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In the kidney, aging is characterized by the development of structural changes, including glomerulosclerosis and interstitial fibrosis. Transforming growth factor-beta1 (TGF-beta1) is known to play a critical role in the genesis of these alterations in pathologic conditions. The present experiments were designed to test the hypothesis that TGF-beta1 may be involved in the development of age-related histopathologic changes in rat kidney, and that captopril, an angiotensin-converting enzyme inhibitor, may influence the progression of glomerular and interstitial lesions. In this study, 3-, 18-, 24-, and 30-mo-old rats were examined, and an age-related increase in urinary protein excretion was found; plasma creatinine and systolic BP did not change. Significant structural changes, including glomerular sclerosis and interstitial fibrosis, were found in the group of aged rats (24- and 30-mo-old). Immunostaining for TGF-beta in the renal cortex interstitium was increased in the group of 24-mo-old rats, with a parallel increase in TGF-beta1 mRNA expression, measured with reverse-transcription PCR. Captopril-treated animals showed a statistically significant decrease in urinary protein excretion but no significant changes in BP. Moreover, captopril reduced the extent of interstitial fibrosis, but did not affect the degree of glomerulosclerosis. A significant inhibition of TGF-beta1 mRNA expression was observed in the captopril-treated animals. These findings suggest that TGF-beta1 may act as a fibrogenic growth factor that could be responsible, at least partially, for the renal interstitial fibrosis associated with aging. Treatment with captopril might delay the progression of these lesions.

We selected a Leishmania tropica cell line resistant to daunomycin (DNM) that presents a multidrug-resistant (MDR) phenotype characterized by overexpression of a P-glycoprotein of 150 kDa. The resistant line overexpressed an MDR-like gene, called ltrmdr1 , located in an extrachromosomal circular DNA. DNM uptake experiments using laser flow cytometry showed a significant reduction in drug accumulation in the resistant parasites. The initial stages of the interaction of DNM with membranes from wild-type and DNM-resistant parasites were defined by a rapid kinetic stopped-flow procedure which can be described by two kinetic components. On the basis of a previous similar kinetic study with tumor cells, we ascribed the fast component to rapid interaction of DNM with membrane surface components and the slow component to passive diffusion of the drug across the membranes. The results reported here indicate that entrance of DNM into wild-type parasites was facilitated in respect to the resistant ones. We propose that resistance to DNM in L. tropica is a multifactorial event involving at least two complementary mechanisms: an altered drug membrane permeability and the overexpression of a protein related to P-glycoprotein that regulates drug efflux.

Objective. This study was designed to explore relationships of resonance frequency analysis (RFA)—assessed implant stability (ISQ values) with bone morphometric parameters and bone quality in an ex vivo model of dental implants placed in human femoral heads and to evaluate the usefulness of this model for dental implant studies. Material and Methods. This ex vivo study included femoral heads from 17 patients undergoing surgery for femoral neck fracture due to osteoporosis (OP) ( ) or for total prosthesis joint replacement due to severe hip osteoarthrosis (OA) ( ). Sixty mm Dentsply Astra implants were placed, followed by RFA. CD44 immunohistochemical analysis for osteocytes was also carried out. Results. As expected, the analysis yielded significant effects of femoral head type (OA versus OA) ( ), but not of the implants ( ) or of the interaction of the two factors ( ). Bonferroni post hoc comparisons showed a lower mean ISQ for implants in decalcified ( ) heads than in fresh ( ) or fixated ( ) heads (both ). The ISQ score (fresh) was significantly higher for those in OA ( ) versus OP ( ) heads. However, mixed linear analysis showed no significant association between ISQ scores and morphologic or histomorphometric results ( in all cases), and no significant differences in ISQ values were found as a function of the length or area of the cortical layer (both ). Conclusion. Although RFA-determined ISQ values are not correlated with morphometric parameters, they can discriminate bone quality (OP versus OA). This ex vivo model is useful for dental implant studies., This investigation was partially supported by Research Group no. CTS-138 (Junta de Andalucía, Spain).

