A phase III study to access the safety and efficacy of prolgolimab 250 mg fixed dose administered every 3 weeks versus prolgolimab 1 mg/kg every 2 weeks in patients with metastatic melanoma (FLAT).

Background: Prolgolimab is the first Russian PD-1 inhibitor approved for the first-line treatment of unresectable or metastatic melanoma and advanced non-small cell lung cancer. It was approved in two weight-based regimens of 1 mg/kg Q2W and 3 mg/kg Q3W, but because of re-evaluation of weight-based dosing paradigm, studying of a fixed-dose regimen was considered perspective. Methods: We conducted a multicenter, single-arm, open-label efficacy, pharmacokinetics, and safety study to obtain data that would allow the approval of the new flat dosing regimen of prolgolimab in patients with previously untreated unresectable or metastatic melanoma (BCD-100-8/FLAT, NCT05783882). The primary objective was to prove the non-inferiority of prolgolimab 250 mg Q3W versus prolgolimab 1 mg/kg Q2W for the treatment of patients with unresectable or metastatic melanoma in terms of ORR according to RECIST 1.1. Patients from the MIRACULUM study (BCD-100-2/MIRACULUM, NCT03269565) comprised a historical control group. Results: One hundred fourteen patients received prolgolimab 250 mg Q3W, and 61 patients received prolgolimab (Prolgo) 1 mg/kg Q2W (historical control). Objective response was achieved by 33.3% [95% confidence interval (CI): 24.8, 42.8] of patients in the Prolgo 250 mg group compared with 32.8% (95% CI: 21.3, 46.0) of patients in the Prolgo 1 mg/kg group. Risk difference was 0.00, 95% CI (-0.12; NA), p = 0.0082. Both regimens were well tolerated, and safety profiles were comparable. The pharmacokinetic analysis (PK) showed that the regimen with the fixed dose of 250 mg Q3W was characterized by higher PK parameters. The immunogenicity study did not detect binding antibodies to prolgolimab in any of the subjects. Conclusion: The obtained results showed that the selected fixed dosing regimen of prolgolimab 250 mg Q3W is characterized by efficacy and safety parameters comparable to that observed for the 1 mg/kg Q2W regimen.

Solubilizer of bacterial origin surfactin increases the biological activity of C60 fullerene.

Currently, there exists conflicting data regarding the biological activity of unmodified fullerene C60. Various sources report its toxicity, geroprotective activity, and potential interaction with DNA. Contradictory findings regarding the toxicity of C60 may arise from the use of toxic solvents, as well as the influence of bioavailability and bioactivity on the preparation conditions of C60 suspensions. Furthermore, the microbiota of experimental animals can impact geroprotective activity results by releasing surfactants that facilitate substance penetration through the cell membrane. In this study, we selected conditions for solubilizing fullerene C60 in a solution of surfactin, a surfactant of bacterial origin, as well as in a 2% aqueous solution of TWEEN 80, employing ultrasound. Through bioluminescent analysis using lux biosensors in Escherichia coli MG1655, we observed that C60 in surfactin reduced induced genotoxic and oxidative stress. Given that surfactin enhances membrane permeability to fullerene C60, suspensions of fullerene in designated concentrations of surfactin can be regarded as a DNA protector and antioxidant, warranting further investigation as a promising component of novel drugs.

Analysis of the Current State of Research on Bio-Healing Concrete (Bioconcrete).

