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The countryside hosts an increasing number of alternative food networks: rural tourists can play an important role in acting as both consumer and “cultural broker” between these networks. This paper provides a theoretical framework for niche marketing food specialties in rural tourism by combining two different consumer behavioural theories, the “experience economy” and the “intimacy” model, representing a reorientation from classical marketing thinking. It explores the meaning of local food, including the pursuit of reconnection with nature, resilience to globalisation, the role of local food in reinforcing personal identity, the search for freshness, taste and authenticity, support for local producers, and environmental concerns. It considers the challenges for rural entrepreneurs and policy makers in marketing food specialties and rural regions to the post-modern consumer. Using examples derived mostly from secondary literature it identifies seven dimensions that elevate food products to an appealing c...

Glial cells coordinate the differentiation, metabolism, and excitability of neurons; they modulate synaptic transmission and integrate signals emanating from neurons and other glial cells. Several evidences underlying the relation between these pathways and the regulatory mechanisms of ion concentration, supporting the role of Aquaporins (AQPs) in these processes. The goal of this review is to summarize the localization of different isoforms of AQPs in relation to glial cells both in central and peripheral nervous system, underlying AQP involvement in physiological and in pathophysiological conditions such as brain edema, glioma and epilepsy.

Glass-fiber composites are frequently used in dentistry. In order to evaluate their biocompatibility we tested, in an experimental model "in vivo", their tissue response pointing our attention on presence of mast cells (MCs) and fibrotic process. Sprague Dawley rats were used for the experimental design. The fibers were introduced in a subcutaneous pocket along the middle dorsal line between the two scapulas for 7, 14 or 21 days. At the end of the treatments the skins were excised and then processed for Toluidine Blue, to determine the presence of MCs, and Picrosirius Red staining, to evaluate the presence of fibrotic tissue. Our preliminary results showed and increase of both MC number and deposition of collagen type I, which characterized the fibrotic tissue. So, subsequent aims of our study were to evaluate the role played by MCs in tissue fibrosis and to give a possible explanation regarding the mechanisms that were responsible of biological response observed, through the analyses of some proteins, such as metalloproteinase-2 (MMP-2), its inhibitor (TIMP-2) and transforming growth factor-beta (TGF-beta). Our data confirmed the involvement of TGF-beta, released by MCs, in the disruption of the equilibrium between MMP-2 and TIMP-2 that were implicated in the enhancement of fibrosis. In summary, this study demonstrate that this type of materials induced an inflammatory response at the site of implant and help to clarify what type of mechanism and which proteins are involved in this biological response. Nevertheless, more extensive investigations are in progress to better evaluate the inflammatory process.

Caffeine (Caf) is largely used to delay fatigue, improving physical activity. However, its role remains elusive, and there are no hemodynamic or immunohistochemical data regarding its effects on skeletal muscle. We studied the hemodynamic and NOS expression of Bax/Bcl2 in skeletal muscle after single Caf administration. Thirty-two male rats were divided into six groups: the first was iv-injected with Caf (16 mg/kg), the second with Caf + L-NAME, the third with Caf + L-arg, the fourth with Caf + L-NAME + L-arg, fifth with saline. Mean arterial blood pressure (MAP) was monitored for 30′, then the animals were killed. The sixth group was injected with Caf and killed after 2 h. The quadriceps were isolated and processed by immunohistochemistry. We found that Caf increased MAP temporarily, while Caf + L-NAME increased it for a longer period. In untreated muscle, all NOS isoforms was expressed with different intensity and localisation, and Bcl2 was strongly expressed among the myofibrils. In Caf and Caf + L-NAME treated animals, NOS expression was lost; Bcl2 expression decreased among myofibrils but increased inside the subsarcolemma. The L-arg administration reversed these Caf and L-NAME effects. Two hours after Caf, NOS expression increased. We concluded that improved physical performance could be related to Caf's ability to interfere with the endogenous muscular synthesis of NO.

Aluminium (Al) impairs the glutamate-nitric oxide-cGMP pathway and reduces the number of nitroxidergic neurons in the rat somatosensory cortex. To understand better the effect of the time of exposure, we monitored the effect of aluminium administration on nitroxidergic neurons, identified by NADPH-diaphorase (NADPH-d) or by nitric oxide synthase (NOS) staining, after 0.5, 1, 2, 3, 6 and 12 months of aluminium administration. Since neuropeptide Y (NPY) is known to be colocalised with nitric oxide synthase in cortical neurons, the aim of this work was to study the effects of Al administration on the cortical expression of NADPH-d, nNOS, and NPY. NADPH-d or NOS positive neurons were found scattered in the cortex where they constituted about 1% of all neurons. Double staining using NADPH-d and NPY showed that almost all nitroxidergic neurons were co-localised with NPY neurons (NADPH-d/NPY double stained neurons) whereas some neurons were stained only with NPY (NPY single stained neurons) ; these were more numerous than NADPH-d/NPY double stained neurons. Al significantly reduced NADPH-d and nNOS positive neurons in the cerebral cortex time dependently, with the greatest effect appearing after 3 months. Also measured was the integrated optical density (IOD) of nNOS positive neurons showing a significant decrease of NOS immunostaining even in the remaining NOS positive neurons. The double staining experiment exhibited a decrease in NADPH-d/NPY double stained neurons with an apparent increase in NPY single stained neurons; these then decreased after 6-12 months. On the whole, the results confirm that Al impairs nitroxidergic pathways time dependently; moreover, the transient increase in NPY single stained neurons from 1 to 3 months suggests that there is an intraneuronal down-regulation of NOS, without affecting neuronal viability. In addition, the decrease in the NPY system found at 6 and 12 months may indicate that Al affected nitroxidergic and NPY systems at different times.

The extra-cellular matrix of the gingival tissue plays an important role in the homeostasis of dental implants. In this work we have studied immunohistochemically the distribution of collagen I-III-IV-V, tenascin, metalloproteinases (MMP) 1-3-8-13 and TIMP-1 in three groups of patients: (1) subjects with natural teeth (healthy periodontal tissue), (2) subjects with normal peri-implant mucosa and (3) subjects with clinically evident peri-implantitis. The immunolabelling for collagen I-III-IV showed a similar pattern in all three groups. The labelling for collagen V increased in lamina propria of healthy peri-implant tissue and peri-implantitis. Tenascin immunolabelling in healthy and peri-implant tissues was scattered in lamina propria. In peri-implantitis tenascin immunolabelling increased mainly near to the basal lamina. The MMP-1-3-8 and TIMP-1 immunolabelling were very faint and localized in the stroma in all three groups. In healthy and peri-implant tissues MMP-13 immunolabelling was found in the lamina propria whereas in peri-implantitis MMP-13 immunolabelling was also in epithelium. On the whole, these data suggest that in the extracellular matrix of peri-implantitis there are alterations of collagen V, tenascin and MMP-13 patterns.

