Giovanni Tringali

Increased sclerostin serum levels associated with bone formation and resorption markers in patients with immobilization-induced bone loss.

CONTEXT Sclerostin, a Wnt signaling antagonist on the osteoblasts produced by osteocytes, is regulated by mechanical strain and is implicated in the pathogenesis of disuse bone loss. There are no data on sclerostin in humans. OBJECTIVE The aim of the study was to evaluate sclerostin in patients immobilized after stroke, compared with control subjects, and to analyze its relationship with markers of bone formation and resorption. DESIGN This was a cross-sectional study. SETTING AND PATIENTS We studied 40 postmenopausal women immobilized after a single episode of stroke 6 months or longer after onset, and 40 postmenopausal women from the general community. Bone status was assessed by quantitative ultrasound measurements at the calcaneus. Bone alkaline phosphatase (b-AP), carboxy-terminal telopeptide of type I collagen (CrossLaps), and sclerostin were evaluated by ELISA. We also used ELISA to measure serum levels of Dickkopf-1, another soluble inhibitor of Wnt/beta-catenin signaling, highly expressed by osteocytes. RESULTS Immobilized patients had higher sclerostin serum levels (median 0.975 ng/ml; 25th to 75th percentiles 0.662-1.490) than controls (median 0.300 ng/ml; 25th to 75th percentiles 0.165-0.400: P < 0.0001) and an increased bone turnover with a more significant rise in bone resorption (CrossLaps) than formation (b-AP) markers. Sclerostin correlated negatively with b-AP (r = -0.911; P < 0.0001) and positively with CrossLaps (r = 0.391; P = 0.012). Dickkopf-1 did not significantly differ between the groups. Patients also had quantitative ultrasound measurements index lower than controls (P < 0.001). CONCLUSIONS This study shows for the first time that long-term immobilized patients present hypersclerostinemia associated with reduced bone formation, and suggests that sclerostin could be a link between mechanical unloading and disuse osteoporosis in humans.

Sclerostin levels associated with inhibition of the Wnt/β-catenin signaling and reduced bone turnover in type 2 diabetes mellitus.

CONTEXT Patients with type 2 diabetes (T2DM) have low bone turnover, poor bone quality, and circulating levels of sclerostin significantly higher than non-T2DM controls. There are no data on the possible association of sclerostin with β-catenin, a key component of the Wnt/β-catenin canonical signaling. OBJECTIVES The aim of the study was to evaluate the circulating β-catenin levels in T2DM patients and to analyze their relationship with sclerostin and bone turnover markers. DESIGN This was a cross-sectional study. SETTING AND PATIENTS The study was conducted at a clinical research center. Forty T2DM postmenopausal women were studied and compared with 40 healthy controls. Bone status was assessed by dual-energy x-ray absorptiometry measurements (bone mineral density) and by measuring bone alkaline phosphatase and carboxy-terminal telopeptide of type 1 collagen. Sclerostin and β-catenin were evaluated by an immunoenzymetric assay. RESULTS Consistent with previous reports in T2DM subjects, we found sclerostin levels higher and bone turnover markers lower than controls. In our cohort of T2DM patients, β-catenin levels are significantly lower than in controls (median 1.22 pg/ml, 25th to 75th percentiles 0.50-2.80; and median 4.25 pg/ml, 25th to 75th percentiles 2.20-7.62, respectively; P=0.0002). β-Catenin correlated negatively with sclerostin (P<0.0001) and positively with bone alkaline phosphatase (P=0.0030) only in T2DM patients and negatively with age in both groups. Eight of the 40 T2DM patients had vertebral fractures. CONCLUSIONS These results show for the first time that T2DM patients have serum concentrations of β-catenin lower than controls. The negative association of β-catenin with sclerostin suggests a biological effect of increased sclerostin on the Wnt signaling, which appears impaired in T2DM.

Effect of ischemia-reperfusion on renal expression and activity of N(G)-N(G)-dimethylarginine dimethylaminohydrolases.

Background:Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase. It is degraded by the enzyme dimethylarginine dimethylaminohydrolase (DDAH). Methods:Rats (n = 50) underwent to 45 min of renal ischemia followed by 30 min, 1 h, and 3 h of reperfusion. Expression of endothelial nitric oxide synthase, inducible nitric oxide synthase, DDAH-1, DDAH-2, renal DDAH activity, plasma NO2−/NO3−, and ADMA levels were evaluated. Results:Inducible nitric oxide synthase expression increased, as confirmed by both plasma (11.89 ± 1.02, 15.56 ± 0.93, 11.82 ± 0.86, 35.05 ± 1.28, and 43.89 ± 1.63 nmol/ml in the control, ischemic, 30-min, 1-h, and 3-h groups, respectively) and renal (4.81 ± 0.4, 4.85 ± 1, 9.42 ± 0.7, 15.42 ± 0.85, and 22.03 ± 1.11 nmol/mg protein) formations of NO2−/NO3−. DDAH-1 expression decreased after reperfusion, whereas DDAH-2 increased after 30 min, returning to basal levels after 3 h. Total DDAH activity was reduced during all times of reperfusion. Both plasma (0.41 ± 0.03, 0.43 ± 0.05, 0.62 ± 0.02, 0.71 ± 0.02, and 0.41 ± 0.01 nmol/ml in the control, ischemic, 30-min, 1-h, and 3-h groups, respectively) and renal (1.51 ± 0.01, 1.5 ± 0.01, 1.53 ± 0.01, 2.52 ± 0.04, and 4.48 ± 0.03 nmol/mg protein in the control, ischemic, 30-min, 1-h, and 3-h groups, respectively) concentrations of ADMA increased. Conclusions:Results suggest that ischemia–reperfusion injury leads to reduced DDAH activity and modification of different DDAH isoform expression, thus leading to increased ADMA levels, which may lead to increased cardiovascular risk.

