Biotechnic & Histochemistry最新文献

Corneal injuries are common in human and veterinary ophthalmology. There are many studies which have investigated the treatment of corneal epithelial defects. This study aimed to investigate the effects of Neutral Protamine Hagedorn (NPH) insulin as an ointment for wound healing in experimental corneal defects. First, a superficial keratectomy was performed on 12 rabbits using a corneal trephine. The animals were divided into two groups, Group I (treated, n = 6) and Group II (vehicle control, n = 6). Insulin ointment was applied topically once daily in the treated group, and saline ointment was applied in the same manner in the vehicle control group. Corneal defects were observed and photographed, and changes in wound surface were recorded on days 7, 14, 21, 30, and 60. In both groups, a significant reduction in the wound surface area was noticeable on the 30th day after defect creation. Between the 30th and 60th days, while the changes in the wound surface area in Group II remained limited, the decrease continued rapidly in Group I. At the end of the study, the corneal opacity score observed was lower in Group I than in Group II. In conclusion, we determined that topical NPH insulin may accelerate corneal wound healing after superficial lamellar keratectomy. A new alternative treatment may be developed for treating corneal wounds through continuing studies on this subject.

The main symptoms of depression, a chronic mental illness, include sadness, low self-esteem, and a diminished sense of enjoyment in life. Many factors have been suggested to be associated with depression, one of which is low testosterone in men. The serotonin reuptake inhibitor fluoxetine (FLU), used to treat depression, has been reported to potentially have detrimental effects on spermatogenesis in rats after long-term use. The multimodal antidepressant vortioxetine (VTX) offers new promise for the treatment of depression. The chronic unpredictable mild stress (CUMS) model is widely known as an experimental paradigm used to study depression-like behaviors in rodents. Stress leads to various neurochemical and immune changes, affecting multiple organs. Our study aims to examine the histopathological findings in testicular tissue induced by CUMS and the immunohistochemical expression of Cas-8, IL-6, and RANKL using a depression model in rats. Rats were split into 4 groups of 7 animals each at random. Group 1 (control) did not experience any stress. Group 2 (CUMS) was exposed to chronic, unpredictable mild stress using a specific procedure. Group 3 (CUMS+VTX) and Group 4 (CUMS+FLU) underwent CUMS and received intraperitoneal drug treatment at a dose of 10 mg/kg during the final three weeks of the study. The rat testicles collected during necropsy were evaluated histopathologically and immunohistochemically for Cas-8, IL-6, and RANKL expressions using a light microscope. In Group 1, histological analysis showed normal tissue architecture in the testicles and epididymis. In Group 2, there was significant depletion of spermatozoa in the seminiferous tubules and empty tubules in the epididymis. In Groups 3 and 4, FLU and VTX treatment led to improvements in the testicles. Cas-8, RANKL, and IL-6 immunohistochemistry revealed increased expression in Group 2, primarily in interstitial cells. In Groups 1, 3, and 4, no or very slight expression of these markers was observed. The results of this study showed that sperm production in the testes is negatively affected in CUMS-induced depression and that Cas-8, IL-6, and RANKL expression is increased, particularly in interstitial cells. VTX and FLU, used in the treatment of depression, suggest potential for mitigating the adverse effects of CUMS on the testes.

Dexmedetomidine has been shown to exert protective and curative effects on various tissues and organs in different pathological processes. This study aimed to investigate the effect of dexmedetomidine on the regeneration process after making holes in the parietal bones of rabbits. Twenty-four male Oryctolagus cuniculus rabbits were allocated to three groups, and an 8-mm circular parietal critical-sized bone defect was induced in each animal. Group_C (control) received saline; Group_LD (low dose) was given dexmedetomidine 2.75 µg/kg; Group_HD (high dose), dexmedetomidine 5.5 µg/kg; all were administered intraperitoneally for 7 days. After 8 weeks the bones were examined by micro-computed tomography (micro-CT) and histomorphometry. The results indicated that regeneration was improved in both the dexmedetomidine-treated groups. The lower dose increased the bone volume ratio (BV/TV) more than the higher dose. Trabecular thickness, connectivity value, and connectivity density were also higher in Group_LD than in Group_HD. Significant intramembranous ossification was observed in the dexmedetomidine-treated groups, and active osteoblasts were seen at the margins of new bone trabeculae. We conclude that dexmedetomidine, especially at the lower dosage, increases osteoblastic activity and regeneration quality.