Cyclosporine A (CsA) use is associated with several side effects, the most important of which is nephrotoxicity that includes, as we previously showed, tubular injury and interstitial fibrosis. Recently, many researchers have been interested in minimizing these effects by pharmacological interventions. To do this, we tested whether the administration of a red wine polyphenol, Provinol (PV), prevents the development of CsA-induced nephrotoxicity. Rats were treated for 21 days and divided into four groups: control; group treated with PV (40 mg/kg/day by oral administration in tap water); group treated with CsA (15 mg/kg/day by subcutaneous injection); group treated with CsA plus PV. CsA produced a significant increase of systolic blood pressure; it did not affect urinary output, but caused a significant decrease in creatinine clearance. These side effects were associated with an increase in conjugated dienes, which are lipid peroxidation products, inducible NO-synthase (iNOS), and nuclear factor (NF)-kB, which are involved in antioxidant damage. However, PV prevented these negative effects through a protective mechanism that involved reduction of both oxidative stress and increased iNOS and NF-kB expression induced by CsA. These results provide a pharmacological basis for the beneficial effects of plant-derived polyphenols against CsA-induced renal damage associated with CsA.

Cyclosporine A (CsA) is the immunosuppressant of first choice in allotransplantation. Its use is associated with side effects of nephrotoxicity and neurotoxicity, which are among the most prominent. This study was undertaken to explore whether expression and activity of heme oxygenase (HO), the rate-limiting enzyme in heme degradation, is altered in a rat model of CsA-induced injury. Male Sprague Dawley rats were divided into four groups and treated for 21 days. Group I (control) was injected with olive oil (vehicle), group II with CsA (15 mg/kg/day), group III with CsA and the HO inhibitor stannous mesoporphyrin (SnMP) (30 micromol/kg/day) and group IV with one dose of the HO inducer cobalt protoporphyrin (CoPP) 5 mg/100 or heme (10 mg/kg body weight), three days after onset of CsA treatment. Renal tissue was processed for light microscopy, and for HO-1 enzyme activity, assay and for Western blot analysis. In CsA-treated rats there was histological evidence of tubulointerstitial scarring. HO-1 was undetectable in CsA-treated rats compared to control while there was no change in HO-2. In animals treated with a combination of CsA and SnMP, HO-1 activity was further reduced. In animals treated with a combination of CsA and CoPP, HO-1 protein levels were partially restored. These observations indicate that downregulation of HO-1 expression by CsA could be one mechanism underlying CsA-induced toxicity. The CsA-induced decrease in HO-1 expression is partial and restorable, and attempts to preserve HO levels may attenuate CsA toxicity.

SUMMARY Cyclosporine A (CsA) is the first immunosuppressant used in allotransplantation. Its use is associated with side effects that include nephrotoxicity. This study explored the anatomic structures involved in CsA nephrotoxicity and the effect of heme oxygenase (HO) in preventing CsA injury. Rats were divided into four groups, which were treated with olive oil, CsA (15 mg/kg/day), CsA plus the HO inhibitor (SnMP; 30 � M/kg/day), and with the HO inducer (CoPP; 5 mg/100 g bw). Renal tissue was treated for morphological, biochemical, and immunohistochemical studies. CsA-treated rats showed degenerative changes with renal fibrosis localized mainly around proximal tubules. Collapsed vessels were sometimes seen in glomeruli. No HO-1 expression and increased expression of endothelin-1 (ET-1) were observed in CsA-treated rats compared with controls. In CsA plus SnMP-treated rats, HO-1 expression was further reduced and the morphology was not changed compared to the CsA group, whereas CsA plus CoPP-treated animals again showed normal morphology and with restoration and an increase in HO-1 levels. HO activity and immunohistochemical data showed similar alterations as HO expression. No changes were observed for HO-2 analysis. The observations indicate that HO-1 downregulation and ET-1 upregulation by CsA might be one mechanism underlying CsA-induced nephrotoxicity. Therefore, attempts to preserve HO levels attenuate CsA nephrotoxicity. (J Histochem Cytochem 53:105–112, 2005)

The immunosuppressive drug Cyclosporine A (CsA) has been successfully used in several diseases with immunological basis and in transplant patients. However, the therapeutic treatment induces several side effects, which include the development of severe hypertension, renal failure and cardiotoxicity in the majority of the patients. Since the mechanism by which CsA induces hypertension is not well defined, the aim of this study is to evaluate the morphological changes and the expression of heat shock proteins (HSPs) in the thoracic aorta of CsA-treated rats. The study was carried out on 40 male Wistar rats with an average weight of 200-250 g. The animals were divided into four groups. Groups I and II were injected subcutaneously (sc) daily with castor oil for 15 or 30 days and used as control; group III and IV were injected sc daily with CsA (15 mg/Kg/day) for 15 or 30 days. After the end of the treatment, the thoracic aortae were removed and treated for morphological (Sirius Red) and immunohistochemical evaluation (HSP 25, alphaB-crystallin and HSP 47). The results indicate that CsA induces (1) time-dependent vascular damage visible as fibrosis mainly in intima-media tunica of aorta, and (2) a clear increase in HSP expression. In fact, after 30 days of treatment, HSP and alphaB-crystallin are increased in all tunicas, whereas HSP47 only in tunica media and adventitia. These findings could suggest that these proteins are up-regulated after CsA treatment in order to play a defensive role in vascular damage.

