Histopathological and immunohistochemical investigation of the effect of Shilajit in rats with experimental spinal cord injury.

BACKGROUND: This experimental study was designed to investigate the histopathological and immunohistochemical effects of Shilajit in rats with experimentally induced spinal cord injury (SCI). METHODS: The rats were divided into three groups: Control group: The group in which spinal cord damage was created but no drug was administered. Low-dose group: This is the group in which intraperitoneal Shilajit is given at a dose of 150 mg/kg at the 1st h, 1st day, 2nd day, and 3rd day after spinal cord damage was induced. High-dose group: This is the group in which intraperitoneal Shilajit is given at a dose of 250 mg/kg at the 1st h, 1st day, 2nd day, and 3rd day after spinal cord damage was induced. Thin sections taken from the spinal cord after euthanasia were sent for histopathological and immunohistochemical examination. RESULTS: Histopathological examination of the high-dose group showed lower amounts of morphological findings compared to the low-dose group and control group. While a significant CD68 immune reaction was observed in the control group of rats with spinal injury, the positive immune reaction was found to be significantly decreased in the Shilajit-applied groups. CONCLUSION: It is thought that the use of Shilajit in SCI will reduce the effects of secondary damage in SCI and that its administra-tion to such patients will have positive effects on the results.

Exploring of the ameliorative effects of Nerium (Nerium oleander L.) ethanolic flower extract in streptozotocin induced diabetic rats via biochemical, histological and molecular aspects.

BACKGROUND: Nerium oleander L. is ethnopharmacologically used for diabetes. Our aim was to investigate the ameliorative effects of ethanolic Nerium flower extract (NFE) in STZ-induced diabetic rats. METHODS: Seven random groups including control group, NFE group (50 mg/kg), diabetic group, glibenclamide group and NFE treated groups (25 mg/kg, 75 mg/kg, and 225 mg/kg) were composed of forty-nine rats. Blood glucose level, glycated hemoglobin (HbA1c), insulin level, liver damage parameters and lipid profile parameters were investigated. Antioxidant defense system enzyme activities and reduced glutathione (GSH) and malondialdehyde (MDA) contents and immunotoxic and neurotoxic parameters were determined in liver tissue. Additionally, the ameliorative effects of NFE were histopathologically examined in liver. mRNA levels of SLC2A2 gene encoding glucose transporter 2 protein were measured by quantitative real time PCR. RESULTS: NFE caused decrease in glucose level and HbA1c and increase in insulin and C-peptide levels. Additionally, NFE improved liver damage biomarkers and lipid profile parameters in serum. Moreover, lipid peroxidation was prevented and antioxidant enzyme activities in liver were regulated by NFE treatment. Furthermore, anti-immunotoxic and anti-neurotoxic effects of NFE were determined in liver tissue of diabetic rats. Histopathogically, significant liver damages were observed in the diabetic rats. Histopathological changes were decreased partially in the 225 mg/kg NFE treated group. SLC2A2 gene expression in liver of diabetic rats significantly reduced compared to healthy rats and NFE treatment (25 mg/kg) caused increase in gene expression. CONCLUSION: Flower extract of Nerium plant may have an antidiabetic potential due to its high phytochemical content.

Evaluation of the relationship between inflammatory reaction and interleukins in ovine pulmonary adenocarcinomas.

In this study, it was aimed to investigate the association between inflammatory reaction of tumoral microenvironments with interleukin responses in ovine pulmonary adenocarcinomas (OPAs). Material of the study consisted of 26 sheep lung tissue samples being brought to the Pathology Department for routine diagnosis. Cases were collected between years 2009 - 2021; pre-diagnosis was based on clinical symptoms, anamnesis and gross lesion of the lungs. These tissues were designated in two groups as control (n = 6) and OPA (n = 20) groups. Choice of immunohistochemical staining was avidin-biotin peroxidase method. Reverse transcription polymerase chain reaction (RT-PCR) was used to confirm Jaagsiekte sheep retrovirus from paraffin-embedded tissues. On gross examination of OPAs, lesions seen were mostly in the caudal lobes of the lung, 1.00 - 2.00 cm in diameter as gray-white consolidated foci and in microscopic observation, tumor cells showed acinar, papillary or mixed growths. No expressions of interleukin (2 and 8) were observed in the control group. All OPAs cases were positive for interleukins (2 and 8) expressions. A total of eight tissue samples were detected as positives through RT-PCR. In conclusion, in this study, it was determined that interleukin-2 and interleukin-8 were produced from tumor microenvironment elements, especially tumor-associated macrophages, and these interleukins showed pro-inflammatory effects. Interleukins and the inflammatory reaction may promote the development of OPA.

