Colorectal cancer ranks third in global incidence and second in cancer mortality. Patient-derived models are irreplaceable for studying tumor biology. We established a human epithelial cell line from a rectal adenocarcinoma overexpressing cancer stem cell marker ALDH1A1, and we investigated the effect of ALDH1A1 knockout on tumor cell traits. The cell line and its CRISPR-Cas9 ALDH1A1 knockouts were characterized by genomic and cytogenetic methods (CNV, WES, RNAseq, karyotype), in vitro (proliferation, response to chemotherapy, migration, invasion, apoptosis), and in vivo methods. We identified the landscape of somatic mutations and copy number alterations in the original tumor and the derived cell line. Genetic attenuation of ALDH1A1 was characterized by an increase in migratory potential and extensive metastatic ability, accompanied by reduced growth of subcutaneous xenografts and alterations in gene expression associated with inhibited proliferation and promoted invasion and metastasis, ultimately resulting in dysregulation of the Wnt signaling pathway. Increased metastatic potential was also confirmed in HT-29 cells after ALDH1A1 genetic attenuation. CRISPR-Cas9-mediated editing led to functional, cellular, and molecular changes confirming the role of ALDH1A1 in colorectal cancer carcinogenesis.

Brown and beige adipocytes promote metabolic health through adaptive thermogenesis, yet the molecular mechanisms that govern their function remain incompletely understood. Here, we identify the Large Tumor Suppressor kinases (LATS), central regulators of the Hippo pathway, as key modulators of brown adipocyte activity and systemic energy homeostasis. We demonstrate that LATS kinases, particularly LATS2, modulate thermogenic gene expression, UCP1 protein levels and mitochondrial respiration by controlling the stability and nuclear localization of the transcriptional co-regulator TAZ. Mechanistically, silencing of LATS kinases leads to TAZ stabilization, which represses the activity of the master regulator PPARγ, impairing thermogenic activation. In vivo, brown adipocyte-specific deletion of Lats1 and Lats2 in mice reduces UCP1 protein levels and thermogenic potential during cold exposure without altering brown adipocyte identity. Remarkably, Lats1 and Lats2 deletion in brown fat suppresses food intake and protects against diet-induced obesity independently of ambient temperature or UCP1 function. Transcriptomic analysis of brown adipose tissue identifies Vnn1, Mfge8, S100b, and Angptl3, as potential BAT-derived regulators of appetite. These findings uncover the LATS–TAZ–PPARγ axis as a critical regulator of brown adipocyte thermogenesis and identify a previously unrecognized role of LATS kinases in controlling systemic energy homeostasis via non-thermogenic BAT functions.

Oats are widely included in gluten-free diets, yet the biological effects of oat prolamins (avenins) on intestinal homeostasis remain incompletely understood. Here, we compared purified wheat gliadins and oat avenins administered at equivalent concentrations in a BALB/c murine model. After five weeks of dietary exposure, no overt intestinal pathology was observed, as villous architecture and inflammatory cytokine expression remained largely unchanged. Nevertheless, both gliadin and avenin induced antigen-specific humoral responses, with avenin eliciting broader and more frequent IgG reactivity. Proteomic analysis of the small intestine revealed coordinated alterations in pathways related to epithelial homeostasis, antigen handling, and immune regulation. Reduced expression of caspase-8 and interleukin-18 suggested disrupted epithelial regulatory balance, while increased markers of antigen transport indicated enhanced mucosal antigen sampling. In parallel, antibody-responsive animals exhibited reduced IL-10 expression, consistent with impaired regulatory control. Together, these findings support a model in which dietary prolamins actively modulate mucosal regulatory networks and induce a primed mucosal state characterised by immune activation without overt inflammation. Given the substantially lower natural abundance of avenins compared with gliadins, these effects likely reflect dose-dependent responses rather than intrinsic pathogenicity. This model could additionally serve as a useful experimental platform for monitoring the combined effects of dietary additives and prolamins on intestinal barrier integrity and mucosal immune responses.

