Oxidative protein damage parameters in plasma in chronic experimental diabetes in rats

An increase in oxidative stress may contribute to the development of oxidative protein damage (OPD) in the streptozotocin-diabetic rat. To show the effect of hyperglycemia in promoting OPD, we determined protein carbonyl (PCO), nitrotyrosine (NT), total thiol (T-SH) and advanced oxidation protein product (AOPP) levels as markers of OPD, and lipid hydroperoxide (LHP) levels as a marker of lipid peroxidation in plasma of acute and chronic diabetic male Sprague-Dawley rats and their controls. The levels of the studied markers, except NT, were determined by colorimetric methods. NT levels were measured by ELISA. Plasma PCO and AOPP levels of chronic diabetic rats were increased significantly compared with those of both acute diabetic rats and the controls. Plasma NT levels of the three groups were not different. Plasma T-SH levels of acute diabetics were increased significantly compared with those of the controls while T-SH increase in the chronic diabetics was not significant. Plasma LHP levels were increased significantly in the chronic diabetic rats compared with those of the controls. The increase in plasma PCO, AOPP, LHP levels in chronic but not in acute diabetic rats may be indicating that persistence of hyperglycemia is involved in the evolution of OPD while plasma NT levels do not seem to reflect OPD in diabetes.     

Estimation of basic uncertainties in clinical analysis.

Clinical analyses have a vital importance in human health. Thus, it is necessary to have reliable analytical information which depends on a good assessment of accuracy and this can be obtained by minimization of uncertainty values. The uncertainty values are complex and arise from different sources such as sample, method, instrumentation and data processing. Some of these components may have been evaluated from the distribution of results of series of measurements and can be characterized by standard deviations. In our laboratory, for example the balance calibration uncertainty for 1 g was found to be 0.00414 g. The uncertainties for the volume of solution contained in a 100 mL volumetric flask and a 0.5 mL glass-pipette were calculated to be 0.0843 mL and 0.0071 mL respectively and etc.     

Water evaporation rates from a model of stratum corneum lipids

Water evaporation rates were measured from thin samples of a model layered structure of stratum corneum lipids with 32% water. A model with only free fatty acids present gave the lowest evaporation rates, while a model with only oleic acid gave values approximately 50% higher. Using the total lipid spectrum of stratum corneum gave a structure with an evaporation rate between those mentioned above. The diffusion coefficients calculated were one to two orders of magnitude higher than those of stratum corneum, reflecting the influence of the proteinous part of the latter structure. A sample with only saturated fatty acids gave extremely high evaporation rates; in fact, they were of the same magnitude as those from an unprotected water surface.     

Cumene hydroperoxide induced changes in oxidation–reduction potential in fresh and frozen seminal ejaculates

Oxidation–reduction potential (ORP) is a newer integrated measure of the balance between total oxidants (reactive oxygen species—ROS) and reductants (antioxidants) that reflects oxidative stress in a biological system. This study measures ORP and evaluates the effect of exogenous induction of oxidative stress by cumene hydroperoxide (CH) on ORP in fresh and frozen semen using the MiOXSYS Analyzer. Semen samples from healthy donors (n = 20) were collected and evaluated for sperm parameters. All samples were then flash‐frozen at ?80°C. Oxidative stress was induced by CH (5 and 50 μmoles/ml). Static ORP (sORP—(mV/10⁶ sperm/ml) and capacity ORP (cORP—μC/10⁶ sperm/ml) were measured in all samples before and after freezing. All values are reported as mean ± SEM. Both 5 and 50 μmoles/ml of CH resulted in a significant decline in per cent motility compared to control in pre‐freeze semen samples. The increase in both pre‐freeze and post‐thaw semen samples for sORP was higher in the controls than with 50 μmoles/ml of CH. The change from pre‐freeze to post‐thaw cORP was comparable. The system is a simple, sensitive and portable tool to measure the seminal ORP and its dynamic impact on sperm parameters in both fresh and frozen semen specimens.     

