being actively researched. In recent years, it has become clear that one of the most important players in the development of AD is astrocytes. Astrocytes affect amyloid clearance, participate in the development of neuroinflammation, and regulate the functioning of neural networks. We used an adeno-associated virus carrying the glial fibrillary acidic protein (GFAP) promoter driving the optogenetic channelrhodopsin-2 (ChR2) gene to transduce astrocytes in primary mouse hippocampal cultures. We recorded the bioelectrical activity of neural networks from day 14 to day 21 of cultivation using multielectrode arrays. A single optogenetic stimulation of astrocytes at 14 day of cultivation (DIV14) did not cause significant changes in neural network bioelectrical activity. Chronic optogenetic stimulation from DIV14 to DIV21 exerts a stimulatory effect on the bioelectrical activity of primary hippocampal cultures (the proportion of spikes included in network bursts significantly increased since DIV19). Moreover, chronic optogenetic stimulation over seven days partially preserved the activity and functional architecture of neuronal network in amyloidosis modeling. These results suggest that the selective optogenetic activation of astrocytes may represent a promising novel therapeutic strategy for combating AD.

Metal-Free CO Prodrugs Activated by Molecular Oxygen Protect against Doxorubicin-Induced Cardiomyopathy in Mice.

Carbon monoxide has been extensively studied for its various therapeutic activities in cell cultures and animal models. Great efforts have been made to develop noninhalational approaches for easy and controlled CO delivery. Herein, we introduce a novel metal-free CO prodrug approach that releases CO under near-physiological conditions. CO from the quinone-derived CO prodrugs is initiated by general acid/base-catalyzed tautomerization followed by oxidation by molecular oxygen to form the key norbornadienone intermediate, leading to cheletropic CO release only in an aerobic environment. Representative CO prodrug analog QCO-105 showed marked anti-inflammatory effects and HO-1 induction activity in RAW264.7 macrophages. In a mouse model of doxorubicin-induced cardiomyopathy, we show for the first time that the CO prodrug QCO-105 prevented cardiomyocyte injury, consistent with the known organ-protective effects of HO-1 and CO. Overall, such a new CO prodrug design serves as the starting point for developing CO-based therapy in attenuating the cardiotoxicity of doxorubicin.

Impact of TiO2, ZnO, and Ag nanoparticles on anammox activity in enriched river Nile sediment cultures: unveiling differential effects and environmental implications

BackgroundThe increasing use of nanoparticles (NPs) necessitates investigation of their impact on wastewater treatment processes, particularly anammox, a critical biological nitrogen removal pathway. This study explored the effects of short-term exposure to TiO2, ZnO, and Ag-NPs on anammox activity in enriched cultures derived from River Nile sediments.Materials and methodsAnammox bacteria were identified and enriched, with activity confirmed through 16S rRNA and hydrazine oxidoreductase (hzo) gene amplification and sequencing. Activity assays demonstrated efficient ammonium removal by the enriched culture. Subsequently, the impact of different sized and concentrated NPs on anammox activity was assessed.ResultsXRD analysis confirmed NP behavior within the microcosms: TiO2 transformed, ZnO partially dissolved, and Ag remained ionic. hzo gene expression served as a biomarker for anammox bacterial activity. Interestingly, 100 nm TiO2-NPs up-regulated hzo expression, potentially indicating a non-inhibitory transformed phase. Conversely, ZnO and Ag-NPs across all sizes and concentrations significantly down-regulated hzo expression, suggesting detrimental effects. Ag-NPs amended microcosms showed a significant reduction (79%) in hzo gene expression and a detrimental effect on bacterial populations. Overall, anammox activity mirrored hzo expression patterns, with TiO2 (21 and 25 nm, respectively) exhibiting the least inhibition, followed by ZnO and Ag-NPs.ConclusionThis study highlights the differential effects of NPs on anammox, with the order of impact being Ag > ZnO > TiO2. These findings provide valuable insights into the potential environmental risks of NPs on anammox-mediated nitrogen cycling in freshwater ecosystems.

