Biotechnic & Histochemistry最新文献

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.

Metabolic syndrome (MetS) is a prevalent public health problem. Uric acid (UA) is increased by MetS. We investigated whether administration of UA and 10% fructose (F) would accelerate MetS formation and we also determined the effects of irisin and exercise. We used seven groups of rats. Group 1 (control); group 2 (sham); group 3 (10% F); group 4 (1% UA); group 5 (2% UA); group 6 (10% F + 1% UA); and Group 7, (10% F + 2% UA). After induction of MetS (groups 3 -7), Group 3 was divided into three subgroups: 3A, no further treatment; 3B, irisin treatment; 3C, irisin treatment + exercise. Group 4, 1% UA, which was divided into three subgroups: 4A, no further treatment; 4B, irisin treatment; 4C, Irisin treatment + exercise. Group 5, 2% UA, which was divided into three subgroups: 5A, no further treatment; 5B, irisin treatment; 5C, irisin treatment + exercise. Group 6, 10% F + 1% UA, which was divided into three subgroups: 6A, no further treatment; 6B, irisin treatment; 6C, irisin treatment + exercise. Group 7, 10% F + 2% UA, which was divided into three subgroups: 7A, no further treatment; 7B, irisin treatment; 7C, irisin treatment + exercise., İrisin was administered 10 ng/kg irisin intraperitoneally on Monday, Wednesday, Friday, Sunday each week for 1 month. The exercise animals (in addition to irisin treatment) also were run on a treadmill for 45 min on Monday, Wednesday, Friday, Sunday each week for 1 month. The rats were sacrificed and samples of liver, heart, kidney, pancreas, skeletal muscles and blood were obtained. The amounts of adropin (ADR) and betatrophin in the tissue supernatant and blood were measured using an ELISA method. Immunohistochemistry was used to detect ADR and betatrophin expression in situ in tissue samples. The duration of these experiments varied from 3 and 10 weeks. The order of development of MetS was: group 7, 3 weeks; group 6, 4 weeks; group 5, 6 weeks; group 4, 7 weeks; group 3, 10 weeks. Kidney, liver, heart, pancreas and skeletal muscle tissues are sources of adropin and betatrophin. In these tissues and in the circulation, adropin was decreased significantly, while betatrophin was increased significantly due to MetS; irisin + exercise reversed this situation. We found that the best method for creating a MetS model was F + UA2 supplementation. Our method is rapid and simple. Irisin + exercise was best for preventing MetS.

Coccidiosis is one of the most common infectious diseases in goat farming. The disease causes major economic loss in the world. In this study, we aimed to investigate the activity of heat shock protein 70 in intestine cells of goats with coccidiosis. We used total of twenty-seven goats for this purpose. Gross findings were diarrhoea, cachexia, and dehydration. In the microscopical examination, we observed proliferative enteritis with Eimeria. parasites. Immunohistochemical examinations revealed moderate to severe Hsp70 immunoreactivity in intestines. Considering Hsp70 is a stress protein with anti-apoptotic and immune regulatory features, Hsp70 immunoreactivity attributed to the stress caused by infection and anti-apoptotic activity of the protein along with immune regulatory effects of Hsp70.

Arsenic exposure is associated with numerous morbidities due to dysfunction of various organ systems including the gastrointestinal tract. We investigated the protective effect of grape seed oil (GSO) against sodium arsenite (NaAsO₂)-induced gastric, hepatic and colonic injuries in rats. Twenty-four male Wistar rats were divided into four groups of six as follows: Group A (control) received saline; group B received NaAsO₂ (2.5 mg/kg) orally for 7 days; group C were treated concurrently with NaAsO₂ and GSO (2 ml/kg), while group D received only GSO. Administration of NaAsO₂ induced significant (p < 0.05) increases in alanine aminotransferase (ALT) and aspartate aminotransferase (AST); increased periodic acid Schiff (PAS) staining for mucus and increased goblet cell numbers in the stomach and colon; inflammatory cell infiltration and vascular congestion and alterations in the fecal bacterial flora. GSO supplementation generally promoted a reversal of changes induced by NaAsO₂ towards control levels. Additionally, there was increased immunohistochemically detected expression of colonic B-cell lymphoma-1 (Bcl-2) and cytokeratins AE1/AE3, but reduced expression of mucin 1 (MUC1) and carcinoembryonic antigen (CEA) in NaAsO₂ + GSO and GSO treated rats when compared with the NaAsO₂ group. These results suggest that GSO promoted anti-inflammatory processes in the liver, stomach and colon, as well as opposing apoptosis in the colon, resulting in significant attenuation of damage to these tissues.