Given the global rise in antibiotic resistance, this study focuses on developing an alternative treatment for deep skin and soft tissue infections (SSTIs) caused by a carbapenem-resistant Pseudomonas aeruginosa. An injectable hydrogel composed of polyvinyl alcohol and chitosan was developed as a stable carrier for the porcine cathelicidins PR-39 and Protegrin-1. In vitro tests demonstrated that the addition of 8 mM EDTA fully restores the activity of antibacterial peptides in fetal bovine serum and eliminates the influence of divalent cations. In a porcine model, the application of this hydrogel resulted in a significant reduction in bacterial load after only two doses, with levels dropping below 105 CFU/g of tissue. In addition to eliminating the infection, the treated group showed early granulation tissue formation, reduced necrosis and inflammation, and the development of a continuous epidermis, all of which were completely absent in the untreated group. The research confirms that the combination of cathelicidins and EDTA in a hydrogel carrier represents a highly effective approach for treating complicated bacterial infections.

To address the challenge of drug accumulation and penetration at the tumor site(s), herein we describe a first-in-class nanocarrier containing 24 copies each of two bioactive peptides (BAPs) genetically fused in frame to the 24 N-termini of a human ferritin H-type construct, named THE-10. The two BAPs are specific for PD-L1 and integrin αVβ3/αVβ5 plus Neuropilin (iRGD) respectively, conferring immune checkpoint blockade and drug-internalization properties. In turn, the THE-10 backbone brings 48 BAPs contiguous for synergism, prolonged blood half-life, and release into the tumor microenvironment upon conditional cleavage of a metalloprotease-sensitive site. Predicted THE-10 multitasking activity was experimentally supported as follows. Size-exclusion chromatography and surface plasmon resonance demonstrated BAP cleavage/release and receptor binding (nanomolar KD). Live-cell/time-lapse imaging demonstrated 4-fold-increased internalization of naked therapeutic antibodies, mirrored by enhanced cytotoxicity of the corresponding Antibody-Drug Conjugate. Slight antitumor effects were observed in vivo by treating immune checkpoint-sensitive syngeneic mouse colorectal model with THE-10 alone. Drug boosting was instead considerable on colorectal and pancreatic tumor allografts when THE-10 was co-administered with both small and large chemotherapeutic agents, outperforming the original iRGD cyclic peptide. Thus, THE-10 may enhance target therapy, chemotherapy and immunotherapy altogether, e.g. it candidates as a multitasking, all-round, antineoplastic therapy booster.

In this article, we describe the course and diagnosis of pulmonary alveolar proteinosis (PAP) based on two cases from our practice. The first case is a 52-yearold woman, the second a 34-year-old man. Both referred patients were examined by a pulmonologist for interstitial lung disease, in the first case also with transition to pulmonary fibrosis. As part of the differential diagnosis, these patients were hospitalized at the NÚTPCHaHCH in Vyšné Hágy. Chest X-ray showed diffuse bilateral lung infiltrates, in the first patient locally confluent. Chest CT showed parenchymal involvement of the lungs with bilateral ground-glass opacities with thickened interlobular septa (crazy paving). Bronchoscopic examination was performed in both patients with bronchoalveolar lavage, which had a characteristic milky-glazed appearance. Videothoracoscopic lung biopsy was additionally indicated and histopathologically there were pulmonary alveolar proteinosis confirmed. Therapeutically, the patients underwent large volume lung lavage, with clinical condition improvement, including radiological findings improvement. We point out the basic pillars of the diagnosis of pulmonary alveolar proteinosis, which are the pattern of pulmonary involvement in the radiographic and CT (or HRCT) images, the characteristic appearance of the bronchoalveolar lavage fluid, and additionally also the histopathologic pattern of pulmonary involvement in this disease. We emphasize the need for centralized management of patients with lung diseases, which is particularly urgent in cases of rare diseases, where it provides rapid availability of all relevant diagnostic and therapeutic options, including large-volume lung lavage.