Vigorous inflammatory responses are associated with tissue damage, particularly when toxic levels of inflammatory cytokines are produced. Despite proangiogenic factors being present early at sites of inflammation, vascular repair occurs toward the end of the inflammatory response, suggesting modulation of the proangiogenic response. Endogenous inhibitors of angiogenesis induced during acute inflammation are poorly characterized. Here, we looked for endothelial cell-derived modulators of angiogenesis that may account for delayed neovascularization during inflammation. Gene profiling of endothelial cells showed that the inflammatory cytokines tumor necrosis factor α (TNF-α) and interferon-γ (IFN-γ) selectively promote expression of the antiangiogenic molecules, IFN-inducible protein-10, monokine induced by IFN-γ, tryptophanyl-tRNA synthetase, and tissue inhibitor of metalmetalloproteinase-1, and inhibit expression of the proangiogenic molecules, platelet-endothelial cell adhesion molecule-1, vascular endothelial growth factor receptor-2, stromal cell-derived factor-1 (SDF-1), collagen type IV, endothelial cell growth factor-1, and carcinoembryonic antigen-related cell adhesion molecule-1. Reduced endothelial cell expression of SDF-1 protein by TNF-α and IFN-γ disrupts extracellular matrix-dependent endothelial cell tube formation, an in vitro morphogenic process that recapitulates critical steps in angiogenesis. Replacement of SDF-1 onto the endothelial cell surface reconstitutes this morphogenic process. In vivo, TNF-α and IFN-γ inhibit growth factor-induced angiogenesis and SDF-1 expression in endothelial cells. These results demonstrate that SDF-1/CXC chemokine receptor-4 constitutes a TNF-α- and IFN-γ-regulated signaling system that plays a critical role in mediating angiogenesis inhibition by these inflammatory cytokines.

Cyclosporine A (CsA) is the first immunosuppressant universally used in allotransplantation. However, it has been demonstrated that this drug produces side effects in several organs, particularly in the kidney and in the heart. Since the immunosuppressive therapy induces myocardial toxicity, the aim of this study was to evaluate in the myocardium of CsA-treated rats the expression variations of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 2 (MMP2), to verify if: VEGF increased, VEGF increase was associated with MMP2 increase and they could be considered as repair proteins. The study was carried out on 28 Wistar rats divided into two groups. The group I animals served as control and so they were subcutaneously injected daily with castor oil for 21 days; group II: animals were subcutaneously injected daily with CsA (Sandimmun, Sandoz) at the therapeutic dose (15 mg/kg) for 21 days. The group I animals (control) showed normal heart architecture and low levels of MMP2 and VEGF. The group II animals (CsA-treated) showed structural degenerative changes with myocardial fibrosis and a clear increase both in MMP2 and VEGF. These data show that the immunosuppressant therapy induces a high increase in both the proteins and that VEGF variations are associated with MMP2 variations. These findings suggest that these proteins are involved in the endogenous emergency cell mechanisms induced by stress conditions, probably playing a role against myocardial injury as repair proteins.

We have evaluated the effects of an ACE inhibitor, enalapril (ENA) and of an angiotensin II receptor blocker, losartan (LOS), administered either at hypotensive or non-hypotensive dosage, on the cardiac and renal structure of spontaneously hypertensive rats (SHR). Forty-eight rats were included in the study: eight SHR were treated with low-dose (ld, 1 mg/kg/day) ENA; eight with low-dose (ld, 0.5 mg/kg/day) LOS; eight with high-dose (hd, 25 mg/kg/day) ENA; eight with high-dose (hd, 15 mg/kg/day) LOS; while eight Wistar-Kyoto (WKY) and eight SHR were kept untreated (unt). Treatment was given from the 4th to the 12th week of age. Systolic blood pressure (SBP) was measured non-invasively every week. The left ventricular weight to body weight (RLVM) and the left + right kidney weight (RKW) to body weight was measured, and the cardiac and glomerular interstitial collagen content was evaluated using sirius red staining and image analysis. In addition, cardiac metalloproteinases activity (43 kDa MMP, MMP-2, and MMP-9) was evaluated by zymography. A significant reduction in RLVM was observed in SHR given ENA hd or LOS hd. Cardiac collagen was significantly reduced in SHR ENA hd and SHR LOS hd as well as in SHR LOS ld, but not in SHR ENA ld. The 43 kDa MMP collagenase activity was greater in WKY unt compared with SHR unt, being normalized only in SHR ENA hd. The gelatinase activity of MMP-9 showed a trend similar to 43 kDa MMP, but differences between SHR and WKY unt were only of borderline statistical significance. No difference among groups was observed in MMP-2 activity. No significant differences in RKW was observed between groups. However, the collagen content in the glomerular perivascular space was significantly reduced in all treated groups, including those given ld, compared with SHR unt. In conclusion, LOS and ENA showed a similar preventive effect on the increase of RLVM in SHR, but, at least in part, different effects on the extracellular matrix in different organs, being cardiac collagen less sensitive to low dose (ld) ACE inhibition.

We have evaluated the expression of metabotropic glutamate receptors (mGluR subtypes 2/3, 4 and 5) in rat thymus under normal and experimental conditions after 2 and 21 days of cyclosporine-A treatment. In normal rats, immunohistochemical analysis showed that expression of mGluRs was high in dendritic cells and lymphocytes of the medulla whereas it was weak in lymphocytes of the cortex. However, there were some differences in the expression of mGluRs subtypes. mGluR5 showed strong expression in lymphocytes of medulla and dendritic cells. mGluR2/3 and mGluR4 were moderately expressed in lymphocytes and dendritic cells of the medulla and weakly in cortical lymphocytes. Immunoblotting showed moderate levels of mGluR2/3 and mGluR4 and strong levels of mGluRS. After 2 days of cyclosporine-A treatment, we observed by immunohistochemistry and immunoblotting a distinct decrease in all mGluRs and their expression had almost completely disappeared after 21 days of treatment. The results clearly indicate that: 1) mGluR2/3, 4 and 5 are widely expressed in thymic cells; 2) the mGluR5 subtype is expressed most strongly in medullary cells; and 3) cyclosporine-A rapidly inhibits expression of all mGluR subtypes after 2 days of treatment and their complete disappearance after prolonged treatment. These findings may indicate a possible mechanism by which cyclosporine-A produces its immunosupressive effects.

