Obesity is considered a major risk factor for the development and progression of knee osteoarthritis (OA). Apart from the mechanical effect of obesity via increase in mechanical overload of weight-bearing joints, an association with hand OA has been observed. There has been increasing interest in the role of adipokines in the pathogenesis of OA in the recent years. It has been suggested that their systemic effects link obesity and OA. In this regard, the aim of the current study was measurement and analysis of serum levels of leptin and resistin in patients with knee OA with different body mass index (BMI). Seventy-three patients with primary symptomatic knee OA at the age between 35 and 87 years (mean age 66 years) were included in the study (67 women and 6 men). The patients were from 2nd to 4th radiographic stage according to Kellgren–Lawrence scale. 43 patients were with concomitant obesity (BMI ≥ 30 kg/m², mean values 38.34 ± 8.20) and 30 patients with BMI < 30 kg/m² (mean values 25.07 ± 2.95). Eleven individuals with different BMIs, including cases with obesity but without radiographic knee OA, were examined as a control group. Serum levels of leptin and resistin were measured via ELISA method. In patients with knee OA and BMI ≥ 30 kg/m², serum levels of leptin (39.546 ± 12.918 ng/mL) were significantly higher as compared with healthy individuals (15.832 ± 16.531 ng/mL, p < 0.05) and the patients with low BMI (p < 0.05). In patients with BMI < 30 kg/m² the levels of leptin (13.010 ± 10.94 ng/mL) did not differ significantly from the respective values in the control group (p = 0.48). Serum levels of resistin were also higher in knee OA patients in comparison with healthy controls, but the difference was statistically significant only for patients with high BMI (2.452 ± 1.002 ng/mL in the group with BMI ≥ 30 kg/m²; 2.401 ± 1.441 ng/mL in patients with BMI < 30 kg/m²; 1.610 ± 1.001 ng/mL in the control group, p < 0.05). A correlation was found between the serum levels of leptin and radiographic stage of OA, i.e., higher leptin levels were present in the more advanced 3rd and 4th radiographic stage, while for resistin a correlation was observed in the patient subgroup with BMI < 30 kg/m². Serum leptin and resistin levels and clinical characteristics were analyzed in patients with different clinical forms of OA. Novel clinical correlations have been found in the current study in patients with isolated knee OA vs. cases with presence of other disease localizations. It has been observed that patients with isolated knee OA were significantly younger and had higher BMI as compared with cases in whom OA is combined with other localizations i.e., spondyloarthritis ± presence of hip OA and with generalized OA. This supports the hypothesis that presence of obesity promotes earlier development of knee OA as an isolated localization of the disease in younger patients before appearance of osteoarthritic changes at other sites. The levels of leptin and resistin in isolated knee OA were also higher. Serum levels of leptin and resistin in combination with patients’ clinical characteristics suggest existence of different clinical and laboratory profile through which more precise definition of metabolic phenotype of knee OA would be possible. Considering the fact that obesity is a modifiable risk factor that has an impact on progression of knee OA, different approaches to influence obesity may offer potential for future disease-modifying therapeutic interventions.

The general aim of this study was to investigate the negative short-term effects of different concentrations of chlorpyrifos (CPF) and cypermethrin (CYP), based on the EU legislation (MAC-EQS) in common carp (Cyprinus carpio Linnaeus, 1758) under laboratory conditions and to compare their toxicity. The fish were exposed to the pesticides for 96 h and then different histological and biochemical biomarkers were investigated in the gills and liver, and bioaccumulation analyses were conducted. The chemical studies showed increased pesticide concentrations in the gills as the first site for pollutants compared to the liver at the 96th hour. In addition, the histological analyses showed severe alterations in the gills and liver after exposure to both tested pesticides. In the gills, we found mainly intense proliferative and, to a lesser extent, degenerative changes and alterations in the circulatory system, such as necrosis and vasodilation. In the liver, regressive and progressive lesions, as well as circulatory disturbances and inflammation, were observed. The regressive lesions showed a higher degree of expression compared to the other changes. Furthermore, we found altered enzymatic activities—catalase, glutathione reductase, and glutathione peroxidase—in the liver, compared to the control. Overall, both tested pesticides impacted the studied biomarkers in common carp, even at concentrations lower than those permitted by law. However, the results of the comparative analysis showed a relatively higher toxicity of CYP compared to CPF in the fish. Still, questions persist as to whether the observed changes are adaptive or entirely destructive. To avoid any danger or risk, these pesticides must be applied cautiously, especially near water bodies.