The relatively small tensile strength of concrete makes this material particularly vulnerable to cracking. However, the reality is that it is not always possible and practically useful to conduct studies on high-quality sealing cracks due to their inaccessibility or small opening width. Despite the fact that currently there are many technologies for creating self-healing cement composites, one of the most popular is the technology for creating a biologically active self-healing mechanism for concrete. It is based on the process of carbonate ion production by cellular respiration or urease enzymes by bacteria, which results in the precipitation of calcium carbonate in concrete. This technology is environmentally friendly and promising from a scientific and practical point of view. This research focuses on the technology of creating autonomous self-healing concrete using a biological crack-healing mechanism. The research methodology consisted of four main stages, including an analysis of the already conducted global studies, ecological and economic analysis, the prospects and advantages of further studies, as well as a discussion and the conclusions. A total of 257 works from about 10 global databases were analyzed. An overview of the physical, mechanical and operational properties of bioconcrete and their changes is presented, depending on the type of active bacteria and the method of their introduction into the concrete mixture. An analysis of the influence of the automatic addition of various types of bacteria on various properties of self-healing bioconcrete is carried out, and an assessment of the influence of the method of adding bacteria to concrete on the process of crack healing is also given. A comparative analysis of various techniques for creating self-healing bioconcrete was performed from the point of view of technical progress, scientific potential, the methods of application of this technology, and their resulting advantages, considered as the factor impacting on strength and life cycle. The main conditions for a quantitative assessment of the sustainability and the possibility of the industrial implementation of the technology of self-healing bioconcrete are identified and presented. Various techniques aimed at improving the recovery process of such materials are considered. An assessment of the influence of the strength of cement mortar after adding bacteria to it is also given. Images obtained using electron microscopy methods are analyzed in relation to the life cycle of bacteria in mineral deposits of microbiological origin. Current gaps and future research prospects are discussed.

Probiotic Bacillus amyloliquefaciens B-1895 Improved Growth of Juvenile Trout.

The aim of this study was to evaluate a new Bacillus amyloliquefaciens B-1895 probiotic as a feed additive for farmed trout. Final weight, absolute and average daily gain of fish, and average daily growth rate were higher in the group that received the probiotic than in the control group (p<0.05). Moreover, the probiotic-fed trout had more intense growth rates than the control group (higher by 15.7%; p<0.05). A decrease in feed ratio was also observed in the group that received probiotic (25% decrease; p<0.05), indicating more efficient digestion and assimilation of feed. In general, the introduction of probiotic in the feed did not adversely affect the functional status of the fish. In young trout of the control group, when assessing the general chemical composition of the organism in the muscle tissue revealed significantly (p≤0.001) higher level of moisture content by 5.1% and lower by 11.0% dry matter content. In muscle, the protein content was higher by 1.33% (p≤0.001) and fat content by 2.1% (p≤0.001) in experimental fish. Generally, Lactobacilli, Enterococcus, Vibrio, Bacillus, and coliform bacteria were found in the intestinal samples of rainbow trout. Significant reliable difference (p≤0.05) between the samples of experimental and control groups was noted in the content of Bacillus bacteria. In the control group, 5.0±0.4×103 CFU/g was detected, while in the experimental group 8.4±0.8×104 CFU/g. Overall, the data indicate that probiotic bacteria B. amyloliquefaciens B-1895 has no adverse effect on selected microorganisms in the study fish.

Fish Probiotics: Cell Surface Properties of Fish Intestinal Lactobacilli and Escherichia coli.

The properties of intestinal bacteria/probiotics, such as cell surface hydrophobicity (CSH), auto-aggregation, and biofilm formation ability, play an important role in shaping the relationship between the bacteria and the host. The current study aimed to investigate the cell surface properties of fish intestinal bacteria and probiotics. Microbial adhesion to hydrocarbons was tested according to Kos and coauthors. The aggregation abilities of the investigated strains were studied as described by Collado and coauthors. The ability of bacterial isolates to form a biofilm was determined by performing a qualitative analysis using crystal violet staining based on the attachment of bacteria to polystyrene. These studies prove that bacterial cell surface hydrophobicity (CSH) is associated with the growth medium, and the effect of the growth medium on CSH is species-specific and likely also strain-specific. Isolates of intestinal lactobacilli from fish (Salmo ischchan) differed from isolates of non-fish/shrimp origin in the relationship between auto-aggregation and biofilm formation. Average CSH levels for fish lactobacilli and E. coli might were lower compared to those of non-fish origin, which may affect the efficiency of non-fish probiotics use in fisheries due to the peculiarities of the hosts' aquatic lifestyles.

Beneficial Effects of Spore-Forming Bacillus Probiotic Bacteria Isolated From Poultry Microbiota on Broilers' Health, Growth Performance, and Immune System.