Renal damage caused by therapeutic treatment with cyclosporine A has been well documented. Clinical experiences have shown that cyclosporine A nephrotoxicity is determined by interstitial fibrosis with tubular atrophy. However, the exact mechanism by which this drug causes nephrotoxicity has not yet been clarified. This study used an in vitro model in an attempt to identify the cellular mechanisms underlying kidney cyclosporine A damage. We used two cell lines with the characteristics of proximal and distal tubule cells (pig kidney proximal tubular epithelial cell line [LLC-PK1] and Madin-Darby canine kidney cell line [MDCK]. The cell lines were treated with cyclosporine A for 24 h. After the treatment, the cells were stained with Trypan Blue to estimate cell viability and processed by histochemical reactions to evaluate their cellular metabolism. Four enzymes (acid phosphatase, alkaline phosphatase, lactate dehydrogenase and succinate dehydrogenase) were considered. The cell viability assay showed that the LLC-PK1 cell line was more sensitive to cyclosporine A than MDCK. Remarkably, the LLC-PK1 cells disappeared with cyclosporine A treatment. As for the hydrolytic enzymes, only acid phosphatases showed an increased positivity in the treated LLC-PK1 cells. Similarly, lactate dehydrogenase showed a different activity histochemically. No statistically significant alterations were observed in the succinate dehydrogenase reaction. The cyclosporine A-treated MDCK cell lines did not show any difference in either their hydrolytic or succinate dehydrogenase enzyme positivity with respect to the control line. In contrast, there was a significant increase in lactate dehydrogenase activity. This study allowed the possible mechanism of cyclosporine A-induced damage in renal tubular cells to be evaluated. The enzymatic changes happened rapidly (during the 24 h of treatment), suggesting that this alteration was one of the steps by which cyclosporine A induced toxicity. Moreover, since acid phosphatase is a marker of protein catabolism, the variation in the activity of this enzyme, in the LLC-PK1 line only, showed that cyclosporine can induce alterations leading to cellular toxicity. The modifications in lactate dehydrogenase activity, in both lines, suggested that this drug caused cell stress, inducing the production of lactic acid from glucose in the presence of oxygen. In conclusion, cyclosporine A treatment may force LLC-PK1 and MDCK cells to use anaerobic glycolysis preferentially. Further, these enzyme alterations may represent an epiphenomenon or a consequence of cyclosporine A toxicity.

The effect of melatonin on experimental prolonged cyclosporine A (CsA) nephrotoxicity was analyzed by electron microscopy and morphometry. Twenty female Wistar rats were divided into groups that received (1) CsA at a therapeutic dose (15 mg/kg/day) sc for 40 days; (2) olive oil, CsA vehicle, as controls; (3) CsA plus melaton (1 mg/kg) ip for the same time; (4) melatonin alone, as additional controls. All controls had normal renal ultrastructure. However, in CsA group, both tubular and glomerular alterations were observed. The authors found apoptotic and necrotic proximal tubules with disrupted brush border, swollen mitochondria, abundant lysosomes; in the glomerulus, amorphous basement membrane, and abnormal mesangial matrix. CsA plus melatonin administration partly prevented these changes. Nevertheless, fibrillar deposits in podocytes and basolateral membrane dilatations in proximal tubules were observed. Histopathological analysis on semithin sections and ultrastructural morphometry were also performed. CsA induced interstitial fibrosis, atrophy, and PAS positive inclusions in the proximal tubules. Moreover, CsA reduced glomerular and mesangial volume but enhanced mesangial matrix volume and basement membrane thickness. After CsA plus melatonin, these parameters were reduced in the proximal tubules and restored in the glomerulus. In conclusion, melatonin attenuated morpho-quantitative alterations induced by CsA in the rat kidney.

Small heat shock proteins (sHSPs) are cytoskeletal chaperones constitutively expressed in the normal kidney but enhanced with beneficial roles during adverse stimuli. Cyclosporine A is an immunosuppressive drug with major adverse side effect such as severe nephrotoxicity. Among possible mechanisms of cyclosporine A-induced renal damage, oxidative stress and cytoskeletal damage have been suggested. Melatonin has been successfully used as antioxidant against many renal diseases. This in vivo study was performed to shed light on the protective effect of melatonin against cyclosporine A-induced renal alterations. We treated rats with cyclosporine A alone, or combined with melatonin, and with melatonin alone (as controls) for 40 days and analysed the renal abundance and distribution of two sHSPs, HSP25 and alpha B-crystallin. These data were correlated with the histopathological effects of the treatments. Cyclosporine A induced insoluble isoforms that moved to soluble fractions after melatonin coadministration as in controls. After cyclosporine A treatment, an intense signal for sHSPs was found within the glomeruli, nucleus and cytoplasm of cortical tubules, collecting ducts and vascular wall. After melatonin supply, the staining was faint, limited to the cytoplasm of cortical tubules, similar to controls. Both fibrosis and tubular alterations significantly decreased after melatonin coadministration. In conclusion, HSP25 and alpha B-crystallin are overexpressed in the rat kidney treated with cyclosporine A but are similar to controls after combined melatonin. This could be a consequence of the cytoprotective effect of melatonin in this nephrotoxic model so that a beneficial sHSPs response isbreak unnecessary.

Pro-inflammatory cytokines play a key role in the inflammatory response and contributing to producing local hyperalgesia. It is known that pentoxifylline inhibits inflammatory cytokine production. Therefore, the aim of this study was to find out, using a behavioural test and neuronal Fos expression, if this drug had some antinociceptive effect decreasing nociresponsive neuron activation in the spinal cord after nociceptive inflammation (injection of formalin in the hind paw). Our data showed that formalin induced a great number of Fos-positive neurons in the dorsal horn of the lumbar spinal cord mainly in the ipsilateral I and II laminae. The local pre-treatment with pentoxifylline before the formalin injection, decreased the nociceptive response in the formalin test and the number of Fos positive neurons in the spinal cord. This finding suggests that pentoxifylline is involved in decreasing the activation of nociceptive neurons in the spinal cord playing a role in local anti-nociception.

No medical or surgial treatments have been reported to permanently stop trigeminal pain attacks. TEMPOL, a stable nitroxide radical scavenger, has been recently shown efficient in providing neuroprotection. Here we tested the efficiency of TEMPOL in alleviating heat hyperalgesia and expression of NADPH-d in trigeminal afferent neurons of rats with experimentally induced secondary trigeminal neuralgia by chronic constriction of infraorbitary nerve. The results showed that TEMPOL significantly reduced heat hyperalgesia and induced a large decrease of NADPH-d in neurons of the Gasser ganglia. These findings suggest that the radical scavengers like TEMPOL could be new therapeutic strategies to treat neuropathic pain induced by infraorbital nerve constriction.