The progression of cancer strongly depends on the tumor microenvironment and immune surveillance. Tumor necrosis factor alpha (TNFα), a key inflammatory cytokine, can drive both tumor elimination and promotion, depending on its dose and the type of cancer. Colorectal cancer cell lines HCT 116, HT-29, and melanoma cells A375 engineered to stably overexpress the human TNFα gene were used to induce experimental subcutaneous tumors in two immunodeficient mouse strains: athymic Balb/c-nu/nu and SCID/bg mice. In athymic mice, TNFα-overexpressing cells completely lost their tumorigenicity. In SCID/bg mice, with no mature T and B cells and defective NK cells, the TNFα-overexpressing cells formed rudimentary flat ulcerous xenografts with rapidly reduced size, with tumor penetrance of 50-85%. Histopathological analysis revealed necrotic lesions, a more differentiated phenotype of tumor cells forming pseudoglandular structures, and more abundant stromal cells. Intratumoral infiltration of immune cells increased in TNFα-secreting tumors. Positivity of cytokeratins 7 and 20 in colorectal cancer xenografts was decreased. Paradoxically, the expression of ALDH1A1 and ALDH1A3 isoforms, which are important for disease prognosis, was increased. Our study suggests that careful modulation of the tumor microenvironment to a tumor-suppressive one using cytokine TNFα and controlled stimulation of antitumor immunity can contribute to the improvement of cancer treatment.

Five cases of patients with systemic connective tissue diseases (CTD) who developed connective tissue disease-associated interstitial lung disease (CTD-ILD) with progressive pulmonary fibrosis (PPF) are reported here. Unspecified ILD was diagnosed using high-resolution computed tomography (HRCT). Histologically, all cases were usual interstitial pneumonia (UIP) with findings of advanced (3/5) to diffuse (2/5) fibrosis, with a partially (4/5) to completely (1/5) formed image of a honeycomb lung. The fibrosis itself spread subpleurally and periseptally to more central parts (2/5) of the lung, around the alveolar ducts (2/5), or even without predisposition (1/5). Simultaneously, there was architectural reconstruction based on the mutual fusion of fibrosis without compression of the surrounding lung parenchyma (1/5), or with its compression (4/5). The whole process was accompanied by multifocal (1/5), dispersed (2/5), or organized inflammation in aggregates and lymphoid follicles (2/5). As a result of continuous fibroproduction and maturation of the connective tissue, the alveolar septa thickened, delimiting groups of alveoli that merged into air bullae. Few indistinctly visible (2/5), few clearly visible (1/5), multiple indistinctly visible (1/5), and multiple clearly visible (1/5) fibroblastic foci were present. Among the concomitant changes, areas of emphysema, bronchioloectasia, and bronchiectasis, as well as bronchial and vessel wall hypertrophy, and mucostasis in the alveoli and edema were observed. The differences in the histological appearance of usual interstitial pneumonia associated with systemic connective tissue diseases (CTD-UIP) versus the pattern associated with idiopathic pulmonary fibrosis (IPF-UIP) are discussed here. The main differences lie in spreading lung fibrosis, architectural lung remodeling, fibroblastic foci, and inflammatory infiltrates.

BACKGROUND:The pancreatic cancer-associated fibroblasts (pCAFs) are among the most active components of the pancreatic ductal adenocarcinoma (PDAC). The pCAFs being of mesenchymal stem/stromal cell origin, interact directly with tumor stromal elements, modulate tumor development, and are involved in the formation of pre-metastatic niches that result in unsatisfactory PDAC treatment outcomes. This study aimed to develop an innovative approach for the treatment of desmoplastic pancreatic carcinoma via intracellularly targeted exosomes derived from pCAFs and from mesenchymal stem cells (MSCs) transduced with the suicide gene - yeast cytosine deaminase::uracil phosphoribosyl transferase (yCD::UPRT). METHODS:pCAFs were isolated from four PDAC tumor specimens and MSCs from various tissues. Their transduction with yCD::UPRT gene produce homogenous gene transduced cell populations capable of secreting suicide gene exosomes. Both gene- transduced and naive cells and their exosomes underwent characterization by biophysical, biochemical, microscopic, and LC-MS/MS proteomic methods. Tumor cell-killing functionality was assessed using three pancreatic cancer cell lines. The killing efficacy of combined suicide gene exosomes of MSCs and pCAFs was measured in a mixture of pCAFs and MIA PaCa-2 cells as a simulated desmoplastic pancreatic tumor in vitro. RESULTS:MSCs and pCAFs suicide gene exosomes act as cancer cell-targeted drugs, effectively killing pancreatic carcinoma cells. Exosomes intracellular convert the non-toxic prodrug 5-fluorocytosine into cytotoxic 5-fluorouracil and its metabolites in a dose-dependent manner. In experiments simulating the desmoplastic microenvironment of PDAC, we have found that the suicide gene exosomes from both cells conjugated with prodrug effectively target and inhibit the growth of simulated PDAC. CONCLUSION:Exosomes containing the yCD::UPRT gene from pCAFs and MSCs function as "Trojan horse" therapies, efficiently and dose-dependently eliminating pancreatic cancer cells. PDAC environment-targeted yCD::UPRT-gene exosomes from MSCs and pCAFs show promise for a novel PDAC treatment.

