Abstract: This study aimed to investigate the genetic polymorphisms in Agouti signaling protein (ASIP) and melanocortin 1 receptor (MC1R) genes and their association with coat color in native Bulgarian sheep breeds. A total of 247 individuals belonging to 7 native sheep breeds from 38 herds were genotyped for the ASIP and MC1R genes. Five single nucleotide polymorphisms (SNPs) were de-tected in the MC1R gene and three haplotypes representing MC1R ED and E+ alleles were identified among the studied sheep breeds. It was found that the dominant ED allele of MC1R is absent in the breeds - Patch-faced Maritza and Dubenska. The most common haplotype (h3) of the domi-nant ED allele was associated with either black or brown coat color in native Karnobatska, Kara-kachanska and Cupper-red Shumenska breeds, respectively. We can infer that haplotype 3 alleles could be a possible result that could interpret the mechanisms of black wool colour manifestation in the native Karnobatska sheep breed, which has formed the genetic pool of this sheep breed.

Abstract:

The increasing interest in developing silver nanoparticles as antibiotic raw materials has attracted much attention, as the most common reduction and electrolysis techniques produce the toxic gas byproduct nitrogen dioxide. This paper reports a successful effort to develop a modified toxic-free electrolysis technique to produce electrolytic silver nanoparticles (ESN). A comparison of the physical and biological properties of ESN and reductive silver nanoparticles (RSN) was made. The presence of silver atoms in the solution was determined using a UV visible spectrometer and absorption peaks were found at 425 nm (ESN) and 437 nm (RSN). The particle size in solution was determined using dynamic light scattering and the diameter was found to be approximately 40 nm (for ESN) and 70 nm (for RSN). Antibacterial efficacy and power to prevent the development of bacterial resistance against Propionibacterium acnes (P. acnes) were assessed using the Kirby-Bauer method. Statistical analysis of clear zone diameter data showed that unlike RSN, the efficacy of ESN increased with higher concentrations. The efficacy of ESN and RSN is relatively lower than Chloramphenicol 5% because it is measured in different concentration units (ESN and RSN in ppm and Chloramphenicol in %). By using a calibration curve, the efficacy of 5% Chloramphenicol can be equated to 0.005% ESN. In addition, P. acnes developed strong resistance to Chloramphenicol, weak resistance to RSN and showed no resistance to ESN. These findings underscore the extraordinary potential of ESN as a raw material for future antibiotics.

Abstract: This study evaluated the effect of copaiba oil (COP) or sodium monensin, used as nutritional additives for beef cattle. Five steers cannulated in the rumen were assigned to a 5 x 5 Latin square design to the COP treatments: Control - (0 g of COP), 1.25 g COP, 2.50 COP, and 3.75 g COP per kg-1 dry matter (DM) or monensin (concentrate with the addition of 40 mg kg-1 of sodium monensin (DM basis) of total diet. The COP did not affect intake and digestibility. Monensin decreased (P ≤ 0.05) DM and nutrients intake; however, it did not affect digestibility compared to control treatment. Conversely, the monensin supply provided lower (P ≤ 0.05) mean pH values and, increases in the concentrations of N-NH3 and propionate (mmol L-1). The increased propionic acid production resulted in higher (P ≤ 0.05) proportions (mmol 100 mmol-1) of propionate, and reduction in the molar proportions of acetate and butyrate, with a consequent reduction in the acetate:propionate ratio. Copaiba oil in 1.25 to 3.75 g kg-1 does not alter the ruminal metabolism of cattle. However, sodium monensin reduces DM intake and decreases the acetate:propionate ratio in the rumen fluid.