The application of most chemical fixatives, such as formalin, in the anatomic pathology laboratory requires safety training and hazardous chemical monitoring due to the toxicity and health risks associated with their use. Consequently, the use of formalin has been banned in most applications in Europe; the primary exception is its use in the histology laboratory in lieu of a suitable and safer alternative. Glyoxal based solutions, several of which are available commercially, are the most promising alternative fixatives, because they are based on a mechanism of fixation similar to that of formalin. Unlike formalin, however, glyoxal based solutions do not dissociate from water and therefore do not require ventilation measures such as a fume hood. A primary barrier to the adoption of commercially available glyoxal based solutions is their low pH, which can produce undesirable morphological and antigenic tissue alterations; however, a recently available neutral pH glyoxal product (glyoxal acid free) (GAF) has been developed to mitigate the challenges of low pH. We compared the morphology and histochemistry among tissues fixed in 10% neutral buffered formalin, a commercially available acidic glyoxal product (Prefer), and GAF. Tissues fixed in formalin and Prefer exhibited similar morphology and staining properties; tissues fixed with 2% GAF exhibited deleterious effects.

In the brains of adult rodents, the cells arising in the subventricular zone of the lateral ventricles maintain the ability to divide when migrating to the olfactory bulb along the rostral migratory stream (RMS). Dividing cells in the RMS are most frequently revealed through immunohistochemical detection of an exogenous marker of proliferation, 5-Bromo-2-deoxyuridine (BrdU), which incorporates into DNA during the S-phase of mitosis. The more recently recognized antigen Ki-67 (also known as Kiel-67 and MKI67), an endogenous protein expressed in nuclei at all stages of mitosis, is also used for proliferation detection. BrdU and Ki-67 are often used as alternative methods, but they have not previously been compared in the RMS. We analyzed the numbers and distribution of cells labeled either with BrdU or Ki-67 within the RMS of adult rats. The first group of animals received a single i.p. dose of BrdU. In the second group, dividing cells were visualized by Ki-67 immunohistochemistry. Some sections from brains of BrdU-treated rats were also immunostained for Ki-67. Labeled cells were counted in the three anatomical parts of the RMS (vertical arm, elbow and horizontal arm) using a method for unbiased estimation of cell density. The distribution of proliferating cells was similar for both markers. Most BrdU and Ki-67 positive cells were located in the vertical arm and in the elbow, but a caudo-rostral reduction in cell divisions was more evident with Ki-67 labeling. The number of Ki-67 positive cells significantly exceeded the number of BrdU positive cells in all parts of the RMS. Our results indicate that BrdU and Ki-67 are not interchangeable markers for evaluation of proliferative activity in the RMS.

Romanowsky staining was an important methodological breakthrough in diagnostic hematology and cytopathology during the late 19^th and early 20^th centuries; it has facilitated for decades the work of biologists, hematologists and pathologists working with blood cells. Despite more than a century of studying Romanowsky staining, no systematic review has been published that explains the chemical processes that produce the "Romanowsky effect" or "Romanowsky-Giemsa effect" (RGE), i.e., a purple coloration arising from the interaction of an azure dye with eosin and not due merely to their simultaneous presence. Our review is an attempt to build a bridge between chemists and biomedical scientists and to summarize the available data on methylene blue (MB) demethylation as well as the related reduction and decomposition of MB to simpler compounds by both light and enzyme systems and microorganisms. To do this, we analyze modern data on the mechanisms of MB demethylation both in the presence of acids and bases and by disproportionation due to the action of light. We also offer an explanation for why the RGE occurs only when azure B, or to a lesser extent, azure A is present by applying experimental and calculated physicochemical parameters including dye-DNA binding constants and electron density distributions in the molecules of these ligands. Finally, we discuss modern techniques for obtaining new varieties of Romanowsky dyes by modifying previously known ones. We hope that our critical literature study will help scientists understand better the chemical and physicochemical processes and mechanisms of cell staining with such dyes.

Silymarin and thymoquinone exert neuroprotective effects, although their combined effects in focal cerebral ischemia/reperfusion (I/R) models are unknown. We compared the effect of silymarin and thymoquinone in an I/R rat model. Wistar rats were divided into five groups: SHAM, REP (I/R), SIR (200 mg/kg silymarin+I/R), TIR (3 mg/kg thymoquinone+I/R), and STIR (200 mg/kg silymarin+3-mg thymoquinone+I/R). The rats underwent bilateral carotid artery occlusion for 30 min and neurological assessments 24 h thereafter. Apoptosis was evaluated using anti-caspase-3 and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) assays. Astrocyte activation was determined using an anti-GFAP antibody. Total antioxidant status (TAS), total oxidant status (TOS), and trimethylamine N-oxide (TMAO) levels were measured. SHAM and REP rats had the lowest and highest neurological scores, respectively (p = 0.001). REP rats showed greater deterioration than SIR, TIR, and STIR rats. SIR, TIR, and STIR rats had fewer TUNEL and caspase-3-positive cells than REP rats (p<0.05). GFAP expression was higher in REP rats (p<0.05) than in SIR, TIR, and STIR rats (p<0.05). SIR and TIR rats showed higher TAS than REP rats (p<0.05). SIR, TIR, and STIR rats had lower TMAO values than REP and SHAM rats (p<0.05). Silymarin/thymoquinone reduces impairment, apoptosis, and astrocyte activation. Combination therapy reduces TMAO levels.