The study aimed to identify altered signaling pathways and potential non-invasive biomarkers for pancreatic neuroendocrine tumors (PanNETs) through transcriptomic profiling of tumor tissues. The analysis encompassed samples from non-functional PanNETs (NF-PanNETs), insulinomas, and tumor-adjacent pancreatic tissues (TAPT). In the differential expression analysis comparing PanNETs and TAPTs, we identified 1,210 differentially expressed genes at a false discovery rate significance threshold of < 0.05 and with Log2FoldChange values of > 0.5 and <-0.5. Further pathway enrichment analysis revealed a multitude of overrepresented signaling pathways related to cell proliferation, survival, and tumorigenesis. Significant findings included the Beta-catenin-independent and TCF-dependent WNT signaling pathways, MAPK1/MAPK3 signaling, and terms associated with PI3K/AKT/mTOR signaling. Among the list of DEGs, we also identified 28 upregulated genes encoding cell surface proteins and 24 upregulated genes encoding cancer-associated secretome proteins. Since the proteins of these genes are found in the bloodstream, there is potential for further testing of these markers as biomarkers for liquid biopsy assays. Overall, these findings underscore the promise of transcriptomic landscape analysis in identifying PanNET-specific non-invasive biomarkers and uncovering potential therapeutic targets.

Here we report the accidental discovery of a gunshot wound involving a diabolo pellet in the liver of a cat. Toxicology results investigating the possibility of poisoning by anticoagulant rodenticide-type agents (brodifacoum, bromadiolone, difenacoum, diphethialone, flocoumafen, and warfarin) and for cholinesterase inhibitor pesticides (carbofuran) were negative. Internal examination discovered minor amounts of free blood in the chest cavity and uncoagulated blood in the heart cavities. The cardiac musculature of the ventricles is hypertrophied, and cavities of the ventricles are narrowed. The lungs exhibit an edema, and there are extensive confluent diffuse hemorrhages. The kidneys are light brown in color, soft in consistency, with dotted or diffuse hemorrhages in the cortex and marrow. There is dark red fluid in the bladder. There is a slightly protruding structure in the center of the right lateral lobe of the liver in view of the diaphragmatic surface, in which the diabolo pellet, with a wound, has been encased. Autopsy documents reveal renal failure. In relation to the autopsy findings, attention is paid to the impact of the release of lead from the diabolo pellet into the body, impairing filtration, leading to renal failure and death. In the puzzle of the cause of death based on autopsy and toxicological findings, the work is a good example of the importance of considering several factors in renal failure even without further investigation.

Chemotherapy with cisplatin (CDDP) and etoposide (ETO) is the standard treatment for gastroenteropancreatic neuroendocrine carcinoma (GEP-NEC). Despite high initial response rates, treatment-related toxicity and acquired resistance remain significant clinical challenges. No molecularly targeted therapies are currently established for this aggressive cancer. Here, we evaluated Aurora kinase inhibition with alisertib (ALI) as a synthetic lethality strategy in ARID1A-deficient GEP-NEC. Subcutaneous and orthotopic xenograft models were established in immunodeficient mice using the GEP-NEC-derived NT-38 cell line, and the therapeutic efficacy of ALI was compared with CDDP + ETO. Tumor responses were assessed by immunohistochemistry, Western blotting, and RNA sequencing. To confirm ARID1A dependency, transient knockdown was induced in the pancreatic neuroendocrine NT-18LM cell line, demonstrating that ALI sensitivity is significantly enhanced in ARID1A-deficient cells. ALI achieved antitumor efficacy comparable to chemotherapy and was well tolerated, with minimal weight loss relative to CDDP + ETO. Distant metastases, an early feature of GEP-NEC, developed in five animals over six weeks, two of which were treated with ALI and three controls. Transcriptomic profiling revealed convergence of both treatments on signal transduction, focal adhesion, receptor tyrosine kinase, and VEGFA-VEGFR2 signaling, while ALI uniquely enriched pathways related to pancreatic secretion, lipid metabolism, protein processing in the endoplasmic reticulum, and extracellular matrix organization. These findings establish Aurora kinase inhibition as mechanistically distinct and selectively effective in ARID1A-deficient GEP-NEC. Given its efficacy, favorable tolerability, and ARID1A-dependent specificity, ALI may represent a promising alternative to platinum-based chemotherapy, offering a strong rationale for further development of mechanism-driven combination strategies for GEP-NEC.

