Search

In an ethnically homogeneous population of women living in Tuscany, Italy, the relationships between age, body weight, bone mineral density and the vitamin D receptor (VDR) gene polymorphism were studied, with the objective of recognizing patients at risk for osteoporosis.  In 275 women bone mineral density was measured by Dual Energy X-rays Absorptiometry (DEXA).  In 50 of them the individual genetic pattern for VDR was evaluated by DNA extraction followed by PCR amplification of the VDR gene, and digestion with the restriction enzyme BsmI. Age and bone mineral density were inversely related (R2 = 0.298).  Body weight was associated with bone mineral density (R2 = 0.059), but not with age.  In osteoporotic women, mean (± SD) body weight was 59.9 ± 6.5 Kg, lower than that recorded in non osteoporotic women (64.2 ± 9.4 Kg), even though not significantly different (p = 0.18).  No association was found between VDR gene polymorphism, bone density or body weight.  The performance of anthropometric and genetic components appear to be poor, and, at least for the time being, bone mineral density measurement by means of MOC-DEXA represents the optimal method to detect women at risk for postmenopausal osteoporosis. [ABSTRACT FROM AUTHOR]

Rab GDP-dissociation inhibitors (GDI) are evolutionarily conserved proteins that play an essential role in the recycling of Rab GTPases required for vesicular transport through the secretory pathway. We have found mutations in the GDI1 gene (which encodes αGDI) in two families affected with X-linked non-specific mental retardation. One of the mutations caused a non-conservative substitution (L92P) which reduced binding and recycling of RAB3A, the second was a null mutation. Our results show that both functional and developmental alterations in the neuron may account for the severe impairment of learning abilities as a consequence of mutations in GDI1, emphasizing its critical role in development of human intellectual and learning abilities. [ABSTRACT FROM AUTHOR]

The article discusses a study conducted to evaluate in the human cerebellum the presence of an intrinsic serotonergic neuronal subpopulation. The immunoreactions were revealed by streptavidin-biotin-peroxidase and 3,3-diaminobenzidine tetrahydrochloride technique; for positive controls were used duodenum and brainstem fragments of rat subjected to the same experimental protocol.

C Cadmium is a toxic heavy metal known as environmental pollutant world-wide distributed. Its toxicity, predominantly due to its long half-life of 15-20 years, affects many organs and tissues including kidney, lung, liver, heart, bone, muscles and the central nervous system (CNS) as well [1]. Our research group previously demonstrated that cadmium is able to alter the blood-brain barrier by modifying the tight junction sealing morphology and to induce oxidative stress, thus leading to neuronal degeneration mainly by oxidative stress [2]. Among the cellular population of the CNS, glial cells own a pivotal role in maintaining the physiological brain homeostasis and their alteration has been demonstrated to indirectly affect neurons [3]. The aim of the present research is to evaluate cadmium toxicity in two glial cell lines (astrocyte DITNC1 and microglia BV-2), at different time and increasing concentration of metal exposure. Cadmium increasing concentrations (0.1-100 µM) were administered in starvation medium for 8, 24 and 48 hours. Cell viability assay, western blotting analysis, and immunofluorescent staining were employed in order to evaluate the molecular and morphological alterations induced by the heavy metal. Our results clearly demonstrated that Cd is able to affect the cell viability even at the lowest concentrations and in a time-dependent manner, by altering the oxidative balance (ROS production, cytochrome C expression) and inducing an endoplasmic reticulum stress (GRP78 upregulation). Moreover, Cd is able to induce glial morphological alterations as well as an overexpression of activation glial markers, such as GFAP, S100ß, CD86, and Iba1. In conclusion, the data obtained clearly demonstrated that Cd toxicity occurs even at low doses and for very short exposure times Further experiments should be performed in order to evaluate the mechanisms underlying the Cd-dependent activation of glial cells and how this, in turn, induces a neuronal homeostasis alteration. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is a highly toxic environmental pollutant released from the smelting and refining of metals and cigarette smoking. Oral exposure to cadmium may result in adverse effects on a number of tissues, including the central nervous system (CNS). In fact, its toxicity has been related to neurological disorders, as well as neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. Under normal conditions, Cd barely reaches the brain in adults because of the presence of the blood–brain barrier (BBB); however, it has been demonstrated that Cd-dependent BBB alteration contributes to pathogenesis of neurodegeneration. However, the mechanism underlying Cd-dependent BBB alteration remain obscure. Here, we investigated the signaling pathway of Cd-induced tight junction (TJ), F-actin, and vimentin protein disassembly in a rat brain endothelial cell line (RBE4). RBE4 cells treated with 10 μM cadmium chloride (CdCl2) showed a dose- and time-dependent significant increase in reactive oxygen species (ROS) production. This phenomenon was coincident with the alteration of the TJ zonula occludens-1 (ZO-1), F-actin, and vimentin proteins. The Cd-dependent ROS increase elicited the upregulation of GRP78 expression levels, a chaperone involved in endoplasmic reticulum (ER) stress that induces caspase-3 activation. Further signal profiling by the pannexin-1 (PANX1) specific inhibitor 10Panx revealed a PANX1-independent increase in ATP spillage in Cd-treated endothelial cells. Our results point out that a ROS-dependent ER stress-mediated signaling pathway involving caspase-3 activation and ATP release is behind the BBB morphological alterations induced by Cd. [ABSTRACT FROM AUTHOR]