Background —It is not presently known whether non–insulin-dependent diabetes mellitus (NIDDM) is associated with the presence of structural alterations in small arteries or whether the combination of hypertension and NIDDM may have an additive effect on endothelial dysfunction. Therefore, we investigated subcutaneous small arteries in 12 normotensive subjects (NT group), 18 patients with essential hypertension (EH group), 13 patients with NIDDM, and 11 patients with NIDDM and EH (NIDDM+EH group). Methods and Results —Subcutaneous small arteries were evaluated by a micromyographic technique. The internal diameter, the media-to-lumen ratio, remodeling and growth indices, and the collagen-to-elastin ratio were calculated. Concentration-response curves to acetylcholine, bradykinin, the endothelium-independent vasodilator sodium nitroprusside, and endothelin-1 were performed. The media-to-lumen ratio was higher in the EH, NIDDM, and NIDDM+EH groups compared with the NT group. EH patients showed the presence of eutrophic remodeling, whereas NIDDM and NIDDM+EH patients showed 40% to 46% cell growth. The collagen-to-elastin ratio was significantly increased in the EH and NIDDM+EH groups compared with the NT group. The vasodilatation to acetylcholine and bradykinin was similarly reduced in EH, NIDDM, and NIDDM+EH groups compared with the NT group. The contractile responses to endothelin-1 were similarly reduced in EH, NIDDM, and NIDDM+EH patients. Conclusions —Our data suggest that the effects of NIDDM and EH on small artery morphology are quantitatively similar but qualitatively different and that the presence of hypertension in diabetic patients has little additive effect on small artery morphology and none on endothelial dysfunction.

In the present study, we investigated the variation of endothelin (ET) levels in various organs (kidney, liver, spleen and heart) of rats after 7 and 15 days of Cyclosporine A (CsA) treatment. Moreover, we studied whether these modifications are related to major toxic effects, known to be a complication of CsA treatment (i. e. nephrotoxicity). The morphology of liver, spleen and heart in rats treated with CsA for 7 and 15 days was similar to that in control rats. The amounts of ET protein as determined by Western blot analysis were very low in all organs. However, we found lesions in kidneys of rats treated with CsA for 15 days but not in kidneys of rats treated for 7 days. Immunohistochemical analysis using an ET polyclonal antibody revealed that proximal tubules of animals treated for 15 days were strongly ET positive whereas distal tubules and glomerula showed only weak positivity as those in control rats. Western blot analysis revealed an increase in ET protein in treated rats. On the basis of these data, we conclude that CsA induced evident nephrotoxicity already after 15 days of treatment and that toxic effects of CsA were related to the amounts of ET found. An explanation for these findings is that ET, produced by epithelial cells, is filtered through the glomerular tuft and resorbed by tubular epithelial cells. Since variation in levels of ET was visible only in proximal tubules, we conclude that CsA treatment during a brief period produced side effects that can be considered as acute toxicity. Our finding may help to understand the mechanism of CsA toxicity and may provide important clues for pharmacological strategies to reduce CsA toxicity.

We have evaluated whether cyclosporine A affects cell structure and cytoskeletal proteins of the thymus of Wistar rats. Immunohistochemical analysis showed that expression of the cytoskeletal proteins vimentin and desmin was much higher in epithelial cells, dendritic cells and lymphocytes in the thymus of treated rats than in untreated controls. Protein expression was observed as a positive condensation in a distinct area near the nucleus with a capping-like configuration. An ultrastructural study showed that the amount of cytoskeletal fibrillar structures was increased in the treated rats. The structures were assembled in a limited area of the cell with a nuclear capping-like configuration which was in agreement with the light microscopical observations. Immunoblotting analysis demonstrated that vimentin and desmin had a lower molecular weight in treated rats than in controls (57 and 53 kDa versus 55 and 51 kDa, respectively). The results clearly indicate that cyclosporine A affects the structure of the cytoskeleton suggesting that this could be the first step in its immunosuppressive effects by altering nucleus/cytoplasm signaling.

Nitric oxide (NO) is a neuronal messenger that it is thought to be involved in the nociceptive transmission modulation. The activity of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) was shown to be identical to NOS activity in the brain. Since the periaqueductal gray matter (PAG) plays an important role in pain perception and antinociception this study was carried out to monitor the expression of NADPH-d in PAG after nociceptive visceral stimulation. Our data showed that the noxious visceral stimulation significantly increased NADPH-d positive neurons and that these neurons were localized in the ventrolateral areas of the PAG. These findings suggest that NO in the PAG may play a role in pain modulation and antinociception.

A case of severe cardiac involvement is reported in a patient affected with familial amyloidotic polyneuropathy due to the Portuguese type I variant (Val→Met30) of the transthyretin (prealbumin) molec

This paper intends to focus on the transition from a “productivist” and farm business development model - dominant until the 1970s within the framework of the Common Agricultural Policy (CAP) - to a multifunctional, environmentally sustainable and integrated diversified, alternative, and locally sustainable productive perspective. In the 1980s, this model evolved by means of a fundamental, and principally cultural, development. It has shaped rural development policies, marking the down-sizing of old price and market policy. First of all, the intention is to sketch a brief evolutionary-historical outline of the CAP orientation; agritourism is then identified as a product innovation a la Schumpeter and an interesting diversification of farm productive activities. Agritourism is analysed from a multifunctional perspective, aimed at safeguarding the integrity of natural resources and also at enhancing socio-cultural aspects and the oeno-gastronomical heritage of specific rural areas. There is also a brief review of agritourism, present in the Italian juridical system from the 1980s, seen from the perspective of a researcher into economic theory: attention is focused specifically on Law No. 96/2006. Zusammenfassung Dieser Beitrag fokussiert auf den Ubergang von dem in der Gemeinsamen Agrarpolitik der EU in den 1970er Jahren vorherrschenden produktivitatsorientierten Modell der landwirtschaftlichen Betriebsentwicklung zu einem Paradigma der multifunktionalen, okologisch nachhaltigen Landwirtschaft. Der Primarsektor wird hierbei als Teil des landlichen Raums mit einem systemischen, diversifizierten und lokal nachhaltigen Fokus konzipiert. Die so veranderte landliche Entwicklungspolitik ist durch die Ruckfuhrung der klassischen Instrumente der Markt- und Preispolitik gekennzeichnet. Im Folgenden soll zunachst ein kurzer Uberblick uber die Entwicklung der Ausrichtung der GAP im Zeitablauf gegeben werden; daraufhin wird der Agrartourismus als eine Schumpeterianische Produktinnovation und interessante Diversifikationsstrategie fur landwirtschaftliche Betriebe beschrieben. Dabei wird der Landtourismus in multifunktionaler Hinsicht analysiert, der zur Erhaltung naturlicher Ressourcen ebenso wie zur Starkung soziokultureller Aspekte und des onogastronomischen Erbes spezifischer landlicher Regionen beitragt. Daruber hinaus wird ein kurzer Uberblick uber den Agrartourismus, der seit den 1980er Jahren im italienischen Rechtssystem verankert ist, aus Sicht der okonomischen Theorie gegeben. Der spezielle Fokus liegt dabei auf dem Gesetz Nr. 96/2006.