BackgroundThe dual allergen exposure hypothesis states that sensitization to food antigens occurs through a damaged skin barrier in individuals with no previous oral tolerance to certain foods. However, the resulting allergic reaction could depend on factors such as the host’s genetic predisposition as well as the skin and gut microbiota.MethodsSpecific-pathogen-free BALB/c and C57BL/6 and germ-free (GF) BALB/c mice were epicutaneously sensitized with ovalbumin (OVA) via dorsal tape-stripped skin and challenged with OVA by intragastric gavage. The development of food allergy (FA) symptoms, the Th2 and mast cell immune response and differences in the skin and gut microbiota were investigated.ResultsBALB/c mice, but not C57BL/6 mice, showed severe clinical signs of FA (hypothermia, diarrhea) as well as a stronger serum antibody response and Th2 cytokine secretion in the spleen and jejunum after OVA-treatment. The increased mast cell count correlated with higher MCPT-1 production and histidine decarboxylase mRNA expression in the jejunum of these mice. The 16S rRNA sequencing analysis revealed lower abundance of short-chain fatty acids producing bacteria in the gut microbiome of OVA-treated BALB/c mice. Changes in the β-diversity of the gut microbiome reflect both the genetic background as well as the OVA treatment of experimental mice. Compared to SPF mice, GF mice developed more severe anaphylactic hypothermia but no diarrhea, although they had a higher mast cell count, increased MCPT-1 production in the jejunum and serum, and increased arachidonate 5-lipoxygenase mRNA expression.ConclusionsWe show that the BALB/c mice are a mouse strain of choice for model of adjuvant-free epicutaneous sensitization through the disrupted skin barrier and following food allergy development. Our results highlight the significant influence of genetic background and microbiota on food allergy susceptibility, emphasizing the complex interplay between these factors in the allergic response.

The aim of this study is to examine the influence of food restriction on rabbit ovarian functions. A total of eight females were fed ad libitum (NF), while eight females were subjected to 50% food restriction (RF). One month later, all females were euthanized. Weights and lengths of ovaries and uterine horns were measured. Representative parts of the ovaries were subjected to histomorphometry analysis of folliculogenesis. Granulosa cells were isolated and cell viability, proliferation (accumulation of PCNA, cyclin B1, and BrdU-positive cells), apoptosis (accumulation of bax, caspase 3, and DNA fragmentation) were evaluated. Granulosa cells were subjected to proteomic analysis by using the nano HPLC-Chip-MS/MS method. Estradiol and progesterone release by ovarian and granulosa cells was assessed by ELISA. Ovarian and uterine horn weights were lower in RF than NF. The diameter of follicles and oocytes and the thickness of the theca and granulosa cells were higher in RF than NF. RF showed a lower percentage of cells containing bax and caspase 3, occurrence of DNA fragmented cells, and estradiol and progesterone. RF had higher incorporation of BrdU, a higher proportion of cells containing PCNA and cyclin B1, and a lower percentage of viable cells. RF produced more specific proteins than NF, including peptides involved in cell differentiation, proliferation/division, mitotic cell cycle, and GTP-ase activity. In conclusion, food restriction can activate reproduction by (1) selection of the growing primordial follicles, (2) better transformation of secondary to preovulatory follicles, (3) increasing growth of oocytes, (4) increasing proliferation and decreasing apoptosis in granulosa cells, (5) changes in ovarian secretory activity, and (6) changes in the number of peptides.

