Investigation of local ACE activity and structural alterations during development of L-NAME-induced hypertension

Tonic basal release of nitric oxide (NO) by vascular endothelial cells controls blood pressure (BP) in the basal state. In the present study, we showed how serum and local angiotensin converting enzyme (ACE) alters during development of hypertension in chronic nitric oxide synthase blockade, a non-renin-dependent model of hypertension. Four experiments were performed in which animals were given N�-nitro-L-arginine methyl ester (L-NAME) (50 mg kg-1) for 2, 4, 8 and 12 weeks. The control group rats received tap water. The ACE activity in serum, aorta, heart, kidney and lung was analyzed by high performance liquid chromatography (HPLC) and the structural change in aorta was investigated by measurement of cross-sectional area (CSA). Significant elevation of systolic blood pressure developed in chronically NO-blocked rats compared to controls. These results indicated that ACE activity in aortae and heart was gradually increased during development of hypertension and was more pronounced at higher blood pressure. Furthermore, there was a positive correlation between aortic cross-sectional area and elevation of blood pressure. These observations highlight the importance of the local ACE particularly in organs regulating hypertension (aorta and heart) during development of L-NAME-induced hypertension

Study of correlation between systolic blood pressure and Angiotensin Converting Enzyme (ACE) activity in STZ induced diabetic rats

Background: Diabetes mellitus is a worldwide health problem affecting 1-2 of the population. It is responsible for numerous morbidity and mortality consequences due to vascular events such as hypertension, nephropathy and retinopathy. The precise mechanism of hyperglycemia-induced vascular damage is not clearly known. Alteration in Rennin Angiotensin-Aldosterone System (RAAS) and increased Angiotensin Converting Enzyme (ACE) activity is known to be involved in pathogenesis of vascular disorders. This study sought to investigate correlation between systolic blood pressure and ACE activity in STZ induced diabetic rats. Methods: Two groups of 8 male Sprauge Dawely rats including control (C) and diabetic group (D) were used in this study. Diabetes induced by injection of 60 mg/kg STZ intraperitoneally. Blood pressure was measured using tail cuff method. ACE activity was determined by HPLC method. Results: At the end of study (four weeks after induction of diabetes) systolic blood pressure increased significantly in D group compared to control rats. ACE activity was increased in aorta, heart, lung and serum of D group which this increment was more pronounced in aorta and heart. Renal ACE activity reduced significantly in this group compared to control. Conclusion: It is concluded that increased ACE activity particularly cardiovascular ACE, could be involved in the diabetes induced hypertension and vasculopathy

The effect of L-carnitine on serum nitric oxide level and angiotensin converting enzyme activity in STZ - Induced diabetic and normal rats

Background: Cardiovascular disease including hypertension are complications of long-standing diabetes. A few Studies had shown the positive effects of L-carnitine on hypertension. In this study, the possible effects of L-carnitne on nitric oxide (NO) levels and angiotensin-converting enzyme (ACE) activity in serum as well as systolic blood pressure (SBP) in diabetic and normal rats were studied. Methods: In this study forty rats were used in four groups including non-treated control (C), L-carnitne treated control (CT), diabetic (D) and L-carnitne treated diabetic (DT). Diabetes was induced in rats by injection of stereptozotosin. Both of C & D groups had a free access to food and water and CT & DT groups were received daily dose of L-carnation in drinking water. At the end of 12 weeks SBP, serum NO and ACE activity were measured. Results: Systolic blood pressure was significantly decreased in DT group compared to D group. Serum ACE activity was also significantly decreased in DT group compared to D group and the serum NO levels were significantly increased in DT & CT groups compared to D & C groups respectively. Conclusion: Finally it could be concluded that L-carnitine may reduce SBP in diabetic rats via elevation of serum NO levels and reduction of serum ACE activity

Factors Affecting Incidence of Uterine Torsion in Egyptian Buffaloes (Bubalus bubalis) and its Response for Rolling