Abstract: Growing demands for environmentally-friendly and sustainable crop pest management increased the interest for biopesticides as an alternative to the synthetic chemical pesticides. This review presents the current status of bioacaricides defined as commercial biopesticide products based on microorganisms (microbial acaricides) and biologically active substances of microbial, plant or animal origin (biochemicals and semiochemicals), used in crop protection against spider mites (Tetranychidae) and other plant-feeding mites. The most important microbial bioacaricides are mycopesticides, products manufactured from living propagules of Beauveria bassiana s.l. and several other acaropathogenic fungi. Products based on avermectins and milbemycins, secondary metabolites of actinomycetes, are well-known examples of biochemicals of microbial origin. Among biochemicals of plant origin, the most widely used have been the products based on pyrethrum, obtained from the Dalmatian daisy, Tanacetum cinerariifolium (Asteraceae), and azadirachtin, obtained from the Indian neem tree, Azadirachta indica (Meliaceae). In the recent years, products based on the essential oils from aromatic plants belonging to the families Lamiaceae, Myrtaceae, Rutaceae and others, have also gained increasing importance in the market. Special emphasis in this review is given on the compatibility of bioacaricides with predatory mites from the family Phytoseiidae as natural enemies used in the integrated management of plant-feeding mites.

Abstract: The Wnt signaling pathway is critical in the onset and progression of gastrointestinal (GI) cancers. Anomalies in this pathway, often stemming from mutations in critical components such as adenomatous polyposis coli (APC) or β-catenin, lead to uncontrolled cell proliferation and survival. In the case of colorectal cancer, dysregulation of the Wnt pathway drives tumor initiation and growth. Similarly, aberrant Wnt signaling contributes to tumor development, metastasis, and resistance to therapy in other GI cancers, such as gastric, pancreatic, and hepatocellular carcinomas. Targeting the Wnt pathway or its downstream effectors has emerged as a promising therapeutic strategy for combating these highly aggressive GI malignancies. Here, we review the dysregulation of the Wnt signaling pathway in the pathogenesis of GI cancers and further explore the therapeutic potential of targeting the various components of the Wnt pathway. Furthermore, we summarize and integrate the preclinical evidence supporting the therapeutic efficacy of potent Wnt pathway inhibitors with completed and ongoing clinical trials in GI cancers. Additionally, we discuss the challenges of Wnt pathway targeted therapies in GI cancers to overcome these concerns for effective clinical translation.

Abstract: Two related P-type ATPases designated as ATPase1 and ATPase3 have been identified in Plasmodium falciparum. These two ATPases exhibit very similar gene and protein structures and are most similar to P5B-ATPases. There are some differences in the predicted substrate-binding sites of ATPase1 and ATPase3 which suggest different functions for these two ATPases. Orthologues of ATPase3 were identified in all Plasmodium species including the related Hepatocystis and Haemoproteus. ATPase3 orthologues could also be identified in all apicomplexan species, but no clear orthologues were identified outside of the Apicomplexa. In contrast, ATPase1 orthologues were only found in the Laverania, avian Plasmodium species, and Haemoproteus. ATPase1 likely arose from a duplication of the ATPase3 gene early in the evolution of malaria parasites. These results support a model in which early malaria parasites split into two clades. One clade consists of mammalian malaria parasites and Hepatocystis but excludes P. falciparum and related Laverania. The other clade includes Haemoproteus, avian Plasmodium species, and Laverania. This contrasts to recent models which suggest all mammalian malaria parasites form a phylogenetic group and all avian malaria parasites form a separate phylogenetic group. ATPase1 may be a useful taxonomic/phylogenetic character for the phylogeny of Haemosporidia.

