The effect of ouabain on mitochondrial DNA damage in HepG2 cell lines

Our purpose in this study is to analyze mitochondrial DNA (mtDNA) lesion frequencies and mtDNA⁴⁹⁷⁷ deletion in HepG2 cells to examine the effects of ouabain on mtDNA. HepG2 cells were treated with 0.75, 7.5, 75, and 750 nM of ouabain for 24?h in the presence and absence of 10?mM 2-deoxyglucose (2-DG). The frequency of mtDNA⁴⁹⁷⁷ deletions and mitochondrial lesions were determined by real-time polymerase chain reaction. A ??1.2-fold change or greater was considered significant. Ouabain doses of 750, 75, and 7.5 nM alone increased the frequency of mtDNA⁴⁹⁷⁷ deletions 1.39, 1.92, and 1.44 times, respectively. The 750 and 75 nM ouabain doses combined with 2-DG increased the mtDNA⁴⁹⁷⁷ deletion frequency 4.94 and 1.57 times, respectively. The 750 and 75 nM ouabain doses alone increased the mtDNA lesion frequency 2.5 and 1.5 times, respectively. The 750 nM ouabain dose combined with 2-DG increased the mtDNA lesion frequency 2.28 times. The 7.5 nM ouabain dose alone and combined with 2-DG decreased the mtDNA lesion frequency 0.67 and 0.45 times, respectively. Ouabain alone and when combined with 2-DG increases mtDNA lesion and mtDNA⁴⁹⁷⁷ deletion frequencies. This supports the thesis that ouabain creates oxidative stress and induces DNA damage and apoptosis.     

Matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase (TIMP-1) in patients with relapsing-remitting multiple sclerosis treated with interferon beta

OBJECTIVES: Matrix metalloproteinases (MMPs), particularly MMP-9, facilitate T-cell migration into the central nervous system. They play a key role in the disruption of the blood-brain barrier (BBB) and thus in the pathogenesis of multiple sclerosis. Interferon beta's (IFNbeta) ability to alter the balance between MMP-9 and MMP-9s natural inhibitor, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), may play a role in stabilizing the BBB. The aim of this study, was to evaluate serum MMP-9 and TIMP-1 and cerebrospinal fluid (CSF) TIMP-1 levels in patients with relapsing-remitting multiple sclerosis (RRMS) treated with IFNbeta-1a. PATIENTS AND METHODS: Blood and CSF samples from 14 patients with RRMS before and 6 months after IFNbeta therapy and 14 age and sex-matched controls were obtained. Levels of MMP-9 and TIMP-1 were measured using ELISA. RESULTS: Before treatment, patients with MS had higher levels of serum MMP-9 and a higher MMP-9/TIMP-1 ratio than the controls. Although serum levels of TIMP-1 were lower in RRMS patients than in the controls, the differences did not reach statistical significance. CSF levels of TIMP-1 were significantly lower in RRMS patients. In the sixth month of IFNbeta therapy serum MMP-9 and the MMP-9/TIMP-1 ratio were significantly decreased, whereas the changes in serum TIMP-1 were not statistically significant. There was a significant increase in CSF TIMP-1 levels in the sixth month of IFNbeta therapy. CONCLUSIONS: Our result shows that RRMS patients have an impaired MMP-9 and TIMP-1 balance, and that 6 months of IFNbeta therapy is beneficial in restoring this balance.     

Temporary cavernosal-cephalic vein shunt in low-flow priapism treatment

Background

Surgical shunting might be considered as the only option in the treatment of extended ischemic priapism that does not respond to aspiration and medication.

Objective

A modified, simple, minimally invasive, and easily applicable artificial cavernosal–venous shunt technique for treating priapism is described in this report.

Design, setting, and participants

A total of 15 patients with extended and nonresponsive low-flow priapism were treated with this technique between January 1998 and February 2007.

Intervention

When the conservative treatment of low-flow priapism does not yield the expected results, then the temporary cavernosal–cephalic vein shunt should be applied. The standard equipment required for this modified technique includes three angiocaths, two shorn blood serum sets, and saline solution with heparin. The blood in the cavernosa and the saline solution infusion are incorporated into the systemic circulation with the aid of serum sets and angiocaths.

Measurements

Priapism duration, history, causes, its relation with sexual stimulation, pain, and any prior management of priapism were assessed in all patients. A complete blood count and blood gases assessment were conducted in corporal aspirates, and duplex penile ultrasonography was performed, which showed attenuated blood flow in the cavernosal artery.

Results and limitations

This technique was applied in 15 patients. Complete detumescence was achieved in 13. In the two cases in whom the technique did not yield the expected results, there was a need for a sapheno–cavernosal shunt. These patients later complained of erectile dysfunction and penile pain that continued for 6 mo. Of the 13 patients in whom detumescence was achieved, 3 reported erectile dysfunction according to International Index of Erectile Function (IIEF) scores at the 12-mo follow-up.