Arginine-rich cell-penetrating peptide dramatically enhances AMO-mediated ATM aberrant splicing correction and enables delivery to brain and cerebellum

Antisense morpholino oligonucleotides (AMOs) can reprogram pre-mRNA splicing by complementary binding to a target site and regulating splice site selection, thereby offering a potential therapeutic tool for genetic disorders. However, the application of this technology into a clinical scenario has been limited by the low correction efficiency in vivo and inability of AMOs to efficiently cross the blood brain barrier and target brain cells when applied to neurogenetic disorders such as ataxia-telangiecatasia (A-T). We previously used AMOs to correct subtypes of ATM splicing mutations in A-T cells; AMOs restored up to 20% of the ATM protein and corrected the A-T cellular phenotype. In this study, we demonstrate that an arginine-rich cell-penetrating peptide, (RXRRBR)(2)XB, dramatically improved ATM splicing correction efficiency when conjugated with AMOs, and almost fully corrected aberrant splicing. The restored ATM protein was close to normal levels in cells with homozygous splicing mutations, and a gene dose effect was observed in cells with heterozygous mutations. A significant amount of the ATM protein was still detected 21 days after a single 5 μm treatment. Systemic administration of an fluorescein isothiocyanate-labeled (RXRRBR)(2)XB-AMO in mice showed efficient uptake in the brain. Fluorescence was evident in Purkinje cells after a single intravenous injection of 60 mg/kg. Furthermore, multiple injections significantly increased uptake in all areas of the brain, notably in cerebellum and Purkinje cells, and showed no apparent signs of toxicity. Taken together, these results highlight the therapeutic potential of (RXRRBR)(2)XB-AMOs in A-T and other neurogenetic disorders.     

Venous thromboembolism in patients hospitalized with nephrotic syndrome

Purpose

Whether pulmonary embolism in patients with the nephrotic syndrome is caused by deep venous thrombosis or renal vein thrombosis is controversial. To determine which is the likely cause of pulmonary embolism in patients with the nephrotic syndrome, we investigated data from the National Hospital Discharge Survey.

Methods

The number of patients discharged from nonfederal short-stay hospitals in the United States with a diagnostic code of nephrotic syndrome, deep venous thrombosis, renal vein thrombosis, and pulmonary embolism was obtained using ICD-9-M (International Classification of Diseases, Ninth Revision, Clinical Modification) codes.

Results

From 1979 to 2005, 925,000 patients were discharged from hospitals with the nephrotic syndrome and 898,253,000 patients did not have the nephrotic syndrome. With the nephrotic syndrome, 5000 (0.5%) had pulmonary embolism, 14,000 (1.5%) had deep venous thrombosis, and fewer than 5000 had renal vein thrombosis. The relative risk of pulmonary embolism comparing patients with the nephrotic syndrome to those who did not have it was 1.39, and the relative risk of deep venous thrombosis was 1.72. Among patients aged 18-39 years, the relative risk of deep venous thrombosis was 6.81. From 1991-2005, after venous ultrasound was generally available, the relative risk of deep venous thrombosis (all ages) was 1.77.

Conclusion

The nephrotic syndrome is a risk factor for venous thromboembolism. This is strikingly apparent in young adults. Renal vein thrombosis was uncommon. Therefore, pulmonary embolism, if it occurs, is likely to be due to deep venous thrombosis and not renal vein thrombosis.     

The efficacy of paracetamol in the treatment of ankle sprains in comparison with diclofenac sodium