Numerous studies reported about potential effects of L-carnosine in regulation of tumor growth and metabolism. We evaluated the effects of different concentrations of L-carnosine from Karnozin EXTRA® supplement on mitochondrial respiratory chain complexes of human embryo lung fibroblasts (MRC-5) and human breast cancer cells (MCF-7), with different energy pathways. Also, we analyzed the proliferation index and expression of various markers of oxidative stress. Treatment with Karnozin EXTRA^® (concentration of L-carnosine were 2, 5 and 10 mM) for 24 hours gradually decreased the number of cells and changed their morphological features. In both cell lines, a dose-dependent reduction of cell viability was recorded compared to the control group. Also, experimental groups showed a concentration-dependent decrease in fluorescence intensity of SOD2 expressions in MCF-7, while in MRC-5 we noticed higher fluorescence intensity in Carnosine 2 mM group. Treated cells, in both cell lines, showed different intensity of iNOS cytoplasmic immunopositivity in a concentration-dependent manner. In all experimental groups, we noticed an increased expression of marker of oxidative stress-cytochrome P450 2E1 (CYP2E1). The effects of Karnozin EXTRA® capsule on mitochondrial respiration, assessed with the Clark-type electrode, were manifested as a reduction of: basal cell respiration, maximum capacity of electron transport chain and mitochondrial ATP-linked respiration. Also, significant decrease in the activity of complex I (NADH-ubiquinone oxidoreductase), complex II (succinate dehydrogenase) and complex IV (cytochrome c oxidase) was observed in both cell lines. Bearing in mind that Karnozin EXTRA® is a potential regulator of energy metabolism of MCF-7 and MRC-5, these results provide a good basis for further preclinical and clinical research.

Anterior cruciate ligament (ACL) injury and subsequent reconstruction induce marrow adipose tissue (MAT) accumulation accompanied by bone loss. Short-term immobilization or non-weightbearing after ACL reconstruction further promotes MAT accumulation. However, it is unclear if combining immobilization and non-weightbearing synergistically promotes MAT accumulation. Additionally, it is unknown whether MAT increase induced by immobilization or non-weightbearing can be reversed through remobilization or reloading. We aimed to address these questions. ACL-reconstructed rats were divided into four groups: no intervention, immobilization, non-weightbearing, or immobilization plus non-weightbearing. Immobilization and non-weightbearing were applied for 2 weeks, after which all rats were allowed to move unrestricted. Intact rats were used as controls. The marrow adiposity and trabecular bone in the proximal tibia were histologically assessed at 2-, 4-, and 12-weeks post-surgery. ACL reconstruction induced MAT accumulation and trabecular bone loss accompanied by increased osteoclastogenesis. Two weeks of immobilization and non-weightbearing after ACL reconstruction individually promoted MAT accumulation, but the combined use of these interventions had a similar impact on MAT accumulation as either of each intervention. Importantly, the increased MAT induced by immobilization or non-weightbearing did not reverse even after remobilization or reloading. Neither immobilization, non-weightbearing, nor both conditions combined after ACL reconstruction further decreased trabecular bone compared to no intervention. These findings suggest no synergistic effect of immobilization and non-weightbearing on MAT accumulation, and MAT accumulation induced by 2 weeks of both immobilization or non-weightbearing did not decrease even after at least 10 weeks of remobilization or reloading. MAT accumulation due to both immobilization and non-weightbearing did not have negative effects on trabecular bone.

Cervicovaginal (CV) microbiota is critical for the well-being of host. We investigated the relationship between the ratio of Lactobacilli (LB) and cocci/coccobacilli (C/CB)-type microbial cells with biofilm formation of CV mixed cultures of women with no inflammation/infection or any epithelial abnormalities in Pap-stained smears Group 1 (G1) corresponds to the flora with LB-type cells alone, whereas G2 corresponds to the LB-dominated flora. G3 contains balanced LB and C/CB cells and G4 is dominated with C/CB. G5 corresponds to a flora with C/CB-type cells alone. Biofilm formation of CV mixed cultures was assessed by crystal violet binding assay and optical density (OD)≥0.8 were defined as biofilm producers. G1 and G3 exist in higher frequencies compared to the other smear groups. However, although the frequency of G5 dominated with C/CB-type cells were the lowest (4%); biofilm formation in that group was observed in the highest frequency (42.9%). The least biofilm formation frequency was observed in G3 smears with balanced flora (1%). Biofilm formation in healthy CV flora increases when there becomes an imbalance between LB and C/CB-type cells and an increase in C/CB-type cells. Our approach may enable early detection of vaginal dysbiosis in healthy flora prone to biofilm-associated CV infections such as bacterial vaginosis (BV).