Conclusions

Artificial cavernosal–cephalic vein shunt in the treatment of priapism is simple, safe, effective, easily applicable, and warrants primary consideration when the second-line treatment of priapism is initiated.     

Gross anatomy and arterial vascularization of the tympanic cavity and osseous labyrinth in mid‐gestational bovine fetuses

This study aims to determine morphological features of certain aural formations, varietal characteristics, and arterial supply in fetal development period in cattle. For this purpose, ears of 10 bovine fetuses in mid‐gestation were evaluated. Organ morphology and vascularization in prenatal life were investigated by using corrosion cast technique. It was observed that some aspects of osseous formation and vascular organization in middle and inner ears were not developed completely in the first half of gestation; in addition, cochlea did not its snail‐like structure yet, lateral semicircular canal was rather low compared to others and auditory ossicles did not take its final shape. The feeding blood vessels of inner ear were found to demonstrate three different distribution patterns, whereas feeding pattern in middle ear was very similar in many specimens. The presence of stapedial artery was also identified. Differences and similarities with other some species were assessed in terms of both general morphological structure and vascular organization. From this regard, it is thought that this study will constitute a comparative model for both humans and other species and provide morphological contributions since there is not sufficient literature on species‐specific ear morphology in the field of veterinary anatomy in contrast to the abundance of studies on humans. Anat Rec, 2010. © 2010 Wiley‐Liss, Inc.     

Primary tumor cells resected from cancer patients and decorated with a novel form of CD80 protein serve as effective antigen-presenting cells for the induction of autologous T cell immune responses ex vivo

Vaccination with autologous tumor cells genetically modified to express costimulatory molecules has shown utility for cancer immunotherapy in preclinical and limited clinical settings. Given the complicated nature of gene therapy, a practical alternative approach has been designed that relies on modification of the cell membrane with biotin and its "decoration" with a chimeric protein composed of the functional portion of human CD80 and core streptavidin (CD80-SA). We tested whether primary tumor cells resected from cancer patients can be decorated with CD80-SA and whether such cells serve as antigen-presenting cells (APCs) to generate autologous T cell responses ex vivo. Tumors and peripheral blood lymphocytes (PBLs) were collected from 14 lung, 9 colon, and 2 breast "treatment-naive" cancer patients presenting various clinical stages of the disease. Tumors were mechanically processed, irradiated, decorated with CD80-SA or control streptavidin (SA) protein, and used as APCs in ex vivo autologous T cell-proliferative and cytotoxicity assays. All tumor samples were modified with CD80-SA, albeit with various degrees of decoration ranging from 21.8 to 100%. CD80- SA-decorated cells generated significant proliferative responses in autologous T cells from 9 of 16 evaluable patients (p < 0.05). Proliferative responses were CD80-SA specific and heterogeneous, with stimulation indices ranging from 0.25 to 45. In 15 of 15 evaluable patients, CD80-SA-specific cytotoxic T cell responses against autologous tumors were generated, 11 of which were significant, with specific killing ranging from 5 to 70%. Taken together, these data demonstrate that primary tumor cells can be effectively decorated with CD80-SA and that such cells serve as APCs to induce autologous antitumor T cell responses.     

Infrared Thermographic Comparison of Temperature Increases on the Root Surface during Dowel Space Preparations Using Circular versus Oval Fiber Dowel Systems

Purpose : The purpose was to evaluate temperature increases during dowel space preparations with oval and circular fiber dowel systems. Materials and Methods : This study included 42 single‐rooted human mandibular premolars. Roots were scanned with cone‐beam computerized tomography (CBCT) to determine the ovoid root canal morphology. Root canals were treated with Ni‐Ti rotary instruments and obturated. A second CBCT was taken to determine the thinnest dentin thickness of each root. Roots were randomly divided into two groups (n = 21) according to the fiber dowel system used: group 1, circular fiber dowel system (D.T. Light‐Post); group 2, oval fiber dowel system (Ellipson Post). Dowel spaces were prepared using a circular fiber dowel drill and a diamond‐coated ultrasonic tip with an oval section under water cooling until 9 mm dowel spaces were obtained. Temperature changes were recorded from the thinnest root surfaces using a FLIR E60 thermal imaging camera. Results : Temperature increases were significantly greater with the circular fiber dowel system than with the oval fiber dowel system (p < 0.05). Conclusions : Although both dowel systems generated high temperature increases on root surfaces, the relatively lower temperature increase associated with the use of oval fiber dowels in ovoid canals makes it preferable to the use of circular fiber dowels.