The effect of theranekron on femur fracture healing in an experimental rat model.

OBJECTIVES: The aim of this study was to investigate the radiological, biomechanical, histopathological and immunohistochemical effects of theranekron on fracture healing in an experimental rat model. MATERIALS AND METHODS: Forty-eight male albino Wistar rats were used. Four groups were formed, with 12 rats in each of theranekron groups 1 and 2, and control groups 1 and 2. After a fracture was created in the right femur of the rats included in the study, fixation was performed with an intramedullary Kirschner wire. Theranekron was administered subcutaneously to theranekron groups 1 and 2 at a dose of 0.3 mg/kg on days 0, 5 and 10. After radiographic analysis of the femurs of theranekron group 1 and control group 1 rats at four weeks of the study was performed, both groups were divided into two equal subgroups (six femurs in each group). Histopathological and immunohistochemical examinations were performed in one subgroup and biomechanical examination in the other subgroup. At the end of six weeks, the rats in theranekron group 2 and control group 2 were evaluated after applying the same procedure as in the fourth week. RESULTS: When the mean radiological scores of the theranekron and control groups were compared, a statistically significant difference was found in favor of the theranekron group at four and six weeks (p=0.028 and p=0.006, respectively). At four weeks, statistically significant higher biomechanical forces were obtained in the theranekron group compared to the control group (p=0.030). In the histopathological evaluation, the inflammation value of the control group at four weeks was statistically significantly higher than the theranekron group (p=0.027). The angiogenesis, osteoblast proliferation, and bone formation values of the theranekron group were significantly higher than the control group (p=0.014, p=0.014, and p=0.005, respectively). At six weeks, the bone formation values of the theranekron group were statistically significantly higher than the control group (p=0.021). The difference between the theranekron group and the control group scores of the immunohistochemical evaluation were statistically significantly different at four and six weeks (p=0.006 and p=0.011, respectively). CONCLUSION: Theranekron may play a role in accelerating fracture healing by reducing acute inflammation process in the early period of fracture union, increasing fracture strength, angiogenesis, osteoblast proliferation, and bone formation.

The protective effects of the Lactarius deliciosus and Agrocybe cylindracea mushrooms on histopathology of carbon tetrachloride induced oxidative stress in rats.

We investigated the protective effects of L. deliciosus and A. cylindracea supplementation against carbon tetrachloride (CCI4) induced oxidative stress by measuring levels of adenosine deaminase (ADA) and myeloperoxidase (MPO), and by observing histopathological changes in liver and kidney tissues of rats. We divided 36 rats into six groups: control, CCl4, L. deliciosus, A. cylindracea, CCl4 + L. deliciosus, and CCl4 + A. cylindracea. We found that administration of CCI4, A. cylindracea, and CCl4 + A. cylindracea increased MPO and ADA levels. We observed severe hepato-renal degenerative and necrotic lesions in the CCI4, A. cylindracea and CCl4 + A. cylindracea groups. Severe lesions of the liver and kidney were not observed with A. cylindracea administration. CCI4 induced hepato-renal lesions were ameliorated by L. deliciosus extract supplementation. L. deliciosus could be an important dietary antioxidant for preventing histologic lesions in liver and kidney due to CCI4 induced oxidative stress in rats.

The Protective Role of Urtica dioica Seed Extract Against Azoxymethane-Induced Colon Carcinogenesis in Rats.