The effects of somatostatin-14 (S) on the proliferation of the human prostatic cancer cell line (LNCaP, lymph node carcinoma of the prostate) and on the amount of proteins secreted by these cells have been studied. LNCaP cells were treated for 3 days with different doses of S. The hormone significantly inhibits cell proliferation at doses comprised between 0.4 and 2 nmol, as indicated by a significant decrease in the incorporation of 3H-thymidine as well as in the uptake of [alpha-32P]dATP. The direct antiproliferative effect of S is reversible, since cell proliferation returns to normal 4 days after withdrawal of the hormone from the medium. This action of S is not due to cytotoxic side-effects, since no changes in the incorporation of 35S-methionine are observed. Total RNA and protein synthesis do not appear to be modified by the hormone in vitro. The hormone probably exerts its antiproliferative effect on LNCaP cells by stimulating phosphotyrosyl protein phosphatases, since the inhibition of growth induced by S (1 nmol) is reversed by sodium vanadate (2 and 4 mumol), a potent inhibitor of the process of tyrosine dephosphorilation. In addition, S (1 nmol/L) induces a significant decrease in the amounts of proteins secreted by LNCaP cells. Also the antisecretory action of S seems to be mediated by the activation of phosphotyrosyl protein phosphatases, since sodium vanadate is able to reverse the action of S on this parameter. In conclusion, the present data suggest for the first time that S exerts a direct inhibitory effect on LNCaP cell proliferation and protein secretion, two effects probably mediated by the activation of phosphotyrosyl protein phosphatases.

International audience; Flavonoids, polyphenol derivatives of plant origin, possess a broad range of pharmacological properties. A number of studies have found both pro/anti-apoptotic effects for many of these compounds. For these reasons we investigated whether Provinols flavonoids obtained from red wine, have anti-apoptotic properties. The investigations have been carried out in rats treated with Cyclosporine A (CsA). In particular, four groups of rats have been treated for 21 days with either olive oil (control group), with CsA, with Provinols, or with CsA and Provinols simultaneously. Oxidative stress, systolic blood pressure, body weight, biochemical parameters and different markers of pro/anti-apoptotic pathway were measured. CsA produced an increase of systolic blood pressure, a decrease in body weight, serum creatinine levels, urinary total protein concentration and creatinine clearance. Moreover, CsA induced renal alterations and the translocation of Bax and cytochrome c from cytoplasm to mitochondria and vice versa. These changes activated the caspase cascade pathway, that leads to morphological and biochemical features of apoptosis. Provinols restored morphological and biochemical alterations and prevented nephrotoxicity. In conclusion, this study may augment our current understanding of the controversial pro-/anti-apoptotic properties of flavonoids and their molecular mechanisms.

A morphoquantitative study was carried out to clarify the cytoarchitecture of the human nuclear complex of the posterior commissure (NPC), a collection of cell groups which is thought to be involved in the pupillary light reflex and eye movements. On the basis of the topographical features, 5 different parts (rostral, principal, magnocellular, subcommissural and infracommissural) were identified in NPC. In these 5 parts, the neuronal population consisted of small and medium-sized neurons with a structural pattern similar to that of the central reticular formation. The only exception was the magnocellular part in which the neurons, mainly medium-sized and large, did not show the reticular-like features. In the Golgi material, two types of NPC neurons were identified: multipolar and fusiform cells. In the multipolar cells, 2-3 dendrites emerged from the soma and gave off a wide dendritic arborization. Many of the dendrites and axons of the multipolar cells were seen spreading outside NPC. The fusiform cells had 2 dendrites emerging from the opposite poles of their elongated perikaryon. Dendrites and axons of the fusiform cells always lay inside NPC. These observations suggest that the multipolar cells are mainly projective neurons and that the fusiform cells are neurons which mainly perform a local integrative function. Since the present study concludes our investigation of the human accessory oculomotor nuclei, a comparison of their structure was carried out. All the nuclei showed a common reticular-like structure, although each nucleus had its own structural particularities. These results are in agreement with and give an anatomical support to the functional findings.

Aging is associated with progressive structural disorganization of muscular and cardiac fibers, decreasing functional capacity, and increased rates of disease and death. Aging is also characterized by disturbances in protein synthesis with impaired cellular organelle functions, particularly in the mitochondria. The availability of amino acids is a key factor for the overall metabolism of mammals and exogenous supplements of amino acid mixtures (AAm) could be a valid therapeutic strategy to improve quality of life, avoiding malnutrition and muscle wasting in the elderly. We investigated the morphoquantitative effects of long-term AAm supplementation on the mitochondria and sarcomeres (by electron microscope) and on collagen matrix deposition (by histologic techniques) in both skeletal and cardiac muscles of young and aged mice. Our data showed that old animals have fewer mitochondria and massive fibrosis in both muscles. Long-term AAm supplementation increased the number and volume of mitochondria and sarcomeres and decreased fibrosis in both skeletal muscle and hearts in old rats. These findings indicate that AAm restored muscular morphologic parameters and probably improved the mechanical performance of these organs.

Aluminium (Al) is a ubiquitous metal that is potentially toxic to the brain. Its effects on other fundamental organs are not completely understood. This morphological in vivo study sought to compare sublethal hepatotoxic changes and Al deposition in adult mice that orally ingested Al sulphate daily for 10 months, in age matched control mice that drank tap water and in senescent mice (24 months old). Livers were examined for collagen deposition using Sirius red and Masson, for iron accumulation using Perls' stain. Light, electron microscopy and morphometry were used to assess fibrosis and vascular changes. Scanning transmission electron microscopy and EDX microanalysis were used to detect in situ elemental Al. Iron deposition, transferrin receptor expression were significantly altered following Al exposure and in the aged liver but were unaffected in age matched control mice. In Al treated mice as in senescent mice, endothelial thickness was increased and porosity was decreased like perisinusoidal actin. Furthermore, Al stimulated the deposition of collagen and laminin, mainly in acinar zones 1 and 3. Pseudocapillarization and periportal laminin in senescent mice were similar to Al treated adult liver. In conclusion, prolonged Al sulphate intake accelerates features of senescence in the adult mice liver. (C) 2008 Elsevier Inc. All rights reserved.