This study explores the role of irisin and interleukins in parotid tumors by determining the tissue staining intensity of irisin, the salivary and plasma levels of irisin, and the plasma levels of IL-4, IL-6, IL-10 and TNF-alpha in individuals with parotid tumors. Forty-eight patients and forty healthy individuals were included to the study and allocated into four group. Benign Group I (pleomorphic adenoma), Group II (Warthin's tumor), Group III (mucoepidermoid carcinoma) and Group IV (benign parotid control group, healthy control group). Parotid tissue, plasma and saliva samples were collected from each of the patients with parotid tumors, while plasma and saliva samples were collected from the healthy individuals. Normal parotid tissue for histologic evaluation was obtained from unaffected areas in patients with parotid tumors. The levels of irisin in the plasma and saliva were significantly lower in the parotid tumors, while the plasma levels of IL-6 and TNF-alpha were higher in patients with parotid tumors, but no statistically significant difference in IL-4 and IL-10 levels was found. The histopathological intensity of FNDC-5/irisin staining was significantly decreased in the parotid tumor tissues when compared to the benign parotid control group with normal parotid tissue. The low histopathological tissue staining intensity and plasma and salivary levels of irisin suggest that irisin may be a protective protein in parotid tumors.

We have examined some effects of administering vitamin D and extract of common nettle (Urtica dioica) to rats with experimentally induced Crohn's disease (CR). Body weight and colon length were lower in the CR group than in normal controls, whereas scores for histopathologic changes seen in sections stained by the H&E and PAS methods were lower in rats with CR than in those that also received either vitamin D (CRD) or nettle extract (CRI). Strong manganese-superoxide dismutase (Mn-SOD) immunoreactivity was detected in the crypt epithelium of the CR and CRI groups and in the lymphoid tissue of the CRD group. Weak catalase (CAT) immunoreactivity in the crypt epithelium in the CR, CRI, and CRD groups and strong CAT immunoreactivity in the lymphoid tissue in the CR group were also observed. Our results reveal that administering either vitamin D and common nettle extract can have augment Mn-SOD and CAT expression in colon tissues and contribute to alleviation of some complications of experimental Crohn's disease.

Myoepithelial cells (MECs) are known to play an active role in mixed mammary tumors and are found in dogs as well as in humans. The study aimed to assess the morphologic features of epithelial and mesenchymal cells and MECs and investigate their roles in epithelial-mesenchymal transformation in different tumor types in canine mammary tumors. Immunohistochemical staining was performed on 165 specimens from benign mixed tumors (BMT), carcinosarcomas, and simple carcinomas (SC). Double immunohistochemical staining was for the antigen pairs p63/actin, p63/vimentin, p63/CK19, p63/CK8, p63/CK14. BMP6 was demonstrated only by single immunostaining. For BMP6, epithelial cells were positive in BMTs, carcinosarcomas, and SCs. Myoepithelial cells were variably positive for p63, actin, vimentin, and CK14. Epithelial cells were usually positive for CK19 and CK8. MECs include both epithelial and mesenchymal components, so metaplasia in epithelial cells can also form in MECs, and these two cell types should be evaluated together.

Acetylcarvacrol is a semi-synthetic product derived from carvacrol and has known activity against ticks. In vertebrates, the thyroid has been used as a bioindicator in toxicity studies due to its sensitivity to external factors. Thus, the objective of this study was to evaluate the toxic effects of acetylcarvacrol in Wistar rats subjected to repeated dose dermal and oral toxicity tests by means of histopathological analysis of the thyroid. For each test, the rats were divided into 4 groups containing 5 animals. In the topical treatment test, acetylcarvacrol was applied to the trichotomized back of each animal at concentrations of 26, 52 and 104 μL/mL for 21 days. In the oral test, the animals were fed acetylcarvacrol by gavage at concentrations of 26, 52 and 104 μL/mL for 30 days. The control groups were treated only with the vehicles. A significant increase in interstitial tissue vascularization was observed in the group treated topically with the highest concentration of acetylcarvacrol compared to the control. No significant changes were observed between the treatment and control groups in the oral experiment. The comparison between the treated groups and their respective controls also showed no differences in the colloid, the follicle and the follicular cells. The reduced occurrence of changes in this tissue suggests relative safety for use in the control of ticks, although caution is needed when using it at high concentrations or for long periods of time.