Probiotics are known for their beneficial effects on poultry health and wellbeing. One promising strategy for discovering Bacillus probiotics is selecting strains from the microbiota of healthy chickens and subsequent screening for potential biological activity. In this study, we focused on three probiotic strains isolated from the gastrointestinal tract of chickens bred in different housing types. In addition to the previously reported poultry probiotic Bacillus subtilis KATMIRA1933, three strains with antimutagenic and antioxidant properties Bacillus subtilis KB16, Bacillus subtilis KB41, and Bacillus amyloliquefaciens KB54, were investigated. Their potential effects on broiler health, growth performance, and the immune system were evaluated in vivo. Two hundred newly hatched Cobb500 broiler chickens were randomly divided into five groups (n = 40). Four groups received a standard diet supplemented with the studied bacilli for 42 days, and one group with no supplements was used as a control. Our data showed that all probiotics except Bacillus subtilis KATMIRA1933 colonized the intestines. Treatment with Bacillus subtilis KB54 showed a significant improvement in growth performance compared to other treated groups. When Bacillus subtilis KB41 and Bacillus amyloliquefaciens KB54 were applied, the most significant immune modulation was noticed through the promotion of IL-6 and IL-10. We concluded that Bacillus subtilis KB54 supplementation had the largest positive impact on broilers' health and growth performance.

Quorum-Sensing Inhibition by Gram-Positive Bacteria.

The modern paradigm assumes that interspecies communication of microorganisms occurs through precise regulatory mechanisms. In particular, antagonism between bacteria or bacteria and fungi can be achieved by direct destruction of the targeted cells through the regulated production of antimicrobial metabolites or by controlling their adaptive mechanisms, such as the formation of biofilms. The quorum-quenching phenomenon provides such a countermeasure strategy. This review discusses quorum-sensing suppression by Gram-positive microorganisms, the underlying mechanisms of this process, and its molecular intermediates. The main focus will be on Gram-positive bacteria that have practical applications, such as starter cultures for food fermentation, probiotics, and other microorganisms of biotechnological importance. The possible evolutionary role of quorum-quenching mechanisms during the development of interspecies interactions of bacteria is also considered. In addition, the review provides possible practical applications for these mechanisms, such as the control of pathogens, improving the efficiency of probiotics, and plant protection.

Antimutagenic Activity as a Criterion of Potential Probiotic Properties.

The antimutagenic activity of probiotic strains has been reported over several decades of studying the effects of probiotics. However, this activity is rarely considered an important criterion when choosing strains to produce probiotic preparations and functional food. Meanwhile, the association of antimutagenic activity with the prevention of oncological diseases, as well as with a decrease in the spread of resistant forms in the microbiota, indicates its importance for the selection of probiotics. Besides, an antimutagenic activity can be associated with probiotics' broader systemic effects, such as geroprotective activity. The main mechanisms of such effects are considered to be the binding of mutagens, the transformation of mutagens, and inhibition of the transformation of promutagens into antimutagens. Besides, we should consider the possibility of interaction of the microbiota with regulatory processes in eukaryotic cells, in particular, through the effect on mitochondria. This work aims to systematize data on the antimutagenic activity of probiotics and emphasize antimutagenic activity as a significant criterion for the selection of probiotic strains.

A Review of the Effects and Production of Spore-Forming Probiotics for Poultry.

One of the main problems in the poultry industry is the search for a viable replacement for antibiotic growth promoters. This issue requires a "one health" approach because the uncontrolled use of antibiotics in poultry can lead to the development of antimicrobial resistance, which is a concern not only in animals, but for humans as well. One of the promising ways to overcome this challenge is found in probiotics due to their wide range of features and mechanisms of action for health promotion. Moreover, spore-forming probiotics are suitable for use in the poultry industry because of their unique ability, encapsulation, granting them protection from the harshest conditions and resulting in improved availability for hosts' organisms. This review summarizes the information on gastrointestinal tract microbiota of poultry and their interaction with commensal and probiotic spore-forming bacteria. One of the most important topics of this review is the absence of uniformity in spore-forming probiotic trials in poultry. In our opinion, this problem can be solved by the creation of standards and checklists for these kinds of trials such as those used for pre-clinical and clinical trials in human medicine. Last but not least, this review covers problems and challenges related to spore-forming probiotic manufacturing.

Influence of the luxR Regulatory Gene Dosage and Expression Level on the Sensitivity of the Whole-Cell Biosensor to Acyl-Homoserine Lactone.