Substance P (SP) is a non-opioid peptide that generates a potent, analgesia when injected into the periaqueductal gray matter (PAG)The aim of this study was to investigate the fine neuronal structures and synaptic circuits involved in SP action in rats by means of electron microscopy, using immunocytochemical (ICC) pre-embedding methods. A conventional ultrastructural study, carried out to interpret the ICC data correctly, shows small sized nerve cell bodies with a high nucleus–cytoplasmic ratio; absence of an extensive granular endoplasmic reticulum; and few axo-somatic contacts having symmetrical and asymmetrical junctions in equal proportions. The large neuropil is characterized by numerous thin unmyelinated axons and axodendritic synapses mainly showing pleomorphic vesicles and asymmetrical junctions. The ICC analysis showed moderately labeled nerve cell bodies with the same structural, synaptic, and dimensional features as the negative cells. In the neuropil SP immunoreactivity is shown by dendrites, synapses, and thin elements which are unidentifiable structurally. No SP terminals synapsing on SP nerve cell bodies were found and only occasional SP light labeled terminals synapsing on negative perikarya were seen. The SP boutons generally have pleomorphic vesicles and asymmetrical junctions. On the basis of these data a possible excitatory activity of PAG SP synapses could be hypothesized. This activity would take place on postsynaptic neurons generally at a dendritic level. Our ultrastructural findings give support to an excitatory role carried out by SP neurons of the PAG, as suggested by the role of PAG circuitry on spinal nociception.

Caffeine is the most frequently ingested neuroactive drug in the world and it is largely used to delay fatigue and improve physical activity. Caffeine can modulate NO synthesis in cells and may influence muscular function by modifying the cellular cycle life-death. There is little data concerning the relationship between caffeine in the heart, NOS expression and apoptosis and no data regarding the acute effect of high doses of caffeine in the in vivo myocardium. We therefore studied hemodynamic NOS and Bax/Bcl2 expression in the rat myocardium after a single cafffeine administration. Thirty-two male rats were divided into six groups: the first was iv-injected with caffeine (16 mg/kg), the second with caffeine + l-NAME (30 mg/kg), the third with caffeine + l-arg (0.5 g/kg), the fourth with caffeine + l-NAME + l-arg and finally the fifth with saline. Mean arterial blood pressure (MAP) was monitored for 30 min, then the animals were killed. The sixth group was injected with caffeine and killed after 2 h. The hearts were isolated and processed by immunohistochemistry. We found that caffeine increased MAP temporarily while caffeine + l-NAME increased it for a longer period. In the control myocardium, all NOS isoforms were expressed. The Bcl2 were strongly expressed inside the perinuclear cytoplasm whereas Bax was very faintly detectable in the peripheral cytoplasm. In caffeine and caffeine + l-NAME treated animals, NOS expression disappeared. Bax and Bcl2 expression did not vary. The l-arg administration reversed these caffeine and l-NAME effects on NOS expression. Two hours after caffeine, NOS expression increased and Bax and Bcl2 expression did not vary, although Bcl2 was mainly expressed in the peripheral cytoplasm. We conclude that improved caffeine-induced physical performance could also be related to caffeine’s ability to interfere with endogenous myocardial NO synthesis. Furthermore, we suggest that myocardial cell plays an effective anti-apoptotic role against acute caffeine administration. © 2007 Published by Elsevier Ltd.

The aim of the present study was to analyze the early events in atherogenesis and the role of pro- or anti-atherosclerotic proteins in the development of atherosclerotic lesions. We used apolipoprotein E-deficient (E(0)) mice that spontaneously develop hypercholesterolemia and atherosclerotic lesions in the aorta in a time-dependent manner. Aortas of mice aged 6, 8, 10 and 12 weeks were examined to determine histopathological changes. In mice aged 8-12 weeks, developing atherosclerotic lesions were present in different regions of the aortas. These lesions protruded into the lumen of the vessel and showed lipid deposits, lipid-filled macrophages and extensive accumulation of collagen and elastic fibers throughout the entire arterial wall. A parallel immunohistochemical study included analysis of three proteins known to be involved in atherosclerosis, i.e. inducible nitric oxide synthase (iNOS, NOS2), vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP2). Increased immunolabelling of iNOS and VEGF accompanied atherosclerosis development in E(0) mice aged 8, 10 and 12 weeks. On the contrary, immunolabelling for MMP2 was negative in E(0) mice aged 10 and 12 weeks. Our results indicate morphological alterations in the Tunica intima and Tunica media of atherosclerotic aortas and possible protective roles for iNOS and VEGF proteins against atherosclerosis development. These data may be relevant for developing therapeutic strategies for atherosclerosis development.

Cannabinoids, such as anandamide, are involved in pain transmission. We evaluated the effects of AM404 (N-(4-hydroxyphenyl)-5Z,8Z,11Z,14Z-eicosatetraenamide), an anandamide reuptake inhibitor, monitoring the expression of c-fos, a marker of activated neurons and the pain-related behaviours using formalin test. The study was carried out in an experimental model of inflammatory pain made by a single injection of formalin in rat hind paws. Formalin test showed that the antinociceptive effect of AM404 was evident in phase I. We found that Fos-positive neurons in dorsal superficial and deep laminae of the lumbar spinal cord increased in formalin-injected animals and that AM404 significantly reduced Fos induction. Co-administration of cannabinoid CB1 receptor antagonist (AM251), cannabinoid CB2 receptor antagonist (AM630) and transient receptor potential vanilloid type 1 (TRPV-1) antagonist (capsazepine), attenuate the inhibitory effect of AM404 and this effect was higher using cannabinoid CB2 and vanilloid TRPV-1 receptor antagonists. These results suggest that AM404 could be a useful drug to reduce inflammatory pain in our experimental model and that cannabinoid CB2 receptor and vanilloid TRPV-1 receptor, and to a lesser extent, the cannabinoid CB1 receptor are involved.