Teachers’ Teaching Competence in Sport Dance Classes in a Chinese Cultural Activity Center: Basis for a New Pedagogical Framework Model

This paper explores the significance of teaching satisfaction within the realm of sports dance education, focusing on the learning environment of the Shijiazhuang Youth Science and Technology Culture Activity Center. With the rapid modernization of China's education system, there is an increasing emphasis on the integration of cultural knowledge and physical education. Sports dance, having been introduced to China in 1984, has gained popularity among teenagers for its blend of gentleman culture and competitive sports. Despite its deep mass foundation and widespread interest, the current quality of sports dance training in China is uneven, with a notable lack of professional teachers. This study aims to identify the factors influencing students' satisfaction with their sports dance teachers' instruction, and in doing so, enhance the overall learning experience and teaching effectiveness. By analyzing the teaching satisfaction survey of sports dance students, the paper reveals the main issues affecting the learning process and proposes suggestions to address these concerns, thereby contributing to the sustainable development of the sports dance community and the professional growth of its educators. The study underscores the pivotal role of teachers in shaping students' competencies and attitudes, not only in dance but also in fostering a comprehensive educational experience that aligns with the progressive expectations of modern society.

Polyphasic Characterization of the Biocontrol Potential of a Novel Strain of Trichoderma atroviride Isolated from Central Mexico.

This work describes the characterization of Trichoderma atroviride strain CMU-08, isolated from Michoacán, Mexico. CMU-08 demonstrated robust growth and conidiation across a temperature range from 16 to 32 °C and a pH range from 4 to 9 on potato dextrose agar (PDA) and malt extract agar (MEA) media. The strain is an efficient antagonist of six species of phytopathogenic fungi and oomycetes in PDA, MEA, and Vogel minimal medium (VMM). Antagonist mechanisms of CMU-08 included direct mycoparasitism observed in dual-culture assays, as well as antibiosis attributed to growth inhibition via both volatile and non-volatile metabolites, with the effectiveness varying depending on the test phytopathogen and culture medium. Extracellular filtrates (ECFs) recovered from liquid cultures of CMU-08 under basal and induced conditions using Botrytis cinerea cell walls significantly inhibited their growth at a concentration of 750 µg/mL. Moreover, in detached tomato leaf assays, these ECFs reduced foliar damage caused by B. cinerea by 24-34%. The volatile organic compounds (VOCs) produced by CMU-08 also exhibited substantial efficacy, reducing foliar damage by up to 50% in similar tests. Despite showing no basal extracellular chitinase enzymatic activity, CMU-08 demonstrated significant induction of this activity in cultures supplemented with B. cinerea and Fusarium sp. cell walls. Four genes encoding extracellular chitinases (chit33, chit36, ech42, and locus 217415) showed different dynamics of transcriptional regulation during the dual-culture confrontation of strain CMU-08 with B. cinerea and Fusarium sp., varying according to the phytopathogen and the interaction stage. The CMU-08 strain shows physiological versatility and employs a variety of antagonist mechanisms toward different species of phytopathogenic microorganisms, making it a good candidate for developing a biocontrol product for field application.

Influence of storage time of fermented soghurt on the changes in fat and protein biomolecules through Confocal Laser Scanning Microscopy (CLSM) under refrigeration condition

The aim of the study is to assess the significance of storage duration on fermented Soghurt (soy curd), focusing on changes in fat and protein biomolecules analyzing with CLSM, viscosity measurements, and particle size distributions under refrigerated conditions. Soghurt prepared with 1% yogurt culture (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) was analyzed for storage stability under refrigerated conditions for up to 15 days measuring the pH, acidity, microbial counts, viscosity, particle size distribution and scanning electron microscopy for structural analysis. Even though lactic counts were not affected significantly during the storage periods, yeast & mold counts and coliform counts were absent up to 15 days during the storage. However, the product could be stored upto 7 days under refrigeration condition (6–8 °C). During the storage period, it was observed that the viscosity of soghurt increased with increase in storage time. Particle size distribution was estimated during the entire storage periods for soghurt and it was observed that smaller particle was mostly found in the range of 3.0–50.0 µm while bigger particles were in the range of 57.0–910.0 µm. Further, larger aggregates with big particle sizes were observed during prolonged storage period. The CLSM study showed the appearance of proteins and fats being dispersed slowly during the storage due to the proteolytic and lipolytic activities of yoghurt cultures after 5 days of storage under refrigeration condition.