Aliivibrio fischeri LuxR and Aliivibrio logei LuxR1 and LuxR2 regulatory proteins are quorum sensing transcriptional (QS) activators, inducing promoters of luxICDABEG genes in the presence of an autoinducer (3-oxo-hexanoyl-l-homoserine lactone). In the Aliivibrio cells, luxR genes are regulated by HNS, CRP, LitR, etc. Here we investigated the role of the luxR expression level in LuxI/R QS system functionality and improved the whole-cell biosensor for autoinducer detection. Escherichia coli-based bacterial lux-biosensors were used, in which Photorhabdus luminescensluxCDABE genes were controlled by LuxR-dependent promoters and luxR, luxR1, or luxR2 regulatory genes. We varied either the dosage of the regulatory gene in the cells using additional plasmids, or the level of the regulatory gene expression using the lactose operon promoter. It was shown that an increase in expression level, as well as dosage of the regulatory gene in biosensor cells, leads to an increase in sensitivity (the threshold concentration of AI is reduced by one order of magnitude) and to a two to threefold reduction in response time. The best parameters were obtained for a biosensor with an increased dosage of luxRA. fischeri (sensitivity to 3-oxo-hexanoyl-l-homoserine lactone reached 30-100 pM).

Fermented Duckweed as a Potential Feed Additive with Poultry Beneficial Bacilli Probiotics.

In this study, the duckweed varieties Lemna minor, Spirodela polyrhiza, and a commercially processed duckweed food supplement were investigated as potential substrates for the propagation of two probiotic Bacillus strains, B. subtilis KATMIRA1933 and B. amyloliquefaciens B-1895. Both L. minor and S. polyrhiza were found to be suitable substrates for the propagation of both bacilli, with 8.47-9.48 Log CFU/g and 10.17-11.31 Log CFU/g after 24 and 48 h growth on the substrates, respectively. The commercial duckweed product was a less favorable substrate, with growth reaching a maximum of 7.89-8.91 CFU/g after 24 h with no further growth after 48 h. Growth and adherence of the bacilli to the three products were confirmed via electron microscopy. These strains have demonstrated health-promoting benefits for poultry and thereby have the potential to enhance duckweed as an animal feed through the process of fermentation. Duckweed has been shown to be a promising alternative resource for protein and has the opportunity to become a valuable resource in multiple industries as a potential means to increase sustainability, food security, and reduce environmental impact.

Probiotic Bacilli Inhibit Salmonella Biofilm Formation Without Killing Planktonic Cells.

Salmonellosis is a foodborne infection caused by Salmonella. Domestic poultry species are one of the main reservoirs of Salmonella, which causes the foodborne infection salmonellosis, and are responsible for many cases of animal-to-human transmission. Keeping backyard chickens is now a growing trend, increasing the frequency of direct contact with the flock and, by consequence, the incidence of Salmonella infections. Bacillus subtilis KATMIRA1933 and Bacillus amyloliquefaciens B-1895 are probiotic bacilli that produce the bacteriocins subtilosin A and subtilin, respectively. The antimicrobial activity of the two strains was determined against the reference strain Micrococcus luteus ATCC 10420. The cell-free supernatant of B. subtilis KATMIRA1933 inhibited biofilm formation by Salmonella enterica subsp. enterica serovar Hadar, Salmonella enterica subsp. enterica serovar Enteritidis phage type 4, and Salmonella enterica subsp. enterica serovar Thompson by 51.1, 48.3, and 56.9%, respectively. The cell-free supernatant of B. amyloliquefaciens B-1895 inhibited the biofilm formation of these Salmonella strains by 30.4, 28.6, and 35.5%, respectively. These findings suggest that the bacillus strains may have the potential to be used as probiotics and antibiotic alternatives for the control of Salmonella in poultry. The number of planktonic cells was unaffected by treatment with the cell-free supernatant. A co-culture of the Salmonella strains with either bacilli showed no signs of growth inhibition, suggesting that it might have been quorum sensing that is affected by the two Bacillus strains.

Mechanisms of Candida Resistance to Antimycotics and Promising Ways to Overcome It: The Role of Probiotics.