Sonic hedgehog (Shh) signaling is a key pathway within the central nervous system (CNS), during both development and adulthood, and its activation via the 7-transmembrane protein Smoothened (Smo) may promote neuroprotection and restoration during neurodegenerative disorders. Shh signaling may also be activated by selected glucocorticoids such as clobetasol, fluocinonide and fluticasone, which therefore act as Smo agonists and hold potential utility for regenerative medicine. However, despite its potential role in neurodegenerative diseases, the impact of Smo-modulation induced by these glucocorticoids on adult neural stem cells (NSCs) and the underlying signaling mechanisms are not yet fully elucidated. The aim of the present study was to evaluate the effects of Smo agonists (i.e., purmorphamine) and antagonists (i.e., cyclopamine) as well as of glucocorticoids (i.e., clobetasol, fluocinonide and fluticasone) on NSCs in terms of proliferation and clonal expansion. Purmorphamine treatment significantly increased NSC proliferation and clonal expansion via GLI-Kruppel family member 1 (Gli1) nuclear translocation and such effects were prevented by cyclopamine co-treatment. Clobetasol treatment exhibited an equivalent pharmacological effect. Moreover, cellular thermal shift assay suggested that clobetasol induces the canonical Smo-dependent activation of Shh signaling, as confirmed by Gli1 nuclear translocation and also by cyclopamine co-treatment, which abolished these effects. Finally, fluocinonide and fluticasone as well as control glucocorticoids (i.e., prednisone, corticosterone and dexamethasone) showed no significant effects on NSCs proliferation and clonal expansion. In conclusion, our data suggest that Shh may represent a druggable target system to drive neuroprotection and promote restorative therapies. [ABSTRACT FROM AUTHOR]

Interaction between intestinal epithelial cells (IECs) and the underlying immune systems is critical for maintaining intestinal immune homeostasis and mounting appropriate immune responses. We have previously showed that the T helper type 1 (TH1) cytokine IL-12 plays a key role in the delicate immunological balance in the gut and the lack of appropriate levels of IL-12 had important consequences for health and disease, particularly with regard to food allergy. Here we sought to understand the role of IL-12 in the regulation of lympho-epithelial cross talk and how this interaction affects immune responses locally and systemically. Using a combination of microscopy and flow cytometry techniques we observed that freshly isolated IECs expressed an incomplete, yet functional IL-12 receptor (IL-12R) formed solely by the IL-12Rb2 chain that albeit the lack of the complementary IL-12b1 chain responded to ex-vivo challenge with IL-12. Furthermore, the expression of IL-12Rb2 on IECs is strategically located at the interface between epithelial and immune cells of the lamina propria (lp) and using in vitro co-culture models and primary intestinal organoids we showed that immune-derived signals were required for the expression of IL-12Rb2 on IECs. The in vivo biological relevance of the IEC-associated IL-12Rb2 was assessed in vivo in a mouse model of food allergy characterized by allergy-associated diminished intestinal levels of IL-12 and in chimeric mice that lack the IL-12Rb2 chain on IECs. These experimental models enabled us to show that the anti-allergic properties of orally delivered recombinant Lactococcus lactis secreting bioactive IL-12 (rLc- IL12) were reduced in mice lacking the IL-12b2 chain on IECs. Finally, we observed that the oral delivery of IL-12 was accompanied by the down-regulation of the production of the IECderived pro-allergic cytokine thymic stromal lymphopoietin (TSLP). However, further analysis of intestinal levels of TSLP in IL-12Rb2-/- mice suggested that this event was not directly linked to the IEC-associated IL-12Rb2 chain. We interpreted these data as showing that IECassociated IL12Rb2 is a component of the cytokine network operating at the interface between the intestinal epithelium and immune system that plays a role in immune regulation. [ABSTRACT FROM AUTHOR]