За полемиката по въпроси на езиковата култура в Унгария

The paper presents and comments on different opinions and views on the activity of cultivating the Hungarian standard language, also with regard to the relationship between linguistics and speech culture in the Hungarian linguistic and educational tradition. Disagreements over the place of language culture in linguistic research, the role of linguists in language cultivation, the cult of the exemplary standard language, the evaluаtion of linguistic innovations, the unity of the literary-linguistic norm across the entire Hungarian linguistic territory, etc. reveal the complexity and intricacy of the issues related to language culture activity and codification practice, as well as the influence of political, ideological and cultural-historical factors. These discussion questions shed light on current problems concerning changes in the speech practice of the contemporary Hungarian language community, the state of the Hungarian national language and literary-linguistic norms, the need to introduce new methods and techniques into the education system, etc. in order to conduct an adequate and effective language policy. The success of the country’s overall language strategy depends on the correct formulation and solution of the outlined problems

CYP5122A1 encodes an essential sterol C4-methyl oxidase in Leishmania donovani and determines the antileishmanial activity of antifungal azoles

Visceral leishmaniasis is a life-threatening parasitic disease, but current antileishmanial drugs have severe drawbacks. Antifungal azoles inhibit the activity of cytochrome P450 (CYP) 51 enzymes which are responsible for removing the C14α-methyl group of lanosterol, a key step in ergosterol biosynthesis in Leishmania. However, they exhibit varying degrees of antileishmanial activities in culture, suggesting the existence of unrecognized molecular targets. Our previous study reveals that, in Leishmania, lanosterol undergoes parallel C4- and C14-demethylation to form 4α,14α-dimethylzymosterol and T-MAS, respectively. In the current study, CYP5122A1 is identified as a sterol C4-methyl oxidase that catalyzes the sequential oxidation of lanosterol to form C4-oxidation metabolites. CYP5122A1 is essential for both L. donovani promastigotes in culture and intracellular amastigotes in infected mice. CYP5122A1 overexpression results in growth delay, increased tolerance to stress, and altered expression of lipophosphoglycan and proteophosphoglycan. CYP5122A1 also helps to determine the antileishmanial effect of antifungal azoles in vitro. Dual inhibitors of CYP51 and CYP5122A1 possess superior antileishmanial activity against L. donovani promastigotes whereas CYP51-selective inhibitors have little effect on promastigote growth. Our findings uncover the critical biochemical and biological role of CYP5122A1 in L. donovani and provide an important foundation for developing new antileishmanial drugs by targeting both CYP enzymes.

Evaluating peripheral blood lymphocyte subset composition and lipopolysaccharide-induced cytokine secretory activity in mononuclear leukocyte cultures from patients with febrile and meningeal forms of tick-borne encephalitis

Introduction.Multiple studies on immune response in patients with various clinical forms of tick-borne encephalitis (TBE) have shown conflicting results. The study aim was to estimate the changes in peripheral blood lymphocyte subset counts and activity of spontaneous and lipopolysaccharide (LPS)-stimulated cytokine production in the mononuclear leukocyte cultures in patients with febrile and meningeal acute TBE.Materials and methods.Groups 1 and 2 included 16 and 12 patients with febrile and meningeal acute TBE, respectively. The control group included 13 healthy donors. Hemogram and T-lymphocyte, T-helper cell, T-cytotoxic and NK cell counts were analyzed by flow cytometry at week 1 of the disease as well as the spontaneous and LPS-stimulated secretion levels of TNFα, IL-1β, IL-6, IL-10, IL-8, and MCP-1 were assessed by ELISA in the supernatants of mononuclear cell cultures twice: during hospitalization and two weeks later. Statistical analysis was performed by the Mann–Whitney U-test, and Wilcoxon test.Results.Group 2 demonstrated significant decrease in spontaneous and/or LPS-stimulated levels of proinflammatory cytokines IL-1β, IL-6, MCP-1, and TNFα, but showed higher IL-8 level as compared with Group 1. In addition, spontaneous and/or LPS-induced levels of IL-6 and TNFαin Group 2 did not significantly differ from the controls, which presumably also indicated the suppression of their production. In contrast, spontaneous and/or LPS-induced levels of IL-1β, IL-6, MCP-1, and TNFαin Group 1 were higher than in the controls. The spontaneous IL-10 level in the patients from both groups were higher than in the controls. Peripheral blood lymphocyte and T-cytotoxic T-lymphocyte counts in both groups of TBE patients were lower than in the controls. There was significant increase in neutrophil counts, decrease in NK cell count in Group 2 as compared to the patients with a milder febrile form.Conclusion.Meningeal acute TBE patients was presumably associated with inadequate immune response with NK cell deficiency, T-cell dysfunction, increased neutrophil count in the peripheral blood and impaired pro-inflammatory cytokine production related to innate immune response.