The aim of this study was to investigate the protective role of Urtica dioica seed (UDS) extract against azoxymethane (AOM)-induced colon carcinogenesis in rats. Thirty-two male Wistar albino rats were divided into four groups: Control, AOM, AOM + UDS, and UDS. The AOM and AOM + UDS groups were induced by AOM (15 mg/kg body weight) subcutaneously once a week for 10 weeks. AOM + UDS and UDS groups additionally received fed with pellets included 30 ml/kg UDS extract. At the end of the trial, blood and colon tissue samples were taken from the rats following necropsy. The gross and histopathological findings revealed that the administration of UDS extract significantly decreased lesions including aberrant cript foci, adenoma, and adenocarcinoma formation both numerically and dimensionally. Immunohistochemically, slight CEA and COX-2, strong Caspase-3 immune-expressions were detected in the group AOM + UDS compared to AOM group. Biochemical examinations indicated that a markedly increase in the malondialdehyde and fluctuated antioxidant defense system constituents levels such as reduced glutathione, glutathione s-transferase, glutathione peroxidase, superoxide dismutase were restored in AOM + UDS group. These results reveal that the UDS may act as a chemopreventive dietary agent, inducing apoptosis, resulting in a significant reduction of colon carcinogenesis.

Chemopreventive efficacy of juniper berry oil (Juniperus communis L.) on azoxymethane-induced colon carcinogenesis in rat.

The aim of this study was to investigate the chemopreventive effects of juniper berry (JB) oil on azoxymethane (AOM)-induced colon cancer in rats. Thirty-two male Wistar albino rats were allocated into four groups: Control, AOM, AOM + JB, and JB groups. Whereas the control group was fed with standard pellet feed, the AOM and AOM + JB groups were administered of AOM (15 mg/kg body weight) subcutaneously once every 2 weeks for 10 weeks. AOM + JB and JB groups additionally received JB oil (100 µl/kg) orally. At the end of the 16-week experimental period, blood and tissue samples were obtained from the rats following necropsy. The macroscopic findings showed that the application of JB oil significantly decreased adenoma and adenocarcinoma formation both numerically and dimensionally. Immunohistochemically, CEA, COX-2, and Ki-67 immune-expressions decreased, and the immune-expression of caspase-3 increased in AOM + JB treated rats. Additionally, JB oil supplementation ameliorated antioxidant defense systems and lipid peroxidation within the colon tissue of AOM + JB treated rats. These results reveal that the JB oil acted as a chemopreventive dietary agent, inhibiting cell proliferation and COX-2 expression and inducing apoptosis, resulting in a significant reduction in colon tumor formation.

Reduction of hepatorenal and pancreatic damage by Ferula elaeochytris extract in STZ induced diabetic rats.

The therapeutic potential and antioxidant capacity of Ferula elaeochytris extract (FE) in the liver, kidney and pancreas of rats with diabetes induced by streptozotocin (STZ) was assessed using biochemistry, histopathology and immunohistochemistry. Forty adult Wistar albino male rats were divided randomly into five groups of eight rats each. The normal control (NC) group was untreated. The diabetes control (DC) group was treated with STZ to induce diabetes. The diabetes + acarbose group (DAC) was treated with STZ, then with acarbose daily for 28 days. The diabetes + FE (DFE) group was treated with STZ, then FE daily for 28 days. DC rats had inflammatory cell infiltration, hydropic degeneration and necrosis, whereas the DFE rats exhibited nearly normal histology. Insulin immunostaining in the pancreatic beta cells was decreased in the DC group compared to the NC group, whereas the DFE group was similar to the NC group. Many serum biomarkers of damage to liver, kidneys or pancreas were elevated in the DC group compared to the NC group; these biomarkers were decreased in the DFE group. The DC group exhibited increased malondialdehyde levels and decreased levels of the antioxidant defense system constituents compared to the NC group. The level of biomarkers the DFE group was close to the NC group. FE exhibited a protective effect against tissue damage owing to its antioxidant activities and to its ability to effect regeneration of ß-cells in STZ induced diabetic rats.

Comparison of the protective effects of curcumin and caffeic acid phenethyl ester against doxorubicin-induced testicular toxicity.