Pathogenic Candida and infections caused by those species are now considered as a serious threat to public health. The treatment of candidiasis is significantly complicated by the increasing resistance of pathogenic strains to current treatments and the stagnant development of new antimycotic drugs. Many species, such as Candida auris, have a wide range of resistance mechanisms. Among the currently used synthetic and semi-synthetic antifungal drugs, the most effective are azoles, echinocandins, polyenes, nucleotide analogs, and their combinations. However, the use of probiotic microorganisms and/or the compounds they produce is quite promising, although underestimated by modern pharmacology, to control the spread of pathogenic Candida species.

Potential Probiotics Bacillus subtilis KATMIRA1933 and Bacillus amyloliquefaciens B-1895 Co-Aggregate with Clinical Isolates of Proteus mirabilis and Prevent Biofilm Formation.

A urinary tract infection (UTI) is a multi-factorial disease including cystitis, pyelonephritis, and pyelitis. After Escherichia coli, Proteus mirabilis is the most common UTI-associated opportunistic pathogen. Antibiotic resistance of bacteria and infection recurrence can be connected to biofilm formation by P. mirabilis. In this study, human and sheep isolates of P. mirabilis were investigated for antibiotic sensitivity using an antibiotic disk test. Co-aggregation of the tested potential probiotic bacilli, Bacillus amyloliquefaciens B-1895 and Bacillus subtilis KATMIRA1933, with the isolated pathogen was also evaluated. Then, the anti-biofilm activity of naturally derived metabolites, such as subtilin and subtilosin, in the bacilli-free supernatants was assessed against biofilms of P. mirabilis isolates. The isolated pathogens were sensitive to 30 µg of amikacin and 5 µg of ciprofloxacin but resistant to other tested antibiotics. After 24 h, auto-aggregation of B. amyloliquefaciens B-1895 was at 89.5% and higher than auto-aggregation of B. subtilis KATMIRA1933 (59.5%). B. amyloliquefaciens B-1895 strongly co-aggregated with P. mirabilis isolates from human UTIs. Cell-free supernatants of B. amyloliquefaciens B-1895 and B. subtilis KATMIRA1933 showed higher antimicrobial activity against biofilms of P. mirabilis isolated from humans as compared with biofilms of sheep isolates. According to our knowledge, this is the first report evaluating the anti-biofilm activity of probiotic spore-forming bacilli against clinical and animal UTI isolates of P. mirabilis. Further studies are recommended to investigate the anti-biofilm activity and the mode of action for the antimicrobial substances produced by these bacilli, subtilosin and subtilin.

Evaluation of an Industrial Soybean Byproduct for the Potential Development of a Probiotic Animal Feed Additive with Bacillus Species.

Probiotics are gaining public attention for their application in animal husbandry due to their ability to promote growth and prevent infections. Bacillus subtilis KATMIRA1933 and Bacillus amyloliquefaciens B-1895 are two spore-forming probiotic microorganisms that have been demonstrated to provide health benefits for poultry when supplemented into their diet. These strains can be propagated on a wide range of substrates, including soybean-derived byproducts from the food processing industry. Soybean-derived byproducts are often incorporated into animal feeds, but the value of an additive could potentially be increased by the addition of probiotic microorganisms, which may decrease production costs and reduce environmental impact. In this study, a soybean byproduct and a desalted version of this byproduct were evaluated as potential substrates for the growth of two probiotic bacilli species. Chemical analysis of these byproducts showed favorable carbohydrate, fat, and amino acid profiles, which were not affected by the desalting process. The desalted byproduct was further evaluated as a substrate for the growth of B. subtilis KATMIRA1933 and B. amyloliquefaciens B-1895 under solid-state conditions, and samples from this experiment were visualized by scanning electron microscopy. The results of this study indicate that the desalted soybean byproduct is a suitable substrate for the propagation of the two Bacillus species, which grew to numbers sufficient for the formulation of a probiotic animal feed additive.

Comprehensive study of nitrofuroxanoquinolines. New perspective donors of NO molecules.

The goal of present work is the study of NO releasing mechanisms in nitrofuroxanoquinoline (NFQ) derivatives. Mechanisms of their structural non-rigidity and pathways of NO donation - spontaneous or under the action of sulfanyl radicals or photoirradiation - were considered in details, both experimentally and quantum chemically. Furoxan-containing systems of the discussed type are not capable of spontaneous or photoinduced decomposition under mild conditions, and sulfanyl (radical) induced processes are the most preferable. It was shown that appropriate modification of NFQ through [3 + 2] cycloaddition and subsequent aromatization is a powerful tool to design new prospective donors of NO molecule. Two newly obtained NFQ derivatives were proven to have unusually high NO activity in full accordance with the theoretical model. We hope that these examples will encourage community to seek for new NO active molecules among cycloadducts and modified furoxanes.