It is known that joint mobility (JM), flexibility, muscle strength and posture can determine the quality of movement and could also adversely affect the body development of young subjects practicing sport. The aim of this study was to investigate how the practice of different sports affects these parameters. We enrolled 109 young subjects practicing different sports: soccer (M/F:22/10), mean age 12,6±1,4 yrs, BMI:18,3±2,3 kg/m2; volleyball (M/F:13/14), mean age 12,0±1,3 yrs, BMI 17,4±2,2 kg/m2; basketball (M/F:20/2), mean age 11,1±0,8 yrs, BMI 20,9±4,4 kg/m2; gymnastics (M/F:0/11), mean age 14,5±1,1 yrs, BMI 18,9±2,1 kg/m2 and dance (M/F:0/17), mean age 11,7±3,1 yrs, BMI 18,3±2,8 kg/m2. In these subjects we evaluated ankle JM (inclinometer), trunk flexibility (sit & reach test), muscle strength (standing long jump, vertical jump and Jamar hand grip), posture (images on the sagittal plane) and lifestyle (IPAQ-C, IPAQ-A). The individual sporting history was investigated by a specific questionnaire. The tests performed showed a significantly higher ankle JM in young dancers (155,8±10,3°) compared to all other groups excluding volleyball players (p<0,001). In particular, the subjects practicing soccer showed a significant reduction of the ankle JM (125,2±22,3°) compared to all the other groups of subjects investigated (p<0,01). Gymnasts showed a greater flexibility of the trunk than that measured in all other groups (18,3±3,5 cm; p<0,001), while basketball players showed lower trunk flexibility (-7.7 ± 7.0 cm). In the muscle-strength tests performed the dancers showed the following results (hand grip: 18.7 ± 6.6 kg, long-jump standing 119.8 ± 29.2 cm) that are significantly reduced compared to the gymnasts (hand grip: 26.0 ± 4, 2 kg: p <0.005, long jump standing 163.8 ± 12.7 cm: p <0.001) and to volleyball players (standing long jump 152.5 ± 27.9 cm: p <0.001). The practice of specific sports can significantly modify the ankle JM and the flexibility of the trunk as well as affect muscle strength even in young subjects. It is not entirely clear whether these effects may induce negative consequences on health and development of the anatomical structures involved; therefore, further studies are needed to verify the conclusions and the possible usefulness of APA programmes in the prevention or recovery of such significant effects. [ABSTRACT FROM AUTHOR]