Osteoarthritis (OA) is the most common degenerative joint disease causing progressive damages of the cartilage and subchondral bone, synovial inflammation, and severe pain. Despite the complex pathomorphological changes that occur in OA, the approach to different forms of OA is standardized. The global results from pharmacological treatment are not satisfactory. Hence, this study aimed to explore the effects of metformin, alendronate, and their combination on OA development and progression in mice with collagenase-induced osteoarthritis (CIOA). Female ICR (CD-2) mice were randomized to five groups: control group, CIOA untreated, CIOA + metformin, CIOA + alendronate, and CIOA + metformin + alendronate. OA was induced by the intra-articular (i.a.) injection of collagenase. OA phenotype was analyzed by flow cytometry (bone marrow cell differentiation), ELISA (serum levels of the adipokines leptin and resistin), and histology (pathological changes of the knee joint). Treatment with metformin, alendronate, or their combination inhibited the expression of RANK and RANKL on osteoblasts and osteoclasts obtained by ex vivo cultivation of bone marrow cells in mineralization or osteoclastogenic media. In addition, metformin treatment was effective for the attenuation of fibroblast differentiation, but not of mesenchymal stem cells (MSCs), while alendronate had an opposite effect. The combination of metformin and alendronate had a suppressive effect on both MSCs and fibroblasts differentiation. Treatment with metformin, alendronate, and their combination decreased serum concentrations of leptin and resistin in the chronic phase of arthritis. The histopathological examination showed that compared with the untreated CIOA group (OA score 9), the groups treated with metformin (OA score 4) or alendronate (OA score 6) had lower scores for cartilage changes. Metformin combined with alendronate significantly decreased the degree of cartilage degeneration (OA score 2), suggesting that this combination might be a useful approach for the treatment of OA patients.

Subchondral bone that has intense communication with the articular cartilage might be a potential target for pharmacological treatment in the early stages of osteoarthritis (OA). Considering the emerging data about the role of adipokines in the pathogenesis of OA, the administration of drugs that influence their level is also intriguing. Metformin and alendronate were administered in mice with collagenase-induced OA (CIOA) as a monotherapy and in combination. Safranin O staining was used for the assessment of changes in subchondral bone and articular cartilage. Before and after treatment, serum levels of visfatin and biomarkers of cartilage turnover (CTX-II, MMP-13, and COMP) were assessed. In the current study, the combined administration of alendronate and metformin in mice with CIOA led to the protection against cartilage and subchondral bone damage. In mice with CIOA, metformin led to a decrease in visfatin level. In addition, treatment with metformin, alendronate, or their combination lowered the level of cartilage biomarkers (CTX-II and COMP), while the level of MMP-13 was not influenced. In conclusion, personalized combination treatment in OA according to clinical phenotype, especially in the early stages of the disease, might lead to the identification of a successful disease-modifying therapeutic protocol in OA.

This study evaluated the histopathological impact of three commonly used pesticides—pirimiphos-methyl, propamocarb hydrochloride, and 2,4-dichlorophenoxyacetic acid (2,4-D)—on the kidneys of common carp (Cyprinus carpio Linnaeus, 1758) after 96-h acute exposure. The histopathological analysis demonstrated that all three tested pesticides induced structural changes. The histopathological changes were assessed using a semi-quantitative scoring system and categorised into circulatory, degenerative, proliferative, and inflammatory alterations. While circulatory alterations were absent in all treatments, clear and statistically significant degenerative, proliferative, and inflammatory responses were recorded, which escalated with increasing pesticide concentrations. Additionally, various statistical analyses were conducted to evaluate the lesions in kidney structure and function. Before the statistical analysis, normality and variance homogeneity were assessed using the Shapiro–Wilk and Levene’s tests, respectively. Due to non-normal data distribution, non-parametric methods were applied. Hence, the non-parametric statistical methods showed distinct group-level differences in the kidney damage indices. The Kruskal–Wallis test revealed significant differences across treatments (p < 0.001), and Mann–Whitney U tests identified specific pairwise differences. The degenerative and proliferative lesions were most prominent in fish exposed to 2,4-D at 100 µg/L (IK = 34), followed by pirimiphos-methyl and propamocarb hydrochloride. Inflammatory changes were mainly observed in the pirimiphos-methyl groups. The histopathological lesions were concentration-dependent, with 2,4-D causing irreversible renal damage at higher concentrations. These findings highlight the nephrotoxic risks posed by common pesticides and validate that the use of histopathological indices, combined with robust non-parametric testing, provides a reliable approach to evaluating organ-specific pesticide toxicity. These biomarkers offer sensitive early warning indicators of environmental risk, reinforcing the suitability of common carp as a model species for ecotoxicological assessment.

Honey bees are one of the most significant pollinators and contribute to the pollination of various crops. The honey bee, Apis mellifera (Linnaeus, 1758), has unique characteristics that could be successfully used to improve biomonitoring approaches in assessing environmental interactions. Three apiaries with different rates of honey bee colony losses were included in the study—Dimovtsi, Plovdiv, and Krasnovo, Bulgaria. Male individuals (immature and mature) were collected from five colonies for each of the three apiaries and studied for histopathological changes in the gonads. The results concerning the rate of honey bee colony losses in the studied apiaries from 2022 and 2023 showed honey bee losses in the tested colonies due to queen problems, which were reported for Plovdiv, as well as the death of honey bees or a reduction in their number to a few hundred bees in the colony. The chemical analysis showed the presence of different organic substances, such as Coumaphos, DEET (N, N-diethyl-M-toluamide), Fluvalinate, and Piperonyl-butoxide, in the alive and dead honey bee samples and those of food stocks (wax, pollen, and honey) within the hives. Among the sample types, those of the dead honey bees contained the greatest variety of pesticide residues, particularly in Plovdiv and Dimovtsi, reinforcing the link between pesticide exposure and honey bee mortality. The histopathological alterations were mainly associated with the thinning of the covering epithelium of the seminiferous tubules and the detachment of the basement membrane of the seminiferous tubules. The more severe histopathological lesion, necrosis, was observed in a higher degree of expression in the drones from Plovdiv, indicating a higher pollution level in this region.