Whether testicular toxicity is mediated by matrix metalloproteinases (MMPs) is an important question that has not been examined. This study investigated the suppressive effect of curcumin and caffeic acid phenethyl ester (CAPE) on oxidative stress, apoptosis, and whether MMPs mediate doxorubicin (DOX)-induced testicular injury. Male rats were randomly divided into eight groups (n = 8 per group). The groups were as follows: sham, dimethyl sulphoxide (100 µL), DOX (3 mg/kg), CAPE (2.68 mg/kg), curcumin (30 mg/kg), DOX+CAPE (3 mg/kg DOX and 2.68 mg/kg CAPE), DOX+curcumin (3 mg/kg DOX and 30 mg/kg curcumin) and DOX+CAPE+curcumin (3 mg/kg DOX, 2.68 mg/kg CAPE and 30 mg/kg curcumin). Injections were administered daily for 21 days. The oxidative stress, MMPs, proinflammatory cytokines and apoptotic markers in the DOX group were higher than the sham group (p < .05); these measures were lower in the groups treated with CAPE and curcumin together with DOX compared with the DOX group (p < .05). The results showed that MMPs mediated DOX-induced testicular injury, but CAPE and especially curcumin suppressed testis injury and cell apoptosis by suppressing DOX-induced increases in MMPs, oxidative stress and proinflammatory cytokines. However, curcumin exhibited more pronounced effects than CAPE in terms of all studied parameters.

An Immunohistochemical Study on the Presence of Nitric Oxide Synthase Isoforms (nNOS, iNOS, eNOS) in the Spinal Cord and Nodose Ganglion of Rats Receiving Ionising Gamma Radiation to their Liver.

INTRODUCTION: This study determined the presence of nitric oxide synthesis isoforms (nNOS, iNOS, and eNOS) in thoracic spinal cord segments and nodose ganglia of rats with gamma-irradiated livers. MATERIAL AND METHODS: Male rats (n = 32) were divided into equal groups A, B, C, and D. In group A, the controls, no radiation was applied, while groups B, C, and D received 10 Gy of ionising gamma radiation. The rats of group B were euthanized at the end of the first day (d1), those of group C on the second day (d2), and those of group D on the third day (d3). The liver, spinal cord segments, and nodose ganglion tissues were dissected and fixed, and the liver sections were examined histopathologically. The other tissues were observed through a light microscope. RESULTS: Regeneration occurred at the end of d3 in hepatocytes which were radiation-damaged at the end of d1 and d2. On d1, some nNOS-positive staining was found in the neuronal cells of laminae I-III of the spinal cord and in neurons of the nodose ganglion, and on d3, some staining was observed in lamina X of the spinal cord, while none of note was in the nodose ganglion. Dense iNOS-positive staining was seen on d1 in the ependymal cells of the spinal cord and in the glial cells of the nodose ganglion, and on d3, there was still considerable iNOS staining in both tissues. There was clear eNOS-positive staining in the capillary endothelial cells of the spinal cord and light diffuse cytoplasmic staining in the neurons of the nodose ganglion on d1, and on d3, intense eNOS-positive staining was visible in several endothelial cells of the spinal cord, while light nuclear staining was recognised in the neurons of the nodose ganglion. CONCLUSION: The nNOS, iNOS, and eNOS isoforms are activated in the spinal cord and nodose ganglion of rats after ionising radiation insult to the liver.

Use of an antiarrhythmic drug against acute selenium toxicity.

OBJECTIVE: Selenium is an essential trace element. But, selenium may have toxic effects in high doses. There are no proven antidotes or curative treatments for acut selenium toxicity. Treatment involves stopping the exposure and providing supportive care for symptoms. Therefore, it is necessary to find more effective substances in the treatment of selenium toxicity. The aim of this study was to increase the survival rate of animals by supporting the heart with amiodarone and to determine the effect of amiodarone on the pathological, hematological and biochemical parameters in acute selenium intoxication. METHODS: 64 Wistar-Albino rats were divided into four groups. Group I was given only distilled water, Group II was given 18 mg/kg dose of amiodarone, Group III was given 18 mg/kg amiodarone and 10 mg/kg sodium selenite and Group IV was given sodium selenite 10 mg/kg (LD50 dose)orally. RESULTS: 11 of the 16 animals in Group IV died within the first 48 h of drug administration. However, no deaths were observed in the rats in Group III. No hematological changes were observed. Biochemically, CK, CK-MB and LDH levels of Group IV were higher than the other groups on both the 2nd and 10th days. In Groups II and III, this serum level decreased, and vitamin B12 levels increased. In macroscopic inspections of the organs of Groups III and IV, slight paleness was detected. Histopathologically, degenerative changes in tissue were observed, especially in Group IV. CONCLUSION: This study shows that amiodarone application has a reducing effect on selenium toxicity. This was because amiodarone protected the heart by reducing CK and CK-MB levels and increased vitamin B12 levels, which play a role in the synthesis of S-adenosyl methionine that converts selenium into a nontoxic form.