Exercise is generally recommended for elderly subjects although the effects of the different programmes used in the adapted physical activity (APA) centers are not always properly verified. The aim of this study was to evaluate the response of elderly subjects attending classes of APA to a moderate aerobic physical activity. In 15 subjects (2/13 M/F), mean age 67,8±13,8 yrs, body mass index 23,7±3,5 Kg/m2 attending an APA class we evaluated ankle joint mobility (inclinometer), hand strength (Jamar hand grip), walking speed and step length (10 and 20 meters), aerobic capacity and endurance (6 Minute Walk Test- 6MWT), lower extremity function (SPPB: short physical performance battery), posture (sagittal images), and peripheral microcirculation (Laser doppler flowmetry). The subjects were evaluated in the following 3 stages: at the end of a 8-months APA period, after 4 months of inactivity and 4 months after recommencing of a new period of the same programme. All the subjects included in the study performed a training programme of APA twice a week for 1 hour each. The training programme consisted in 10 minutes of organic activation, 30 minutes of moderate physical activity at 45-65% of VO2 peak and toning, 20 minutes of exercises on the floor: breathing and stretching. After 4 months of inactivity, the subjects investigated showed a significant and widespread reduction of the gait parameters investigated: 6MWT (446.5 ± 91.4 vs 429 ± 89.4 mt, p <0.01); 10 meters (step length: 0.67 ± 0.09 vs 0.63 ± 0.08 mt - time: 7.6 ± 1.7 vs 8.5 ± 1.7''); 20 meters (step length: 0.71 ± 0.08 vs 0.67 ± 0.06 mt - time: 14.0 ± 2.1 vs 15.4 ± 2.0''). After 4 months of training there was a significant, even if partial recovery of the parameters investigated in comparison to the results achieved after the inactivity period,: 6MWT (438.05 ± 92.3 mt, p <0.01); 10 meters (step length: 0.65 ± 0.07 mt - time: 8.0 ± 1.7'', p <0.01); 20 meters (step length: 0.68 ± 0.07 mt, p = 0.15 - time: 14.6 ± 2.1'', p <0.01). The 4 meter walking speed (SPPB) evaluated in the subjects investigated was correlated with 10 mt (r = 0.69, p <0.01) and 20 mt (r = 0.6, p <0, 05). The length of the step measured in the 10 and 20 meters tests was correlated with the ankle mobility (10 mt: r = 0.7, p <0.01; 20 mt: r = 0.62, p <0.5). The results of this study show that in a population of elderly subjects a programme of APA, as scheduled and performed in this study, significantly improves the gait parameters. However, a short interruption of the APA training significantly reduced the walking speed and step length. This study, in addition to underlining the importance of the APA programmes, also shows that it is necessary to reduce or avoid interruptions of these physical activities. [ABSTRACT FROM AUTHOR]

Degenerative diseases of the central nervous system are significant causes of mortality among aging population in industrialized countries, as well as expensive for the national health systems and caregivers [1]. Nowadays, pharmacological therapies are still not decisive because they are hindered by the blood brain barrier (BBB) [2]. Although in the last decades many researchers have conducted in vivo experiments to better identify a therapeutic approach of ultrasound (US) in neurodegenerative diseases, little is known about their role in increasing the BBB permeability [3,4]. The present study aims to identify, the effect of focused US on a rat brain endothelial cell line (RBE4). After ultrasound stimulation (10-20-30 min.), MTT and western blotting assay were conducted to demonstrate the non-toxic effects of US. Furthermore, immunostaining of stimulated cells was performed to detect changes in cytoskeletal F-actin fibers and Zonula occludens-1 (ZO1) tight juntion. BBB opening was evaluated by measuring the extent of the intercellular space, in Papanicolaou stained cells. The results evidenced an ultrasonic-dependent mechanical action on F-actin fibers that altered their distribution within the cells showing the formation of numerous stress fibers. F-actin alterations were accompanied by an alteration of the ZO-1 distribution, exhibiting a "zipper-like" staining pattern and holes that became visible between cells. Papanicolaou staining confirm the opening of the BBB evidencing many wider areas of intercellular space. All these structural changes on RBE4 cell line occurred without significant alterations in metabolic activity as well as in absence of apoptotic or endoplasmic reticulum stress markers. In conclusion, these results confirm and highlight the potential role of ultrasound in the permeabilization of the BBB, thus suggesting new ways for drugs administration. [ABSTRACT FROM AUTHOR]

Quality of life into later life is influenced by multiple factors. The physical abilityto perform common everyday activities represents a key factor to maintain a healthy and independent lifestyle. Moreover, aging is a process characterized by physiological alterations resulting in a progressive decline in biological functions, decreased resistance to stress, and increased susceptibility to diseases. Especially in elderly people, alterations such as imbalance between pro and antioxidant activity and/or hormonal dysregulation negatively affect the physical capacity, the emotionalstatus and the overall general health and quality of life [1]. On the other hand, regular physical activity is considered an effective strategy for older adults to prevent and reduce the risk of developing those negative conditions arising from aging. A 24-week regular physical activity program (twice weekly, 1 hour per session) focused on functional fitness exercises was performed by 20 older adults (aged 55 years or more). A set of anthropometric (height, weight, BMI and body fat percentage) and physical measurements (grip strength, chair sit to stand, sit and reach and back scratch) assessing the functional fitness performance [2] were evaluated. Moreover, biochemical markers (d-ROMs and BAP tests as assessment of oxidative stress and antioxidant potential; salivary cortisol levels) were measured before and after the intervention program. The results confirm the benefits of a regular physical activity in older adults resulting in improved physical strength and flexibility in the functional fitness parameters, and in regulating pro and antioxidant activity and cortisol levels. [ABSTRACT FROM AUTHOR]