Abstract: Medulloblastoma (MB) is the most common malignant brain tumor in children, typically arising during infancy and childhood. Despite multimodal therapies achieving a response rate of 70% in children older than 3 years, treatment remains challenging. Ferroptosis, a form of regulated cell death, can be induced in medulloblastoma cells in vitro using erastin or RSL3. Using two independent medulloblastoma RNA-sequencing cohorts (MB-PBTA and MTAB-10767), we investigated the expression of ferroptosis-related molecules through multiple approaches, including Weighted Gene Co-Expression Network Analysis (WGCNA), molecular subtype stratification, protein-protein interaction (PPI) networks, and univariable and multivariable overall survival analyses. A prognostic expression score was computed based on a cross-validated ferroptosis signature. In training and validation cohorts, regulation of the ferroptosis transcriptional program distinguished the four molecular subtypes of medulloblastoma. WGCNA identified nine gene modules in the MB tumor transcriptome; five correlated with molecular subtypes, implicating pathways related to oxidative stress, hypoxia, and trans-synaptic signaling. One module, associated with disease recurrence, included epigenetic regulators and nucleosome organizers. Univariable survival analyses identified a 45-gene ferroptosis prognostic signature associated with nutrient sensing, cysteine and methionine metabolism, and trans-sulfuration within one-carbon metabolism. The top ten unfavorable ferroptosis genes included: CCT3, SNX5, SQOR, G3BP1, CARS1, SLC39A14, FAM98A, FXR1, TFAP2C, and ATF4. Patients with a high ferroptosis score showed worse prognosis, particularly in G3 and SHH subtypes. The PPI network highlighted IL6 and CBS as unfavorable hub genes. In a multivariable overall survival model, which included gender, age, and molecular subtype classification, the ferroptosis expression score was validated as an independent adverse prognostic marker (hazard ratio: 5.8; p-value = 1.04 × 10⁻⁹). This study demonstrates that regulation of the ferroptosis transcriptional program is linked to medulloblastoma molecular subtypes and patient prognosis. A cross-validated ferroptosis signature was identified in two independent RNA-sequencing cohorts, and the ferroptosis score was confirmed as an independent and adverse prognostic factor in medulloblastoma.

Abstract: Natural products and synthetic small molecules have been extensively studied as potential therapeutics against Mycobacterium tuberculosis (Mtb), particularly drug-resistant strains. The caseinolytic protein C1 (ClpC1) has emerged as a promising drug target for combating Mtb, and accurate measurement of its ATPase activity is essential for elucidating the mechanisms of drug candidates. In this study, we optimized a highly sensitive ATPase assay for ClpC1 and compared the enzymatic activity of various constructs, including N-terminal His6-SUMO-tagged, C-terminal His6-tagged, native full-length (FL) ClpC1, and smaller domains. Our results demonstrated that the N-terminal tag significantly impairs ClpC1 activity, whereas the C-terminal tag has no such effect. Additionally, smaller domains exhibited markedly reduced enzymatic activity compared to FL-native ClpC1. Taking the native ClpC1 forward, we tested three natural products—rufomycin (RUF), ecumicin (ECU), and cyclomarin A (CYMA)—and observed varying degrees of ATPase activity enhancement. ECU and five of its analogs were further evaluated, yielding AC50 values consistent with their binding affinities (KD) measured by surface plasmon resonance (SPR). RUF and CYMA exhibited strong KD values of 0.023 µM and 0.006 µM, respectively, and minimal inhibitory concentrations (MICs) of 0.02 µM and 0.094 µM, respectively. However, their ATPase activity enhancement was modest (<93%). In contrast, ECU and its analogs significantly enhanced ATPase activity (up to 830%) despite weaker MICs and KD values compared to RUF and CYMA. These findings suggest that RUF/CYMA and ECU may operate via distinct mechanisms of action, offering valuable insights for the development of ClpC1-targeting therapeutics against tuberculosis.

Abstract:

Degenerative proteinopathies constitute a set of molecular diseases that are caused by the misfolding of specific proteins, leading them to change their biochemical configuration and become toxic for entire systems of organs. Such protein toxicity induces the lysing of an increasing number of proteins that have a biochemically ‘wild-type’ version, gradually and eventually leading to a complete shift in the ratio between such ‘wild-type’ and ‘altered’ versions of such proteins, which directly precedes the clinical onset of such diseases. Proteinopathies not only involve neurodegenerative illnesses, but also a disease that leads to a progressive rate of blindness. Sadly, all such impairments that are neurodegenerative in nature may only receive palliative treatment, given that they are caused by aggregated proteins that start damaging and destroying entire neuronal systems, which leads to impairments in the neuro-muscular and ultimately to the inability of the patients to perform vital functions, like breathing and deglutition. There is neither a cure, nor a definitive method in which the progression of the illness can be stopped at the present time. Consequently, all neurodegenerative diseases have mortality rates of 100% and clinical approaches aim to reduce the suffering of such patients. Nonetheless, there seems to be a glimmer of hope regarding future prophylactic approaches that could delay the onset of many types of proteinopathies. Namely, an immune application could support efforts of clinical suffering delay and attenuation in an unprecedented manner. At the same time, it is necessary to emphasise upon realistic scenarios, that it remains virtually impossible to delay the onset of proteinopathies to the point of the patient reaching the average number of years in life expectancy without experiencing clinical symptoms yet. Initially, clinicians developed and tested a nasal spray containing a substance known as protollin, which stimulates a restricted extent of adaptive lymphocyte recruitment and transport to the central nervous system areas affected by initial stages of protein aggregation, activating a substantial number of microglial cells and preventing the lysis of numerous astrocytes, which in turn start lysing a number of beta-amyloid protein aggregates together without inducing pathophysiology, given the stage in which the patients have not experienced any clinical manifestation of the neurodegenerative disease yet. In case of an unsuccessful attempt to bring protollin above the threshold levels of clinical safety and efficacy, an immunostimulatory and immunomodulatory substance containing a low concentration of a mixture of recombinant Type I & III Interferons, innate and adaptive lymphocytes, perhaps themselves priorly treated with such IFN glycoproteins, would probably remain a vital candidate for an effective, yet probably still restricted delay of onset of various proteinopathies that could be neurodegenerative and optically degenerative. An existent success rate of the clinical test allows the opening of a window of opportunity regarding an increased efficacy of such adaptive lymphocyte approach, by including recombinant Type I and Type III Interferons into such a nasal spray, which could also enter adaptive lymphocyte and further improve their structural integrity and their multi-lateral functionality. Moreover, a low dose of protollin, Type I Interferons and Type III Interferons could be inserted in combination into adaptive T-Lymphocytes to optimise their defence mechanisms and immune functions, potentially bringing a considerable immunising effect against microbial diseases like HIV-induced, retroviral AIDS. Such an approach could create a stable and wide “highway bridge” of connection between innate and adaptive immunity, aiming for the best version of an immune contribution toward a considerable delay of proteinopathy clinical onset. Overall, there may be a requirement for a bi-lateral update of immunological research covering therapeutics and vaccine development; an immune system based optimisation that builds a stable and wide bridge of connection more directly between pre-cytokine and post-cytokine immune activation, and overall between innate and adaptive immune departments; and a pathogen-based optimisation that either eliminates or partially activates genes suppressive of Type I and Type III Interferon-encoding genes, helping enliven the concept of “United Immune System” as well, though less directly than the immune system-based potential approach.

Abstract:

Regulatory T cells (Tregs) play a central role in immune regulation and tolerance. The transcription factor FOXP3 is a master regulator of Tregs in both humans and mice. Mutations in FOXP3 lead to the development of IPEX syndrome in humans and the scurfy phenotype in mice, both of which are characterized by fatal systemic autoimmunity. Additionally, Treg dysfunction and FOXP3 expression instability have been implicated in non-genetic autoimmune diseases, including graft-versus-host disease, inflammatory bowel disease, rheumatoid arthritis, and multiple sclerosis. Recent investigations have explored FOXP3 expression in allergic diseases, revealing Treg alterations in food allergies, asthma, and atopic dermatitis. This review examines the multifaceted roles of FOXP3 and Tregs in health and various pathological states including autoimmune disorders, allergic diseases, and cancer. Additionally, this review focuses on the impact of recent technological advancements in facilitating Treg-mediated cell and gene therapy approaches, including CRISPR/Cas9-based gene editing. The critical function of FOXP3 in maintaining immune homeostasis and tolerance to both self-antigens and alloantigens has been emphasized. Considering the potential involvement of Tregs in allergic diseases, pharmacological interventions and cell-based immunomodulatory strategies may offer promising avenues for developing novel therapeutic approaches in this field.