The association between carotid or femoral atherosclerosis and low bone mass in postmenopausal women referred for osteoporosis screening. Does osteoprotegerin play a role

Atherosclerosis and osteoporosis appear to be epidemiologically correlated. Most (but not all) animal and clinical studies suggest that osteoprotegerin (OPG) may represent a possible molecular link between bone loss and vascular calcification. The aim of this study was to investigate the association of OPG with bone mineral density (BMD) and vascular plaques, in order to contribute to a better understanding of the link between atherosclerosis and osteoporosis. The study population consisted of 100 consecutive postmenopausal women referred for routine osteoporosis screening. BMD was evaluated by dual-energy X-ray absorptiometry. Presence of carotid or femoral plaques was examined by ultrasonography. OPG was measured by enzyme immunoassay. Seventy-two subjects had low bone mass and were categorized as osteopenic (32) or osteoporotic (40). Fifty-two subjects had one or more atherosclerotic plaques at carotid or femoral level. Both lumbar spine and femoral BMD were associated with the number of plaques (r=-0.5370; p<0.0001, and r=-0.4423; p=0.0012, respectively), however only spine BMD remained significantly associated with the number of plaques after adjustment. OPG serum values showed a significant association with age (r(2)=0.057; p=0.042). The association between OPG and the number of plaques was significant only in patients with concomitant involvement of carotid and femoral districts (r(2)=0.758; p<0.0001).

Genetic and environmental factors in human osteoporosis from Sub-Saharan to Mediterranean areas

The aim of this study was to determine the prevalence of known gene polymorphisms associated with osteoporosis in postmenopausal normal women from Burkina Faso and Sicily, compared to postmenopausal Sicilian women with osteoporosis, and to establish the weight of environmental factors in the mechanism of osteoporosis. Bone mass density (BMD) was measured by phalangeal quantitative ultrasound (QUS) in Burkinabe woman and by the dual X-ray absorptiometry at the femoral neck in Sicilian women. The polymorphisms of the vitamin D receptor (VDR) gene, estrogen receptor (ESR) gene, calcitonin receptor (CTR) gene and COL1A1 collagen gene were characterized by PCR. The social characteristics of studied women were evaluated by a specific questionnaire. The observed percentages of single specific polymorphisms did not differ from that expected with exception of VDR B allele and ESR X and P allele in Burkinabe and Sicilian women, respectively. Association analyses and multivariate two-step regression model of social and molecular parameters, demonstrated that in comparison to the VDR, ESR, CTR polymorphisms, physical activities and healthy diet, associated with outdoor work are the best favourable prognostic factors for osteoporosis. A diet rich in calcium, other minerals and vitamin D in association with physical activity represents the most effective way to maintain not only a healthy bone structure but also an acceptable BMD. This is particularly true for Sub-Saharan women.

Protective effects of a standardised red orange extract on air pollution-induced oxidative damage in traffic police officers.

Several pathological conditions have all been associated with a higher release of atmospheric pollutants. There is growing evidence that oxidative stress may represent one of the agents involved in the initiation and/or progression of many of these pathologies. The aim of the present study was to evaluate the effects of short-term dietary supplementation with a standardised red orange extract (ROC) on a group of traffic police officers exposed to traffic exhaust pollution and cigarette smoking, by measuring some noninvasive biomarkers of oxidative stress. At the beginning of the study, all the groups showed similar serum lipid hydroperoxide levels, but traffic officers showed lower serum concentrations of thiol (SH) groups; furthermore, the frequency of spontaneous sister chromatide exchanges (SCEs) in peripheral lymphocytes was increased by smoking (but not by pollution exposure alone) at a higher degree in subjects exposed to traffic pollution. After 1 month of ROC administration, serum lipid hydroperoxide levels decreased only in all non-smoking subjects; furthermore, SH group levels measured in traffic officers appeared restored to normal values observed in the respective controls. Finally, the increase in SCE frequency induced by smoking was reduced by treatment with ROC especially in traffic officers. Our study suggests that ROC supplementation could be useful to minimise the detrimental effects caused by exposure to air pollution and smoking.