Congenital extraneural hemangioblastoma in a lamb.

A 1-mo-old Ivesi male lamb was presented with 2 large red masses on the skin of the left ear. The tumors were removed using gentle dissection and submitted for histologic evaluation. The tumors consisted of numerous thin-walled capillaries lined by endothelial cells and nests of stromal cells. Immunohistochemically, the endothelial cells were positive for CD45, and the stromal cells were positive for neuron-specific enolase. GFAP-positive cells were occasionally present within the tumor. Endothelial and stromal cells were negative for S100, CD34, CD31, and factor VIII-related antigen. The tumor had strong gross, microscopic, and immunohistochemical similarities with human extraneural hemangioblastoma.

Detection of Bovine Respiratory Syncytial Virus, Pasteurella Multocida, and Mannheimia Haemolytica by Immunohistochemical Method in Naturally-infected Cattle.

INTRODUCTION: The aim of this study was to determine the predisposing effect of bovine respiratory syncytial virus (BRSV) on Pasteurella spp. infection in naturally-induced pneumonia in cattle by immunohistochemical labelling. MATERIAL AND METHODS: Lungs of cattle slaughtered in the slaughterhouse were examined macroscopically, and 100 pneumonic samples were taken. The samples were fixed in 10% neutral formalin and embedded in paraffin by routine methods. Sections 5 µm in thickness were cut. The streptavidin-peroxidase method (ABC) was used to stain the sections for immuno-histochemical examination. RESULTS: BRSV antigens were found in the cytoplasm of epithelial cells of bronchi, bronchioles, and alveoles and within inflammatory cell debris and inflammatory exudate in bronchial lumens. Pasteurella spp. antigens were detected in the cytoplasm of the epithelial cells of bronchi and bronchioles, and in cells in the lumens of bronchi and bronchioles. Eleven cases were positive for only one pathogen (six for BRSV and five for Pasteurella spp.), while 35 cases were positive for 2 pathogens: BRSV plus P. multocida (n = 21) or M. haemolytica (n = 14). CONCLUSION: The presence of high levels of BRSV in dual infections indicates that BSRV may be the main pneumonia-inducing agent and an important predisposing factor for the formation of Pasteurella spp. infections in cattle naturally afflicted with pneumonia.

Protective effects of silymarin on methotrexate-induced damages in rat testes

Abstract The present study aimed to investigate the protective effects of silymarin (SMN), an antioxidant, on methotrexate (MTX)-induced damage in rat testes. Thirty-two Wistar albino rats were divided into four groups (n = 8): control, MTX (20 mg/kg, i.p. on days 1 and 5), SMN (200 mg/kg, orally), and MTX + SMN (20 mg/kg, i.p. on days 1 and 5 and SMN 200 mg/kg orally) groups. At the end of the 6-week trial period, histopathological, immunohistochemical, biochemical, and spermatological analyses were performed on testes tissues. Histopathologically, MTX-induced damage, including depletion of germ cell and loos of spermatozoa, was significantly improved with SMN treatment. Immunohistochemically, the immunoreactivity of glutathione peroxidase 1 (GPx1) and manganese superoxide dismutase 2 (SOD2) were detected more intensely in the MTX + SMN group than in the MTX group. Biochemical examinations revealed that SMN supplementation decreased the lipid peroxidation and increased enzymatic antioxidants in the SMN-treated rats. Spermatologically, significant differences were found in the density, motility, dead-to-live sperm ratio, and abnormal sperm rate in the MTX + SMN group compared to the MTX group. In conclusion, SMN seems to have protective effects as an antioxidant against MTX-induced damage in rat testes.