[Objectives] The aim of this pilot study was to investigate associations between IL-6 and COX-2 expression in gingival biopsies and both clinical diagnosis and genotypes in the IL-6 and COX-2 genes. [Design] A case-control study included 41 gingival biopsies obtained from Caucasian patients grouped according to clinical diagnosis of gingival health (n = 10), gingivitis (n = 15) or chronic periodontitis (n = 16). Immunohistochemistry analyses were performed to determine COX-2 expression in lamina propria, IL-6 expression in lamina propria and gingival epithelium and level of inflammatory cell infiltrate. Individual DNA was extracted and genotyped by real-time PCR for IL6 SNPs rs 2069827 and rs 2069825 and for COX-2 rs 6681231. [Results] The percentage of cellular COX-2 expression was associated with the extent of periodontal disease (Arbes index p = 0.026) and inflammatory infiltrate (p, This study was partially undertaken at the UCL Eastman Dental Institute, which received a proportion of funding from the Department of Health’s National Institute of Health Research (NIHR) Biomedical Research Centres funding scheme.

Renal fibrosis is characterized by an increased number of fibroblasts and excessive deposition of extracellular matrix. Apoptotic cell death is a physiological mechanism to limit cell numbers, and an insufficient rate of death may contribute to fibroblast accumulation. However, little is known about the regulation of renal fibroblast survival. The authors have studied the interaction of cytokines and the Fas receptor in the regulation of apoptosis of renal fibroblasts and have observed that murine renal fibroblasts express Fas and the Fas ligand. Tumor necrosis factor alpha (TNFalpha) and agonistic anti-Fas antibodies induce apoptosis of renal fibroblasts in a time- and dose-dependent manner. Serum contains survival factors for renal fibroblasts. Both serum deprivation and TNFalpha increase the sensitivity to Fas-induced death and the expression of fas mRNA and Fas receptor. By contrast, insulin-like growth factor-1 decreases apoptosis induced by both serum deprivation and Fas activation and partially prevents the increase in Fas receptor expression induced by serum deprivation. Murine renal fibroblasts express constitutively both fas ligand mRNA and cell-surface Fas ligand, but the authors could not demonstrate a role for Fas ligand in the autocrine regulation of fibroblast survival. These data suggest that Fas and other cytokines cooperate to regulate renal fibroblast apoptosis. Modulation of the Fas death-signaling pathway in renal fibroblasts could represent a new therapeutic target for renal fibrosis.

The present experiments were devoted to analyzing the mechanisms involved in the captopril-dependent inhibition of human mesangial cell proliferation. Studies were performed in cultured human mesangial cells incubated with captopril, an angiotensin II-converting enzyme inhibitor with antioxidant properties, lisinopril, a non-antioxidant angiotensin II-converting enzyme inhibitor, and tocopherol, a pure antioxidant. Both angiotensin II-converting enzyme inhibitors significantly inhibited fetal calf serum-induced [ 3 H ]thymidine uptake by human mesangial cells, in a dose- and time-dependent manner, an effect which was not observed with tocopherol. The antiproliferative effect of captopril and its ability to block tyrosine phosphorylation of human mesangial cells proteins were significantly greater than those of lisinopril. Moreover, captopril significantly prevented the fetal calf serum-induced tyrosine phosphorylation of pp60 c- src . The present results suggest that the antiproliferative ability of captopril does not completely depend on its angiotensin II-converting enzyme inhibitor properties, pointing to a possible interaction of the drug with the intracellular mechanisms responsible for the transmission of the proliferative signals.

The enzyme poly(ADP-ribose) polymerase (PARP-1) participates in the repair of DNA damaged by genotoxic agents such as oxygen-derived free radicals. If the allograft suffers pretransplant cold ischemia and subsequent ischemia-reperfusion injury (IR), overactivation of PARP-1 can be induced, which may lead to an increase in acute tubular necrosis (ATN) and a delay in total recovery of renal function (RRF) of the transplanted organ. We studied the nuclear expression of PARP-1 in tubular cells by immunohistochemistry with the monoclonal antibody PAR01 in 104 kidney transplant biopsies from allografts with ATN. In 50% of biopsies with ATN,50% of tubular nuclei were PARP-1+; only 9.6% of biopsies were negative. The increase in the immunohistochemical expression of PARP-1 showed a statistically significant relationship with the duration of cold ischemia, with serum creatinine levels, and with the time required to achieve effective diuresis (P.0001, Spearman test). Cold ischemia of24 hours and serum creatinine levels1.7 mg/dL showed a statistically significant relationship with the highest PARP-1 expression levels (2.83 +/- 0.4 vs 1.36 +/- 0.8, P.0001, Mann-Whitney U test). We conclude that PARP-1 plays an important role in ATN and RRF and is related to the extent and severity of ATN and to the renal allograft function.