OBJECTIVE: To assess the efficacy of paracetamol in comparison with diclofenac sodium. METHODS: Between February - November 2006, a prospective, double blinded, parallel group study of 100 patients suffering from first or second degree lateral ankle sprain within 48-hours of admission in Tepecik Education and Research Hospital, Izmir, Turkey. Patients with bilateral injury, ipsilateral knee injury, third degree sprain, previous sprain within 6 months, and ankle pain less than 45 according to visual analogue score (VAS) were excluded. Patients rated pain on a 100 VAS, representing 0 no pain, 100 maximal pain. After enrollment, patients were randomized (1:1) with diclofenac sodium 150 mg/day or paracetamol 1500 mg/day for 5 days. Clinical assessments were carried out at baseline; on second, tenth days, and sixth week (end of study). In each visit, VAS and adverse effects of medication were questioned. RESULTS: The mean VAS of the diclofenac group was 81 and 82.3 with paracetamol group at the first visit. These scores decreased to 20.7, 9.9, 4.6 in diclofenac group and 11.9, 6.3, 3 in paracetamol group at the second, tenth days and last examination. Similar reductions in pain were observed at the end of study (p>0.05) in both groups. However, cases treated by paracetamol group showed accelerated decrease in VAS at day 2 and 10 in comparison with diclofenac group (p<0.05). Of the ankle range of motion, there was a similar improvement in both groups (39.6 degrees, 37.5 degrees) (p>0.05). The incidence of gastrointestinal adverse effects on diclofenac group was much more than the paracetamol group, however, there was no significant difference (p>0.05). CONCLUSION: It was concluded that diclofenac sodium and paracetamol are effective and well tolerated as a short term treatment alternatives for acute ankle injuries.     

Results of total knee arthroplasty in patients with gonarthrosis resulting from the delayed diagnosis of PVNS: a case series

Pigmented villonodular synovitis (PVNS) is a benign tumor that affects synovial lined joints, tendon sheaths and bursae. It is most commonly seen in one knee joint. The recommended treatment is total synovectomy, while radiotherapy can be used as adjuvant therapy for patients at risk for recurrence. The aim of our study was to show that the devastating effects of inactive diffuse PVNS may not be recognized for years and to evaluate the efficiency of aggressive total synovectomy on patients with PVNS during a follow-up period of 5 years. In the present study, 5 knees of four patients who had been previously followed due to gonarthrosis and diagnosed with PVNS during total knee arthroplasty (TKA) were followed and evaluated for a mean duration of 68 months. Mean age of the patients was 61.2 (52–66). All patients were women. One patient had right knee involvement, two had left knee involvement, and one had both knees involved. All patients had diffuse PVNS. Total synovectomy was performed in all patients in addition to TKA. During the follow-up, recurrence was not seen in any of the patients and prosthesis loosening was not detected. The aim of the present study was to evaluate the effectiveness of total synovectomy over the 5 years following the operation and to show that the devastating effects of inactive PVNS may sometimes be overlooked for years before being recognized during the treatment of the gonarthrosis that develops due to the disease. Although the disease is generally monoarticular, the study also presents a patient with bilateral PVNS.     

Plasma protein oxidation in aging rats after alpha-lipoic acid administration

In the present study, we investigated whether alpha-lipoic acid administration could have prooxidant or antioxidant effect on oxidative protein damage parameters such as protein carbonyl, nitrotyrosine, advanced oxidation protein products, and protein thiol, as well as oxidative stress parameters such as total thiol, nonprotein thiol, and lipid hydroperoxide in the plasma proteins of aged rats. Alpha-lipoic acid (100 mg/kg body wt/day) was administrated intraperitoneally to the Sprague-Dawley rats for 14 days. Protein carbonyl, nitrotyrosine, and advanced oxidation protein products levels were increased, protein thiol, nonprotein thiol, and total thiol levels were not changed in the plasma proteins of aged rats with alpha-lipoic acid administration. In aging rats with and without alpha-lipoic acid administration, plasma lipid hydroperoxide levels were significantly increased compared with those of the control group. The increased levels of protein oxidation markers such as protein carbonyl, nitrotyrosine and advanced oxidation protein products in the plasma proteins of alpha-lipoic acid-administrated aged rats compared with nonadministrated aged rats suggests that protein oxidation is increased in alpha-lipoic acid-administrated aged rats. We assume that an explanation for our findings regarding alpha-lipoic acid administration on protein oxidation markers in the plasma proteins of aged rats may be due to the prooxidant effects of alpha-lipoic acid.     

Emricasan to prevent new decompensation in patients with NASH-related decompensated cirrhosis

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