Objective: The present work was planned to study the incidence of uterine torsion in Egyptian buffaloes as well as its prognosis after mechanical treatment.Methods: A total number of 35 buffaloes of different ages, parities and stage of pregnancy with complain of colic and anorexia were included in the present study. These animals were examined rectally to detect stage of pregnancy, degree, direction, duration, location of uterine torsion. Mechanical handling of torsion cases and prognosis of recovered cases was performed. After birth, sex of new born was also recorded.Results: The torsion of uterus mostly occurred in pleuriparous buffaloes (100%) in right side (85.7%) during second half of pregnancy (100%). Most cases were postcervical (85.7%) during the early stage (7 and 8 months). Torsion also may occur during labour (17.4%) where the cervix was dilated after correction. Most cases of uterine torsion occur in stabled animals for long period (97%). The uterine torsion associated with male calves (65.7%) as well as with large size calves (40-50kg) and majority of the calves were in normal position. The first foetal sac expelled in majority of cases was amniotic sac (97%) with few cases of placental retention. Uterine torsion with short duration (1-6 h) usually needs 1-2 rolls only for correction and usually takes 0-6h (100%) for cervical dilatation. Occurrence of uterine torsion for above three days usually did not respond for rolling.Conclusion: Uterine torsion usually occurs in old stabled animal with good BCS during green season at last month of pregnancy. The right sever and post cervical torsion were the most common type of torsion. Rapid diagnosis of uterine torsion within 6 h usually results in good prognosis even in severe cases

Study of high glucose-induced apoptosis in PC12 cells: Role of bax protein

Hyperglycemia, which occurs under the diabetic condition, induces serious diabetic complications. Diabetic neuropathies, affecting the autonomic, sensory, and motor peripheral nervous system, are among the most frequent complications of diabetes. Little is known about the direct toxic effect of high glucose concentrations on neuronal cells. Therefore in the present study, glucose-induced toxicity was studied in PC12 cells as an in vitro cellular model for diabetic neuropathy using the MTT assay. The possible role of apoptosis was also investigated in this toxicity. The result showed that a 3-fold increase in optimum glucose concentration for PC12 cells (13.5 mg/ml) significantly reduced cell viability after 48 h. In Western blot analysis, the ratio of Bax/Bcl-2 protein expression in cells treated with high glucose was significantly increased compared to controls. Additionally high glucose could induce a DNA ladder pattern in PC12 cells, a hallmark of apoptosis indicating nuclear fragmentation. From our present results, it may be concluded that high glucose can cause PC12 cell death, in which apoptosis plays an important role possibly by the mitochondrial pathway through higher expression of Bax proapoptotic protein. ©2007 The Japanese Pharmacological Society

Deferoxamine preconditioning to restore impaired HIF-1α-mediated angiogenic mechanisms in adipose-derived stem cells from STZ-induced type 1 diabetic rats

Objectives: Both excessive and insufficient angiogenesis are associated with progression of diabetic complications, of which poor angiogenesis is an important feature. Currently, adipose-derived stem cells (ADSCs) are considered to be a promising source to aid therapeutic neovascularization. However, functionality of these cells is impaired by diabetes which can result from a defect in hypoxia-inducible factor-1 (HIF-1), a key mediator involved in neovascularization. In the current study, we sought to explore effectiveness of pharmacological priming with deferoxamine (DFO) as a hypoxia mimetic agent, to restore the compromised angiogenic pathway, with the aid of ADSCs derived from streptozotocin (STZ)-induced type 1 diabetic rats ('diabetic ADSCs'). Materials and methods: Diabetic ADSCs were treated with DFO and compared to normal and non-treated diabetic ADSCs for expression of HIF-1α, VEGF, FGF-2 and SDF-1, at mRNA and protein levels, using qRT-PCR, western blotting and ELISA assay. Activity of matrix metalloproteinases -2 and -9 were measured using a gelatin zymography assay. Angiogenic potential of conditioned media derived from normal, DFO-treated and non-treated diabetic ADSCs were determined by in vitro (in HUVECs) and in vivo experiments including scratch assay, three-dimensional tube formation testing and surgical wound healing models. Results: DFO remarkably enhanced expression of noted genes by mRNA and protein levels and restored activity of matrix metalloproteinases -2 and -9. Compromised angiogenic potential of conditioned medium derived from diabetic ADSCs was restored by DFO both in vitro and in vivo experiments. Conclusion: DFO preconditioning restored neovascularization potential of ADSCs derived from diabetic rats by affecting the HIF-1α pathway. © 2015 John Wiley & Sons Ltd