Green synthesis and characterization of silver nanoparticles using Oroxylum indicium leaf extract and its elicitation activity in tissue culture

Green synthesis and characterization of silver nanoparticles using Oroxylum indicium leaf extract and its elicitation activity in tissue culture

Impact of zinc oxide nanoparticles on biosynthesis of thymoquinone in cell cultures of Nigella sativa

The rising market interest for Nigella sativa (Black seeds) necessitates the development of cultivation strategies to enhance metabolites production. Zinc oxide nanoparticles (ZnO-NPs) have drawn global attention as efficient and bio safe elicitors for in vitro cultures, to enhance secondary metabolites production in medicinal plants. In this study, ZnO-NPs were utilized for establishment of callus and cell cultures in black seeds for the first time. Hypocotyl explants were cultured on Murashige and Skoog (MS) media with varying levels of ZnO-NPs, resulted in callus induction and biomass formation. Optimal response in callus growth parameters were observed when explants were grown on MS media supplemented with 60 mg/L ZnO-NPs, resulting in 71.2 % callus induction frequency, 28.2 g/L fresh biomass, 9.7 g/L dry biomass, and 63 % water content. A substantial increase in callus growth was observed when ZnO-NPs were combined with 6-Benzylaminopurine (BA) at ratio of 45:1.5 mg/L, resulting in 91.2 % callus induction frequency and 42.2 g/L fresh biomass. In cell suspension cultures, ZnO-NPs alone at 45 mg/L produced optimum callus biomass (60.9 g/L). However, in combination with BA, callus biomass did not increase significantly in cell cultures. Maximum accumulation of total phenolic content (TPC: 26.8 mg GAE/g DW; Gallic acid equivalent dry weight) and total flavonoid content (TFC: 19.5 mg QE/g DW; Quercetin equivalent dry weight) was observed in cell cultures treated with higher concentration (70 mg/L) of ZnO-NPs in the 5th week of the growth curve. Moreover, ZnO-NPs incremented substantially the Phenylalanine lyase (PAL), Superoxide dismutase (SOD) and Peroxidase (POD) enzyme activities in cell cultures. Nonetheless, Reverse Phase High Performance Liquid Chromatography (RP-HPLC) analysis indicated peak production of thymoquinone (168.5 mg/g FW) in cell cultures treated with 45 mg/L ZnO-NPs alone. This study offers a promising approach for commercial production of Nigella sativa biomass and bioactive metabolites.

Bio-based polylactic acid labware as a sustainable alternative for microbial cultivation in life science laboratories

Single-use plastics (SUPs) in life science laboratories account for approximately 5.5 million tonnes of waste per year globally. Of SUPs used in life science laboratories, Petri dishes, centrifuge tubes, and inoculation loops are some of the most common. In order to reduce the reliance on petrochemical-based SUPs in the life science research laboratory and minimize the negative environmental impacts associated with SUPs, this research investigates the applicability of polylactic acid (PLA) in single-use labware as a replacement for petrochemical-based plastics. PLA is one of the most well-studied biodegradable plastics that can be produced from sustainable resources. Commercially available PLA was used to 3D print a select range of labware to test the suitability of PLA-based material for routine microbiology work. An injection moulded PLA-based Petri dish was also designed and produced, for increased optical clarity. The biocompatibility was tested against Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus epidermidis) strains of bacteria. The PLA-based labware did not negatively impact the cell growth, viability, and metabolic activity of the bacterial cultures. The injection moulded PLA Petri dish showed a reduced colony forming unit count for the Gram-negative E. coli strain compared to the polystyrene Petri dish, ∼1.5 × 109 CFU/mL and ∼3.0 × 109 CFU/mL respectively, during late-exponential growth. The colony counts were, however, in the same order of magnitude. This observed difference may be due to the internal environment inside the Petri dish, hence the internal O2 concentration, humidity, and temperature during bacterial growth were investigated. This work demonstrates, for the first time, a full successful workflow of bacterial growth using a sustainable bioplastic, providing a pathway to reducing the environmental impacts of SUPs in life science laboratories.