During infection intestinal CX3CR1+ cells can either extend transepithelial cellular processes to sample luminal bacteria or, very early after infection migrate into the intestinal lumen to capture bacteria. However, up to date, the biological relevance of the intraluminal migration of CX3CR1+ cells remained to be determined. We addressed this by using a combination of mouse strains differing in their ability to carry out CX3CR1-mediated sampling and intraluminal migration. We observed that, the number of S. Typhimurium traversing the epithelium did not differ between sampling-competent/migration-competent C57BL/6 and sampling deficient/migration- competent Balb/c mice. By contrast, in sampling-deficient/migration deficient CX3CR1-/- mice the numbers of S. Typhimurium penetrating the epithelium were significantly higher. However, in these mice the number of invading S. Typhimurium was significantly reduced after the adoptive transfer of CX3CR1+ cells directly into the intestinal lumen, consistent with intraluminal CX3CR1+ cells preventing S. Typhimurium from infecting the host. This interpretation was also supported by a higher bacterial faecal load in CX3CR1+/gfp compared to CX3CR1gfp/gfp mice following oral infection. Furthermore, by using real time in vivo imaging we observed that CX3CR1+ cells migrated into the lumen moving through paracellular channels within the epithelium. Also, we reported that the absence of CX3CR1 mediated sampling did not affect antibody responses to a non-invasive S. Typhimurium strain that specifically targeted the CX3CR1-mediated entry route. These data showed that the rapidly deployed CX3CR1+ cell-based mechanism of immune-exclusion is a defence mechanism against pathogens that complements the mucous and secretory (s)IgA antibodymediated system in the protection of intestinal mucosal surface. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is a worldwide occupational and environmental pollutant [1]. Cd toxicity is widely known and studied in many tissues and organs such as testis, kidney, liver, lung and brain [2,3]. On the other hand, zinc (Zn) is a trace element known as coenzyme for many proteins such as methallothionein [4]. To date, very little is known about the role of Cd and Zn in BBB permeability. To study their effects in BBB permeability in vitro, the RBE4 cell line was used and different concentrations of CdCl2 and ZnCl2 were tested. Metabolic activity (MTT assay) was performed to test the protective and preventive role of ZnCl2 on CdCl2 toxicity. Western blotting analysis was used to better investigate the molecular pathway involved in Cd-induced BBB permeability evaluating GRP78 (ER stress marker) and caspase-3 protein expression levels. Furthermore, ZO-1 and F-actin immunofluorescent staining was performed to better understand the morphological alterations and BBB permeability achieved by Cd treatment. Our preliminary data highlight the role of Cd in evoking BBB permeability by F-actin and ZO1 dislocations, triggering the caspase-3 molecular pathway activation induced by GRP78-ER stress increase. Moreover, the data clearly show how Zn is able to counteract the metabolic impairment induced by Cd treatment. Taken together these data point out the possible role of Zn in counteracting the Cd-induced BBB impairment. [ABSTRACT FROM AUTHOR]

Due to its high permeability to blood-brain barrier (BBB), cadmium (Cd) has been regarded as a possible etiological factor for human neurodegenerative diseases, such as Parkinson's disease, Alzheimer's disease, and Huntington's disease [1]. However, the mechanism underlying cadmium-induced BBB permeability remains unclear. In this study, we investigated the effect of Cd in RBE4 cells (rat brain endothelial cells) and delineated the signaling pathway that, triggering endoplasmic reticulum (ER) stress and caspase 3 activation, leads to cytoskeleton disorganization and tight junctions disassembly. Our results demonstrate a possible downstream pathway mediated through the Cd-dependent ER stress, assessed by the cytoplasmic expression of an ER protein GRP78, and the consequent caspase-3 activation that results in an extracellular ATP increase, which in turns induces a dislocation, evaluated by immunofluorescent staining, of Zonula Occludens-1 (ZO-1), a tight junction protein, and F-actin. These findings, whereby Cd-induced permeabilization of BBB through a ER stressdependent pathway on endothelial cells represent a possible novel mechanism of action for Cd that could explain, at least in part, the Cd-related central effects. [ABSTRACT FROM AUTHOR]

19, 134?139 (1998). [ABSTRACT FROM AUTHOR]