It has been suggested that heat-shock proteins (HSPs) might be involved in autoimmune disease mechanisms in humans, considering the high degree of sequence homology between bacterial and human HSPs. Several authors have postulated that HSPs might be involved in periodontal disease processes, but not specifically in peri-implantitis. Consequently, using immunohistochemical techniques, we studied the distribution of HSP25, HSP32, HSP60 and HSP72 in three groups of patients: (1) subjects with natural teeth (healthy periodontal tissue), (2) subjects with normal peri-implant mucosa and (3) subjects with clinically evident peri-implantitis. The immunolabelling for HSP25 and HSP60 was increased in the peri-implantitis group HSP32 immunolabelling slightly decreased in peri-implant and peri-implantitis gingiva. Labelling for HSP72 was undetectable in all three groups. In conclusion, we observed in peri-implantitis a clearly enhanced immunolabelling of two specific HSPs, HSP25 and HSP60, restricted to gingival epithelium and this could indicate a signal of local altered homeostasis.

Stress proteins such as HSP70 members (HSP72 and GRP75) and metallothionein (MT) protect the kidney against oxidative damage and harmful metals, whereas inducible nitric oxide synthase (iNOS) regulates tubular functions. A single dose of mercuric chloride (HgCl2) can cause acute renal failure in rats, its main target being the proximal tubule. Oxidative damage has been proposed as one of its pathogenic mechanisms. In this study we tested whether melatonin (MEL), a powerful antioxidant compound, is effective against HgCl2 nephrotoxicity. Rats were treated with saline, HgCl2 (3.5 mg/kg), MEL (5 mg/kg), and MEL + HgCl2 and examined after 24 hr for HSP72, GRP75, MT, and iNOS by immunohistochemistry and immunoblotting. Tubular effects of the treatment were then characterized by ultrastructure. In the HgCl2 group, all markers were overexpressed in convoluted proximal tubules and sometimes in distal tubules. In the MEL + HgCl2 group, GRP75 and iNOS decreased in convoluted and straight proximal tubules, whereas HSP72 and MT persisted more than the saline and MEL-only groups. Tubular damage and mitochondrial morphometry were improved by MEL pretreatment. In conclusion, the beneficial effect of MEL against HgCl2 nephrotoxicity was outlined morphologically and by the reduction of the tubular expression of stress proteins and iNOS. These markers could represent sensitive recovery index against mercury damage. (J Histochem Cytochem 54:1149-1157, 2006)

The aim of this study was to evaluate the adverse effects of cyclosporine A (CsA) toward renal morphogenesis and to test the renoprotective natural antioxidants such as provinol (PV). Pregnant rats were divided into four groups. Group I was injected SC with olive oil. Group II was treated with oral administration of PV and was used as control. Group III animals were injected SC daily with CsA, and group IV animals were injected daily with CsA and PV for 21 days of pregnancy. Five pups per litter were killed and the kidneys removed and treated by morphological and immunohistochemical (IHC) methods. IHC analysis considered two proteins responsible for nephrotoxicity in adult rats: inducible nitric oxide (iNOS) and matrix metalloproteinase-2 (MMP2). Pregnancy outcomes among CsA-treated rats demonstrated a reduced number of pups. Pups that were exposed antenatally to CsA presented several pathologic findings in all immature parenchyma and an increase in iNOS and MMP2 expression. These side effects were not observed in kidney of litters born from CsA + PV-treated mothers. Our study indicates that CsA induces morphological alterations in renal parenchyma of neonates and that PV plays a protective role against these side effects.

Context and Objective: It has been previously demonstrated that aldosterone may possess a strong profibrotic action in vitro and in animal models of genetic or experimental hypertension. Our aim was to evaluate whether such a profibrotic action is present also in the human microcirculation. Design and Patients: We investigated 13 patients with primary aldosteronism, seven patients with essential hypertension, and 10 normotensive controls. All subjects were submitted to a biopsy of gluteal sc fat tissue. Small resistance arteries were dissected and mounted on an isometric myograph, and the tunica media to internal lumen ratio was measured. Main Outcome Measures: The total collagen content within the tunica media was detected (Sirius red staining and image analysis), and collagen subtypes were evaluated using polarized light microscopy; under this condition thicker type I collagen fibers appear orange or red, whereas thinner type III collagen fibers are yellow or green. Results: Tunica media to internal lumen ratio was significantly increased in primary aldosteronism and in essential hypertension compared with normotensive controls. Clinic blood pressure values were similar in primary aldosteronism and in essential hypertension, and greater than in normotensive controls. Normotensive controls had less total and type III collagen (3.23 0.58 and 1.60 0.22%, respectively) in respect to the two hypertensive groups (P 0.001). Total collagen and type III vascular collagen were significantly greater in primary aldosteronism (total collagen, 8.17 1.38%; type III collagen, 6.06 0.74%; P 0.05) than in essential hypertension (total collagen, 6.84 1.15%; type III collagen, 5.25 0.80%). Conclusions: Our results indicate that, in small resistance arteries of patients with primary aldosteronism, a pronounced fibrosis may be detected, even more evident than in blood-pressure-matched patients with essential hypertension. (J Clin Endocrinol Metab 91: 2638–2642, 2006)

Bimoclomol (BIM), is a stress proteins coinducer, that acts synergistically with a mild stressor to activate cytoprotective stress proteins. BIM has been successfully utilized in animal models for the treatment of various nervous, cardiac and cerebrovascular diseases. Mercuric chloride (HgCl2) induces acute renal failure in rats by a single dosage. The present in vivo study was conducted to assess the efficacy of BIM against acute HgCl2 nephrotoxicity. At different times after BIM and/or HgCl2 exposure we evaluated renal morphology and the localization/abundance of three stress proteins (HSP72, GRP75, HSP60) by electron microscopy, immunohistochemistry and Western blot analysis. BIM delivery to rats 6 h before mercury, ameliorated damage to renal ultrastructure, with recovery of tubular and mitochondrial membranes 24 h after mercury treatment. In rats pretreated with BIM prior to HgCl2 exposure, HSP72 was significantly overexpressed in proximal tubules in a time-dependent manner. In contrast, the amounts of GRP75 and HSP60 after BIM pretreatment were comparable to the group treated with mercury alone, but these stress proteins had translocated to the nuclei at 14 and 24 h, respectively. These novel findings suggest that BIM mitigates HgCl2 nephrotoxicity in rats through the early recruitment of stress proteins in midcortical proximal tubules that are the main renal mercury-targets.