Heat stress exerts a negative impact on the production indicators of animals and, can, threaten their lives. The aim of this study is to document and describe macroscopic, as well as microscopic findings in laying hens who died because of heat stress during the summer heat. This is a sample of 14 laying hens which were selected from mass mortality events in poultry farming businesses. The cause of death was cardiorespiratory failure caused by overheating. Characteristic macroscopic findings were scalp cyanosis, skeletal muscle anaemia with a cooked-meat-like appearance, petechiae and hyperaemia of internal organs. Microscopic examination revealed venostasis, with dilated blood vessels overfilled with erythrocytes. The diagnostic findings document hyperaemia, cyanosis, a cooked meat-like appearance, noticeable venostasis of internal organs and intravascular haemolysis. These findings resulted from pathological conditions that developed gradually in a warm environment and led to death. The pathophysiology and changes that develop as a consequence of high temperatures are discussed. This article could be useful for veterinary pathologists diagnosing deaths caused by prolonged exposure to extraneous environmental high temperatures.

The present study aimed to identify the novel mechanisms regulating rabbit fecundity. For this purpose, the association between litter size variability, fecundity, ovarian morphology, and markers of proliferation, apoptosis, steroidogenesis, and the presence of regulatory proteins were examined in ovarian cells from two rabbit lines divergently selected for low (LL) and high (HL) variability in litter size throughout sixteen generations. Ovaries and uteri were isolated from multiparous non-lactating female rabbits from each line at sixteen generations of selection. One part of the ovary was subjected to histomorphometric analysis of folliculogenesis. From the rest of ovary, ovarian granulosa cells (OGCs) were isolated, cultured, and cell viability, proliferation (accumulation of PCNA and of cyclin B1, and BrdU-positive cells), and apoptosis (accumulation of caspase 3, bax and DNA fragmentation) were evaluated by the Trypan blue exclusion test and BrdU, quantitative immunocytochemistry, and cell death detection assays. Furthermore, OGCs were subjected to proteomic analysis by using the nano HPLC-Chip-MS/MS method. The release of progesterone and oestradiol was measured by ELISA. The LL had more than one kit per litter than the HL (7.6 kits vs. 6.5 kits, p <= 0.05). No differences were found in the diameter of primordial and primary ovarian follicles, theca, and granulosa thickness, but the diameter of oocytes in the primary and secondary follicles was higher in the LL than in the HL (88.49 mu m in the LL vs. 77.86 mu m in the HL for oocytes of primary follicles, p <= 0.05; 122.10 mu m in the LL vs. 109.87 mu m in the HL for oocytes of secondary follicles, p <= 0.05). Preovulatory follicles were presented only in the ovaries of the LL. The LL had higher incorporation of BrdU and reduced accumulation of bax within OGCs (0.92% in the LL vs. 0.44% in the HL for incorporation of BrdU, p <= 0.05; 41% in the LL vs. 48% in the HL for accumulation of bax, p <= 0.05). Ovarian fragments from the LL produced less progesterone and oestradiol than those of the HL (12 ng/mg tissue/day in the LL vs, 45 ng/mg tissue/day in the HL for progesterone, p <= 0.05; 11 ng/mg tissue/day in the LL vs. 30 ng/mg tissue/day in the HL for oestradiol, p <= 0.05). Besides, the OGCs from the LL produced a higher number of specific regulatory proteins involved in cell differentiation, proliferation, and adhesion than the HL (50 vs 38, p <= 0.05). In conclusion, higher prolificacy in the LL line would be caused by: (1) the selection of growing primordial ovarian follicles; (2) better transformation to preovulatory follicles; (3) increased cytoplasmic maturation of oocytes; (4) increased DNA synthesis and decreased cytoplasmic apoptosis in OGCs; (5) changes in ovarian steroidogenesis; and (6) changes in the number of peptides involved in cell differentiation, proliferation, and adhesion.HIGHLIGHTSAssessing fecundity in two divergently rabbit lines for variability in litter size.Fecundity and its variability are related to the diameter and quality of oocytes.Selection and better growth of preovulatory follicles have been related to fecundity.

