Comparison between different bone treatments on areal bone mineral density (aBMD) and bone microarchitectural texture as assessed by the trabecular bone score (TBS).

PURPOSE: The objective of the study was to assess longitudinal effects of different osteoporosis treatments on TBS and aBMD at lumbar spine. METHOD: We analyzed 390 patients (men: 72; women: 318; age>40 years; mean follow-up of 20 months and BMI<37 kg/m(2)). We stratified the cohort by treatments: Naive of treatment (Naive, n=67), Calcium and Vitamin D (CaVitD, n=87), Testosterone (Te, n=36), Alendronate (AL, n=88), Risedronate (Ri, n=39), Denosumab (Dmb, n=43) and Teriparatide (PTH, n=30). The follow-up changes from baseline were normalized at 24 months. RESULTS: After 24 months, Naive group TBS decreased by 3.1% (p<0.05) whereas a non-significant increase was observed for spine aBMD (Δ=+0.5%). Compared to the Naive group, significant improvement (p<0.05) was observed in both TBS and aBMD for Te, AL, Ri, Dmb and PTH groups and in the CaVitD group for TBS. At the end of the follow-up, significant improvement have been observed for aBMD in Te (+4.4%), AL (+4.1%), Ri (+4.8), D (+8.8%) and PTH (+8.8%) groups. Significant improvement was observed only in the AL (+1.4%), Dmb (+2.8%) and PTH (+3.6%) groups for TBS. CONCLUSION: As expected, TBS of Naive subjects decreased with age. As expected a TBS preservation has been observed under AL and Ri. Te and CaVitD effects on TBS were evaluated for the first time: a similar preservation effect has been observed. A significant TBS increase was observed under Denosumab and PTH. TBS could be a useful tool to monitor treatment effects.

Evaluating the efficacy of cinnamaldehyde and Echinacea purpurea plant extract in broilers against Eimeria acervulina.

Coccidiostats could be phased out as feed additives before 1 January 2013 for public health and food safety reasons, and, as a replacement, bioactive compounds found in plants are currently being investigated since they are more likely to be found acceptable by consumers. The aim of our study was to evaluate the effect of cinnamaldehyde (CIN) and Echinacea purpurea plant extract (EP) as additives by analyzing the performance traits, oocyst excretion and intestinal lesions following experimental infection with Eimeria acervulina. A total of 72 Ross male broilers were raised from 1 to 35 d and randomly assigned to four dietary treatments: control, without additives (C); 150 mg kg(-1) cinnamaldehyde (CIN); 1000 mg kg(-1)E. purpurea plant extract (EP); 150 mg kg(-1) cinnamaldehyde plus 1000 mg kg(-1)E. purpurea plant extract (CIN+EP). At 25 d, 12 chickens per treatment were orally infected with E. acervulina. Coccidia infestation led to lower performance but with no significant differences between the infected groups. Oocyst output reached its peak from 6 to 9 d post-infection in all treatments. At duodenal level, gross lesion scores were lower for cinnamaldehyde diets (P<0.05). A similar trend was observed in the microscopic lesion scores, with a non-significant reduction as a result of cinnamaldehyde addition (P>0.05). Scoring methods for macro- and microscopic lesions showed a positive linear relationship (G=+0.70). Further studies are necessary to assess the possible anticoccidian action of the cinnamaldehyde and its value as an alternative or adjunct in therapeutic or prophylactic strategies.

Immunocytochemical localization of copper,zinc superoxide dismutase in peroxisomes from watermelon (Citrullus vulgaris Schrad.) cotyledons.

In previous works using cell fractionation methods we demonstrated the presence of a Cu,Zn-containing superoxide dismutase in peroxisomes from watermelon cotyledons. In this work, this intracellular localization was evaluated by using western blot and EM immunocytochemical analysis with a polyclonal antibody against peroxisomal Cu,Zn-SOD II from watermelon cotyledons. In crude extracts from 6-day old cotyledons, analysis by western blot showed two cross-reactivity bands which belonged to the isozymes Cu,Zn-SOD I and Cu,Zn-SOD II. In peroxisomes purified by sucrose density-gradient centrifugation only one cross-reactivity band was found in the peroxisomal matrix which corresponded to the isozyme Cu,Zn-SOD II. When SOD activity was assayed in purified peroxisomes two isozymes were detected, Cu,Zn-SOD II in the matrix, and a Mn-SOD in the membrane fraction which was removed by sodium carbonate washing. EM immunocytochemistry of Cu,Zn-SOD on sections of 6-day old cotyledons, showed that gold label was mainly localized over plastids and also in peroxisomes and the cytosol, whereas mitochondria did not label for Cu,Zn-SOD.

Evidence for the presence of proteolytic activity in peroxisomes.

The presence of endo- and exoproteolytic activity in peroxisomes was detected in cell organelles purified from pea leaves. By PAGE using different exopeptidase substrates (L-aa-beta NA), one leucine aminopeptidase (AP) was found in peroxisomes. The peroxisomal AP was characterized as a serine protease and had a maximal activity at pH 7.5, a molecular mass of 56.8 kDa and a pI of 5.3. This enzyme was mainly present in the soluble fraction of peroxisomes. The occurrence of proteases in peroxisomes suggests that they might be involved in the protein turnover and processing of imported precursor polypeptides in peroxisomes.