Thermostable bFGF Improves Cell Lifespan by Enhancing Cell Activity in the Long-Term Culture of Human Orbicularis Oculi Stem Cells.

Stem cells derived from human orbicularis oculi muscle (hOOM) are a valuable resource for cell therapy. However, when stem cells are continuously cultured, their abilities tend to deteriorate over time. One method to address this issue is to use basic fibroblast growth factor (bFGF) to maintain the stem cell functionality. The limitation is that bFGF is unstable under mammalian cell culture conditions with a half-life of only 8 hours, which poses a significant challenge to the production and maintenance of high-quality stem cells. In this study, we used thermostable bFGF (TS-bFGF) and demonstrated that hOOM-derived stem cells cultured with TS-bFGF exhibited superior proliferation, stem cell function, reduced reactive oxygen species, and cellular senescence delay effect compared to cells cultured with wild-type bFGF. Considering the pivotal role of stem cells in broad ranges of applications such as regenerative medicine and cultured meat, we anticipate that TS-bFGF, owing to its thermostability and long-lasting properties, will contribute significantly to the acquisition of high-quality stem cells.

Influence of Storage Methods on the Vitality and Growth Rate of Macrofungi

The work contains a comparative analysis of methods for storing pure cultures of macrofungi. The study used 20 species of macrofungi from various taxonomic and ecological-trophic groups. Storage was carried out using five methods: serial subculturing, storage under a layer of distilled water and three cryopreservation protocols: a protocol using blocks of agar medium, a “perlite protocol” and a “grain protocol”. For the selected cryostorage methods, various cryoprotective compounds (glycerol, trehalose) were used. Radial growth rate was used as a criterion for the state of crops. The values of the radial growth rate obtained immediately after isolation of the pure culture were chosen as the control. It has been shown that the most favorable for preserving the physiological activity of cultures are the storage method under a layer of distilled water, “perlite” and “grain” cryopreservation protocols.

Enhanced biochemical properties of soybean root nodule asparaginase through plant molecular farming compared to bacterial enzyme for cancer treatment

Asparaginase is a therapeutic enzyme used as an anticancer agent and is typically produced through microbial fermentation using organisms such as Escherichia coli and Erwinia chrysanthemi. However, this method faces challenges, including potential enzyme contamination during production, allergic reactions to the enzyme, and stability issues requiring stringent control measures. An innovative solution is the application of plant molecular farming, utilizing Rhizobium root symbiosis for asparaginase production. The objective is to optimize nodule development for asparaginase yield, characterize the enzyme's properties, and evaluate its anticancer efficacy against microbial enzyme. In our study, we established soybean root cultures and inoculated them with Bradyrhizobium japonicum to form root nodules. We evaluated eukaryotic asparaginase production at different incubation times. We purified asparaginase from the root nodule cultures and compared its physicochemical properties and anticancer activity with microbial asparaginase. Results showed that asparaginase reached maximum activity in root nodule cultures 10 days after rhizobium inoculation in the culture media. The root nodule asparaginase exhibited a high content of alpha helices and beta sheets and a low random coil. It demonstrated higher stability and activity across different pH levels and temperatures than Escherichia coli asparaginase. Additionally, root nodule asparaginase displayed better catalytic parameters and stability over time than E. coli asparaginase. Thus, root nodule asparaginase is superior to E. coli asparaginase as an anticancer agent. This ensures the root nodule asparaginase can effectively target cancer cells, enhancing the overall therapeutic outcome. This provides a renewable, cost-effective, and environmentally friendly alternative to traditional enzyme production methods.

Influence of Storage Methods on the Vitality and Growth Rate of Macrofungi.