Peripheral nerve sheath tumours (PNSTs) are rare in the mammary glands of dogs. Here, we report a case of a schwannoma, composed of two parts, of the mammary gland of a dog. The first part consists of clusters of uncircumscribed, alternating, more concentrated and looser regions. In the more concentrated parts, typical fascicularly arranged schwannoma intermingle in plexiform arrangement, more subtle in appearance than in neuronal tumour areas. The second part undergoes regression consisting predominantly of residual shorter rosettes of cells with the presence of a peculiar variably sized swirling of target-like formations consisting of compact, thicker, pinkish lamellae also with occasional adjacent cells. Immunohistochemically, the tumour cells are positive for evidence of vimentin and neuro-specific enolase. They exhibit the variable expression of the S-100 protein, show mild CD56 positivity, and focally mildly accentuated proliferative activity as assessed by Ki-67. The tumour elements are negative for evidence of cytokeratin 7, cytokeratin 20, and oestrogen receptors. Hybrid tumours may change their morphology in combination with atypical localisation and may be underdiagnosed in veterinary biopsy practice. They differ from epithelial tumours prognostically, as well as in their development and behaviour, therefore it is essential to clearly differentiate them.

The coronavirus pandemic has resulted in over 775 million cases and 7 million deaths worldwide, driving efforts to develop therapeutic strategies to control the viral infection. Therapeutic oligonucleotides have shown promise in treating many pathological conditions, including those of viral origin. The present study assessed the in vivo efficacy and safety of ASC1R, a novel therapeutic oligonucleotide of unconventional design targeting the conserved viral RdRp sequence essential for replication. In functional studies, ASC1R was administered to transfected C57BL/6 mice at doses of 1 and 10 mg/kg. Safety assessments included acute toxicity evaluations at doses ranging from 30 to 100 mg/kg, and subacute toxicity evaluations of repeated doses of 1 and 10 mg/kg. Evaluations included general clinical observations, findings at necropsy, measurements of organ weight, and histopathological examinations of the liver, lungs, spleen, and kidneys. ASC1R effectively reduced RdRp levels >94 % within 24 hours following a single 1 mg/kg dose, with no observed organ toxicity. Acute and subacute toxicity assessments found that mice receiving high (≥30 mg/kg) or repeated (10 mg/kg for 7 days) doses of ASC1R showed an increase in relative spleen weight, without histopathological changes. The marked ability of a single low dose of ASC1R (1 mg/kg) to reduce viral RNA suggests its potential for clinical applications, balancing therapeutic efficacy with minimal side effects. Our findings indicate that ASC1R has promise as a viable treatment option for patients with COVID-19.

INTRODUCTION:Staphylococcus aureus is a gram-positive, facultatively anaerobic coccus capable of causing infectious diseases in animals and humans. Especially dangerous are multidrug-resistant forms with poor or even no response to available treatments. OBJECTIVES:The study aimed to verify the effect of enzybiotics on the healing of S. aureus-infected skin wounds in an experimental pig model. METHODOLOGY:Two pigs were included in the experiment and wounds (10/pig) of 5 × 5 cm in size with 2 cm spacing were made by incision on their backs. The wounds were infected with a methicillin (oxacillin) and amoxicillin-resistant strain of S. aureus (MRSA). The experimental groups consisted of individual wounds that were infected with one sequence type of S. aureus at a concentration of 2 × 109 CFU/mL. Two wounds were left untreated (N), four wounds were using hydrogel with added lysostaphin, and four wounds were treated using hydrogel with added lysostaphin and endolysin. Subsequently, samples were taken from each wound on days 4, 7, 11, and 14. The material was processed using a standard histological technique of paraffin blocks and the sections were stained with hematoxylineosin. RESULTS:The results show that these defects present a full spectrum of reparative changes with re-epithelialization with alternating sections of necrosis and newly formed granulation tissue with an accompanying round cell inflammatory infiltrate in edematous tissue and surface scabs. On the surface of the wounds and also in smaller groups in the newly formed granulation tissue, coccoid formations corresponding to S. aureus are visible. Compared to untreated wounds, hydrogel dressings with added lysostaphin or lysostaphin and endolysin trapped greater numbers of S. aureus cocci colonies, which subsequently died off to a large extent. CONCLUSION:Enzybiotics may have interesting potential in the topical therapy of MRSA-infected skin wounds.