Biological Effects of C60 Fullerene Revealed with Bacterial Biosensor-Toxic or Rather Antioxidant?

Nanoparticles have been attracting growing interest for both their antioxidant and toxic effects. Their exact action on cells strongly depends on many factors, including experimental conditions, preparation, and solvents used, which have contributed to the confusion regarding their safety and possible health benefits. In order to clarify the biological effects of the most abundant fullerene C60, its impact on the Escherichia coli model has been studied. The main question was if C60 would have any antioxidant influence on the cell and, if yes, whether and to which extent it would be concentration-dependent. An oxidative stress induced by adding hydrogen peroxide was measured with an E. coli MG1655 pKatG-lux strain sensor, with its time evolution being recorded in the presence of fullerene C60 suspensions of different concentrations. Optimal conditions for the fullerene C60 solubilization in TWEEN 80 2% aqueous solution, together with resulting aggregate sizes, were determined. Results obtained for the bacterial model can be extrapolated on eukaryote mitochondria. The ability of C60 to penetrate through biological membranes, conduct protons, and interact with free radicals is likely responsible for its protective effect detected for E. coli. Thus, fullerene can be considered as a mitochondria-targeted antioxidant, worth further researching as a prospective component of novel medications.

Probiotic Intake Increases the Expression of Vitellogenin Genes in Laying Hens.

A promising approach for slowing down the rate of reproductive aging is the use of probiotic bacteria as a feed additive. In the current study was investigated the influence of the intake of a potential probiotic on the follicle content and expression of vitellogenin genes (vtg1, vtg2, vtg3) in aged hens. RNA was isolated from liver samples collected from 570-day-old laying hens and gene expression levels were measured using RT-PCR. Bacillus subtilis KATMIRA1933 supplementation had a positive effect on the number of formed follicles in hens and also triggered a significant increase in the relative expression levels of vtg1, vtg2, and vtg3. A Bacillus amyloliquefaciens B-1895 enriched diet or a combination of the two strains had a modest effect on both the number of follicles and the expression of vitellogenin genes. Additionally, the study demonstrates that vitellogenin mRNA expression levels can be considered as a biomarker in a convenient approach for analyzing the hen's egg-laying ability.

SOS Response Inhibitory Properties by Potential Probiotic Formulations of Bacillus amyloliquefaciens B-1895 and Bacillus subtilis KATMIRA1933 Obtained by Solid-State Fermentation.

The ability of fermentates of two potential probiotic strains, Bacillus amyloliquefaciens B-1895 and Bacillus subtilis KATMIRA1933, to lower the SOS response in bacteria was evaluated using Escherichia coli-based Lux biosensors (pRecA-lux) and the tested bacilli fermentates obtained through solid-state fermentation. The SOS response was stimulated by the addition of ciprofloxacine. Preparations of both Bacillus fermentates demonstrated SOS-inhibitory activity (up to 54.21%). The strain КATMIRA1933 was characterized by higher SOS-inhibitory activity. The active components of the fermentates were stable against heating, proteinase, and RNase action.

The Impact of Bacillus subtilis KATMIRA1933 Supplementation on Telomere Length and Mitochondrial DNA Damage of Laying Hens.

In the current study, we performed in vivo investigation of probiotic intake influence on nuclear and mitochondrial DNA damage of hens, using quantitative PCR techniques. The probiotic supplementation to the diet of Hisex Brown hens had no significant effect on the rate of telomere shortening. After prolonged probiotic intake (225 and 445 days), the 18-21% decrease in the mtDNA lesions was detected. Since avian mitochondrial DNA damage investigations are rare, the current study of the probiotic-enriched diet's impact on the damage of the hen mitochondrial DNA is novel and highly important. The decrease of mtDNA damage is a beneficial property, which could positively affect the reproductive aging of hens. The positive impact of probiotic supplementation on hens' performance traits such as hen-day egg production, egg weight and mass, and feed conversion ratio was observed.