The work contains a comparative analysis of methods for storing pure cultures of macrofungi. The study used 20 species of macrofungi from various taxonomic and ecological-trophic groups. Storage was carried out using five methods: serial subculturing, storage under a layer of distilled water and three cryopreservation protocols: a protocol using blocks of agar medium, a "perlite protocol," and a "grain protocol." For the selected cryostorage methods, various cryoprotective compounds (glycerol, trehalose) were used. Radial growth rate was used as a criterion for the state of crops. The values of the radial growth rate obtained immediately after isolation of the pure culture were chosen as the control. It has been shown that the most favorable for preserving the physiological activity of cultures are the storage method under a layer of distilled water, "perlite" and "grain" cryopreservation protocols.

Oncolytic activity of a coxsackievirus B3 strain in patient-derived cervical squamous cell carcinoma organoids and synergistic effect with paclitaxel

BackgroundCervical squamous cell carcinoma (CSCC) is a prevalent gynecological malignancy worldwide. Current treatments for CSCC can impact fertility and cause long-term complications, underscoring the need for new therapeutic strategies. Oncolytic virotherapy has emerged as a promising option for cancer treatment. Previous research has demonstrated the oncolytic activity of the coxsackievirus B3 strain 2035 A (CVB3/2035A) against various tumor types. This study aims to evaluate the clinical viability of CVB3/2035A for CSCC treatment, focusing on its oncolytic effect in patient-derived CSCC organoids.MethodsThe oncolytic effects of CVB3/2035A were investigated using human CSCC cell lines in vitro and mouse xenograft models in vivo. Preliminary tests for tumor-selectivity were conducted on patient-derived CSCC tissue samples and compared to normal cervical tissues ex vivo. Three patient-derived CSCC organoid lines were developed and treated with CVB3/2035A alone and in combination with paclitaxel. Both cytotoxicity and virus replication were evaluated in vitro.ResultsCVB3/2035A exhibited significant cytotoxic effects in human CSCC cell lines and xenograft mouse models. The virus selectively induced oncolysis in patient-derived CSCC tissue samples while sparing normal cervical tissues ex vivo. In patient-derived CSCC organoids, which retained the immunohistological characteristics of the original tumors, CVB3/2035A also demonstrated significant cytotoxic effects and efficient replication, as evidenced by increased viral titers and presence of viral nucleic acids and proteins. Notably, the combination of CVB3/2035A and paclitaxel resulted in enhanced cytotoxicity and viral replication.ConclusionsCVB3/2035A showed oncolytic activity in CSCC cell lines, xenografts, and patient-derived tissue cultures and organoids. Furthermore, the virus exhibited synergistic anti-tumor effects with paclitaxel against CSCC. These results suggest CVB3/2035A could serve as an alternative or adjunct to current CSCC chemotherapy regimens.

Antifungal activity’s study of the of new derivatives of pyrrolo[3,4-c]pyrazol-3-ones and pyrazol-3- carboxamides in the biofilm model of Candida spp.

The search for new antifungal drugs is current interest due to the wide spread of fungal infections. To simulate the antifungal effect of new promising compounds with high antifungal activity in planktonic culture in clinical practice, it is necessary to study their effect on the biofilm of micromycetes.Aim: To study the antifungal activity of new promising representatives of silver salts of pyrazoles and their condensed systems under conditions of biofilm formation.Material and Methods. To study the antifungal activity of silver salts I and II, the micromethod of two-fold serial dilutions was used. Activity against a typical and 14 clinical highly virulent isolates of C. albicans was studied. The study of the antimycotic activity of substances under biofilm conditions was carried out using resazurin to quantify the degree of biofilm formation. The minimum inhibitory concentrations for biofilms were calculated (sMIC50 - the concentration of the antifungal substance at which a 50% decrease in fluorescence is observed compared to the positive control).Results and Discussion. It was shown that the antifungal effect of the studied compounds in biofilm culture of clinical strains of Candida spp. is significantly lower than in planktonic culture. High antifungal activity of the silver salt of pyrazole-3- carboxamide in planktonic and biofilm cultures of resistant strains of Candida albicans, exceeding the effect of the reference drug fluconazole by 2.8–11.2 times, was revealed.Conclusion. A promising pyrazole-3-carboxamide derivative has been identified that effectively inhibits the growth of Candida albicans biofilms, which can be recommended for further study.