Glioma Migration Through the Corpus Callosum and the Brainstem Detected by Diffusion and Magnetic Resonance Imaging: Initial Findings

Purpose: Glioma cell infiltration, in which the glioma tumor cells spread long distances from the primary location using white matter (WM) or blood vessels, is known as a significant challenge for surgery or localized chemotherapy and radiation therapy. Following the World Health Organization (WHO), the glioma grading system ranges from stages I to IV, in which lower-grade gliomas represent benign tumors, and higher grade gliomas are considered the most malignant. Materials and Methods: We gathered magnetic resonance imaging (MRI) and diffusion tensor imaging (DTI) data for seven patients with right precentral gyrus-located tumors and six age- and sex-matched healthy subjects for analysis. Tract-Based Spatial Statistics (TBSS) was utilized to evaluate whole-brain WM implication due to probable tumor infiltration. Also, along-tract statistics were used in order to trace the implicated WM tracts. Finally, for cortical evaluation of probable tumor cell migration, voxel-based morphometry (VBM) was utilized, which allowed us to do whole-brain cortical estimation. Results: The TBSS results revealed significantly higher fractional anisotropy (FA) and lower mean diffusivity (MD) in the left side superior corona radiata. Also, higher FA was observed in the right corticostriatal tract. Along-tract statistics were also compiled on the corpus callosum (CC), which is anatomically known as a hub between hemispheres. The body of the CC, which connected with the superior corona radiata anatomically, showed significantly higher FA values relative to healthy subjects, which are in line with the TBSS results. Consistent with these results, whole-brain gray matter changes were analyzed via VBM, which showed significant hypertrophy of both sides of the brainstem. Conclusion: In future investigations, focusing on the genetic basis of the glioma patients in line with imaging studies on a larger sample size, which is known as genetics imaging, would be a suitable approach for tracing this process. © Copyright © 2020 Sharifi, Pajavand, Nateghinia, Meybodi and Hasooni

Case Report: Chemotherapy Indication in a Case of Neurofibromatosis Type 1 Presenting Optic Pathway Glioma: A One-Year Clinical Case Study Using Differential Tractography Approach

Neurofibromatosis type 1 (NF1) is associated with peripheral and central nervous system tumors. It is noteworthy that the regions in which these tumors frequently arise are the optic pathways (OPs) and the brainstem. Thus, we decided to trace the procedure of diffusion Magnetic Resonance Imaging (dMRI) alterations along with Short-Wavelength Automated Perimetry (SWAP) examinations of the OPs after surgery and chemotherapy over 1 year, which enabled us to evaluate chemotherapy's efficacy in an NF1 patient with an OP tumor. In this study, a 25-year-old woman with NF1 and left optic radiation (OR) glioma underwent surgery to remove the glioma. Immunohistochemistry (IHC) revealed a Pilocytic Astrocytoma (PA) WHO grade I. Post-operation chemotherapy done using nine treatment cycles of administering Temozolomide (TMZ) for 5 days every 4 weeks. Applying the region of interest (ROI) differential tractography method and SWAP four times every 3 months allowed us to follow the patient's visual acuity alterations longitudinally. The differential deterministic tractography method and statistical analyses enabled us to discover the white matter (WM) tracts anisotropy alterations over time. Furthermore, statistical analyses on the SWAP results along time illustrated possible alterations in visual acuity. Then, we could compare and associate the findings with the SWAP examinations and patient symptoms longitudinally. Statistical analyses of SWAP tests revealed a significant improvement in visual fields, and longitudinal differential tractography showed myelination and dense axonal packing in the left OR after 1 year of treatment. In this study, we examined an old hypothesis suggesting that chemotherapy is more effective than radiotherapy for NF1 patients with OP gliomas (OPGs) because of the radiation side effects on the visual field, cognition, and cerebrovascular complications. Our longitudinal clinical case study involving dMRI and SWAP on a single NF1-OPG patient showed that chemotherapy did not suppress the OP myelination over time. However, it should be noted that this is a clinical case study, and, therefore, the generalization of results is limited. Future investigations might focus on genetic-based imaging, particularly in more cases. Further, meta-analyses are recommended for giving a proper Field Of View (FOV) to researchers as a subtle clue regarding precision medicine. © Copyright © 2021 Pajavand, Sharifi, Anvari, Bidari-Zerehpoosh, Shamsi, Nateghinia and Meybodi