Reduction of volumetric bone mineral density in postmenopausal women with hepatitis C virus-correlated chronic liver disease: A peripheral quantitative computed tomography (pQCT) study

BACKGROUND The prevalence of osteoporosis in chronic liver disease (CLD) varies considerably among the studies, depending on patient selection and diagnostic criteria. We aimed to measure bone turnover markers and volumetric bone mineral density (BMD) in a group of postmenopausal women with CLD using both dual energy X-ray absorptiometry (DXA) and peripheral quantitative computed tomography (pQCT), in comparison with age-matched healthy subjects. METHODS Thirty-five postmenopausal patients with HCV-correlated CLD and 35 healthy postmenopausal women, as controls, underwent a DXA scan at lumbar and femoral level and a pQCT measurement of the nondominant forearm. Serum concentrations of biochemical markers relevant to bone turnover were also measured. RESULTS Patients showed no differences in DXA values either at lumbar or femoral level compared to controls. On the contrary, pQCT geometrical (cortical cross-sectional area) and volumetric (total and trabecular volumetric BMD) parameters were significantly reduced in the CLD women. Also the Strength-Strain Index (SSI), an estimate of diaphyseal bone resistance to bending and torsion, was significantly lower in patients than in controls. Patients with CLD presented an unbalanced bone turnover, with increased bone resorption markers. CONCLUSIONS The bone geometrical and volumetric parameters measured in our CLD postmenopausal women, by pQCT, show a reduced bone mineral quality and stiffness. Conversely, DXA-measurements seem unable to appreciate the bone alterations in these patients. This would encourage further studies to validate pQCT analysis as a diagnostic tool for a correct estimate of bone involvement in CLD.

Basophil biomarkers as useful predictors for sublingual immunotherapy in allergic rhinitis

ABSTRACT Prevalence of allergic diseases is increasing worldwide. Allergen‐specific immunotherapy (ASIT) is potentially the only curative treatment for allergy, but there is a lack of reliable methods to monitor the immune responses to ASIT and to predict clinical efficacy. Recently, the definition of allergen sensitivity threshold (CD‐Sens) by Basophil Activation Tests has been suggested as potential method in this context. The aim of this study was to compare trends of CD‐Sens, measured by the markers CD63 and CD203c, and clinical symptoms in subjects with allergic rhinitis receiving Sublingual Immunotherapy (SLIT). 26 rhinitis patients allergic to Parietaria were selected and matched into two groups; a SLIT treated group (SG) and a reference group (RG) treated by traditional anti‐allergic medications. Visual Analogue Scale (VAS) score for the four cardinal symptoms of rhinitis and peripheral blood was collected before the first dose of SLIT (T0) and after 12months (T12) to define the severity of the symptoms and the sensitivity of basophils to Parietaria. The comparison between T0 and T12 in SG patients showed a significant decrease of symptom severity (VAS score) and an increased tolerability of basophils to Parietaria (CD‐Sens) both by CD63 and CD203c. But, only CD203c seems to be correlated with the clinical symptoms. These data corroborate the hypothesis that SLIT could change the immunological course of allergic sensitization already in the first year, and that an immunological parameter as CD‐Sens measured by CD63 and CD203c expression on stimulated basophils could be useful to monitor the changes in the immune system. HIGHLIGHTSThere is a lack of methods to predict the efficacy of allergen immunotherapy.The basophils threshold sensitivity by CD63/CD203c could be useful to this purpose.The marker CD203c seems to better related to clinical outcomes in allergic rhinitis.Allergen immunotherapy reduces the basophils sensitivity already in the first year.

Primary afferent plasticity following deafferentation of the trigeminal brainstem nuclei in the adult rat

Alpha-tyrosinated tubulin is a cytoskeletal protein that is involved in axonal growth and is considered a marker of neuronal plasticity in adult mammals. In adult rats, unilateral ablation of the left facial sensorimotor cortical areas induces degeneration of corticotrigeminal projections and marked denervation of the contralateral sensory trigeminal nuclei. Western blotting and real-time-PCR of homogenates of the contralateral trigeminal ganglion (TG) revealed consistent overexpression of growth proteins 15 days after left decortication in comparison with the ipsilateral side. Immunohistochemical analyses indicated marked overexpression of alpha-tyrosinated tubulin in the cells of the ganglion on the right side. Cytoskeletal changes were primarily observed in the small ganglionic neurons. Application of HRP-CT, WGA-HRP, and HRP to infraorbital nerves on both sides 15 days after left decortication showed a significant degree of terminal sprouting and neosynaptogenesis from right primary afferents at the level of the right caudalis and interpolaris trigeminal subnuclei. These observations suggest that the adaptive response of TG neurons to central deafferentation, leading to overcrowding and rearrangement of the trigeminal primary afferent terminals on V spinal subnuclei neurons, could represent the anatomical basis for distortion of facial modalities, perceived as allodynia and hyperalgesia, despite nerve integrity.