Sodium fluoride prevents bone loss in primary biliary cirrhosis.

Low-bone-turnover osteoporosis is a common complication of primary biliary cirrhosis (PBC). Since sodium fluoride stimulates bone formation we assessed the effect of this drug on bone mass in a 2-year, prospective, double-blind trial including 22 women with PBC who were randomly assigned to receive sodium fluoride (50 mg/day) or placebo. All received calcium supplements and low doses of vitamin D. Bone mineral density of the lumbar spine was measured by dual-photon absorptiometry initially and every 6 months. Vertebral fractures were evaluated in thoracic and lumbar spine initially, and after 1 and 2 years. Seven patients in the fluoride group and eight in the placebo group completed the trial. In the fluoride group, bone mineral density did not change after 2 years (initial 1.05 +/- 0.07, final 1.07 +/- 0.06 g/cm2; p = n.s.). In the placebo group, however, bone mineral density decreased significantly (initial 1.00 +/- 0.07, final 0.93 +/- 0.06 g/cm2; p = 0.03). Moreover, in the fluoride group bone mineral density increased by 2.9 +/- 3.6%, and in the placebo group decreased by 6.6 +/- 2.6% (p = 0.04). None of the patients developed new vertebral or non-vertebral fractures. Treatment with sodium fluoride did not impair liver function or cholestasis in PBC. These results indicate that sodium fluoride prevents bone loss in PBC and therefore might be considered as a possible therapeutic agent for osteoporosis associated with this liver disease. Since a small number of patients completed the trial, further studies are required.

Peroxisome proliferation and oxidative stress mediated by activated oxygen species in plant peroxisomes.

The existence of a relationship between clofibrate-induced peroxisome proliferation and oxidative stress mediated by activated oxygen species was studied in intact peroxisomes purified from Pisum sativum L. plants. Incubation of leaves with 1 mM clofibrate produced a remarkable increase in the peroxisomal activity of acyl-CoA oxidase and, to a lesser extent, of xanthine oxidase, whereas there was a nearly complete loss of catalase activity and a decrease in Mn-superoxide dismutase. Ultrastructural studies of intact leaves showed that clofibrate induced a five- and twofold proliferation of the peroxisomal and mitochondrial populations, respectively, in comparison with those in control leaves. Prolonged incubation with clofibrate produced considerable alterations in the ultrastructure of cells. In peroxisomal membranes, the NADH-induced generation of O2- radicals, as well as the lipid peroxidation of membranes, increased as a result of treatment of plants with clofibrate. In intact peroxisomes treated with this hypolipidemic drug, the H2O2 concentration was higher than in peroxisomes from control plants. These results demonstrate that clofibrate stimulates the production of activated oxygen species (O2- and H2O2) inside peroxisomes, as well as the lipid peroxidation of peroxisomal membranes. This effect is concomitant with a decrease of catalase and Mn-SOD activities, the main peroxisomal enzymatic defenses against H2O2 and O2-, and indicates that in the toxicity of clofibrate, at the level of peroxisomes, an oxidative stress mechanism mediated by activated oxygen species is involved.

Treatment of steroid-induced osteopenia with calcitonin in corticosteroid-dependent asthma. A one-year follow-up study.

Sixty-two steroid-dependent asthmatics who had not received any form of treatment to prevent bone loss were studied during a 12-month period. Patients were randomly divided into two groups. Thirty-one patients were treated with 1 g of elemental calcium taken daily plus 100 IU of salmon calcitonin every other day, administered subcutaneously; the remaining 31 patients received only calcium supplementation. In the calcitonin group, 11 patients dropped out of the study because of severe side effects (seven patients), lack of compliance (three patients), and exacerbation of asthma (one patient). The 20 patients who completed the 12-month follow-up period were analyzed and compared with 20 sex-matched patients from the control group. At one year, bone mineral density (BMD) had increased in the calcitonin group by a mean of 4% (p less than or equal to 0.001), whereas in the control group BMD had decreased by 2.5% (p less than or equal to 0.05). Parameters of bone remodeling (alkaline phosphatase and osteocalcin) decreased significantly in the calcitonin-treated group but not in the control group. Our findings show that calcitonin 100 IU, given three times/wk, is an effective drug in the treatment of steroid-induced osteopenia. Side effects, however, are frequent and cause a high degree of dropout from therapy. These findings suggest that further studies should be carried out with lower doses of calcitonin or by other better tolerated forms of delivery such as in a nasal spray.

Percoll reversibly inhibits superoxide dismutase.

Incubation of pea leaf extracts (Pisum sativum L.) at 6 degrees C in isoosmotic media containing different Percoll concentrations significantly represses the total superoxide dismutase (SOD) activity in a concentration- and time-dependent manner. After 24 h incubation at 6 degrees C, 30-45% Percoll concentrations bring about an inhibition of Mn-SOD activity of more than 50%. Isozyme Cu,Zn-SOD II is affected to a lesser extent, with a maximum inhibition of 36% at high Percoll concentrations, whereas isozyme Cu,Zn-SOD I undergoes only slight variations. However, dilution of the samples followed by electrophoresis completely removes the Percoll inhibitory action. Results suggest that superoxide dismutases could be adsorbed onto the Percoll surface through electrostatic interactions.