Expression and function of c1orf132 long-noncoding rna in breast cancer cell lines and tissues

miR-29b2 and miR-29c play a suppressive role in breast cancer progression. C1orf132 (also named MIR29B2CHG) is the host gene for generating both microRNAs. However, the region also expresses longer transcripts with unknown functions. We employed bioinformatics and experimental approaches to decipher C1orf132 expression and function in breast cancer tissues. We also used the CRISPR/Cas9 technique to excise a predicted C1orf132 distal promoter and followed the behavior of the edited cells by real-time PCR, flow cytometry, migration assay, and RNA-seq techniques. We observed that C1orf132 long transcript is significantly downregulated in triple-negative breast cancer. We also identified a promoter for the longer transcripts of C1orf132 whose functionality was demonstrated by transfecting MCF7 cells with a C1orf132 promoter-GFP construct. Knocking-out the promoter by means of CRISPR/Cas9 revealed no alterations in the expression of the neighboring genes CD46 and CD34, while the expression of miR-29c was reduced by half. Furthermore, the promoter knockout elevated the migration ability of the edited cells. RNA sequencing revealed many up-and downregulated genes involved in various cellular pathways, including epithelial to mesenchymal transition and mammary gland development pathways. Altogether, we are reporting here the existence of an additional/distal promoter with an enhancer effect on miR-29 generation and an inhibitory effect on cell migration. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Investigating the Impact of Collagen-Chitosan Derived from Scomberomorus Guttatus and Shrimp Skin on Second-Degree Burn in Rats Model

Background: The present study focused on burning as one of the main causes of mortality with detrimental economic and social effects in the world. The purpose of this study was to investigate the impact of collagen-chitosan gel extracted from Scomberomorus guttatus and shrimp skin in the treatment of second degree burn healing among rats. Materials & method: To fulfill the purpose of the study, chitosan and collagen were extracted respectively from shrimp and Scomberomorus guttatus skin waste by the acid-based method and were evaluated by using Pico Tag, SDS-PAGE. The burn wound healing efficiency of marine collagen-chitosan gel was examined in vivo using rats. Three different ratios of collagen and chitosan blend (Col-CH, 1:3, 1:1 and 3:1) were prepared to obtain the most effective Col-CH gel for burn wound healing and were compared to the animals treated with silver sulfadiazine ointment. Healing burn wound was studied by measuring wound surface area with Image J and histopathologic examination was carried out based on the mean of epithelialization, fibroblastic cells, acute and chronic inflammatory cells, angiogenesis, structure collagen and the amount of collagen on days 15 and 25 post-burn. Results: The results of SDS-PAGE indicated that the extracted collagen was type I and it was composed of two α (α1 and α2) chains. Amino acid analysis showed a much higher glaycin content in extracted collagen which amounted to one-third of the total amino. The wound surface measurement showed a significant reduction in wound size in the group treated with Col-CH (3:1) compared to silver-sulfadiazine treated group on 15th and 25th days. Histopathological findings represented a high score in epithelialization, collagen, collagen structure, fibroblast cell and a decrease in inflammatory cells infiltration in Col-CH (3:1) treated group on 25th day. The most obvious finding of the present study is that chitosan-collagen gel (3:1) represented a better efficacy compared to sulfadiazine in burn wound healing on day 25 post-burn. © 2021 The Japanese Society for Regenerative Medicin