We report the case of a 74-year-old white male patient who had worn an overdenture for the previous 6 years, retained by 4 screwed implants and a bar, who presented with an exophytic multilobed lesion of 2.5 × 2.0 cm on the anterior aspect of 1 implant neck, which was surrounded by pink-reddish tissue. All of the soft tissue around the implant was removed until the periosteum was reached. Histologic examination of the lamina propria revealed a cellular proliferation with imprecise boundaries, dense stromal component composed of spindle- to round-shaped mononucleated cells (fibroblasts and monocytes/macrophages), abundant multinucleated giant cells surrounding microscopic hemorrhagic foci, and deposits of hemosiderin; the diagnosis was peripheral giant-cell reparative granuloma (PGCG). Giant cells share the immunohistochemical expression of monocyte/macrophage markers (CD68, calprotectin [Mc387]) and osteoclastic cell markers (tartrate-resistant acid phosphatase, cathepsin K, and microphthalmia-associated transcription factor). After 6 months of follow-up, no bone resorption or recurrence of implant loss was observed. There have been only 12 case reports on dental implant–associated PGCG. Research results to date indicate that there may be little difference in immunophenotype among the giant cells of PGCG, central giant cell reparative granuloma, and peri-implant osteolysis. In conclusion, the immunohistochemical study confirms an osteoclast like giant cells phenotype differentiation in PGCG.

We test the hypothesis that PARP inhibition can decrease acute tubular necrosis (ATN) and other renal lesions related to prolonged cold ischemia/reperfusion (IR) in kidneys preserved at 4°C in University of Wisconsin (UW) solution. Material and Methods. We used 30 male Parp1+/+ wild-type and 15 male Parp10/0 knockout C57BL/6 mice. Fifteen of these wild-type mice were pretreated with 3,4-dihydro-5-[4-(1-piperidinyl)butoxyl]-1(2H)-isoquinolinone (DPQ) at a concentration of 15 mg/kg body weight, used as PARP inhibitor. Subgroups of mice were established (A: IR 45 min/6 h; B: IR + 48 h in UW solution; and C: IR + 48 h in UW solution plus DPQ). We processed samples for morphological, immunohistochemical, ultrastructural, and western-blotting studies. Results. Prolonged cold ischemia time in UW solution increased PARP-1 expression and kidney injury. Preconditioning with PARP inhibitor DPQ plus DPQ supplementation in UW solution decreased PARP-1 nuclear expression in renal tubules and renal damage. Parp10/0 knockout mice were more resistant to IR-induced renal lesion. In conclusion, PARP inhibition attenuates ATN and other IR-related renal lesions in mouse kidneys under prolonged cold storage in UW solution. If confirmed, these data suggest that pharmacological manipulation of PARP activity may have salutary effects in cold-stored organs at transplantation., Funding: This research was supported by CTS no. 138 Research Group and from the Carlos III Health Institute of the Spanish Ministery of Health and Consumer Affairs (Red de Investigación Renal, REDinREN 012/0021/0025). “FEDER una manera de hacer Europa”.

The purpose of this study is to evaluate the vertical and horizontal graft bone resorption (GR) in grafted maxillary sinuses and the marginal bone loss (MBL) around implants placed in the sinuses with different prosthetic connections and to determine the effect of other clinical factors on these tissue responses at 6 and 18 months postloading.A total of 254 implants were placed in 150 grafted maxillary sinuses of 101 patients (51.5% female) with mean age of 52.2 years (range, 32-82 years). GR and MBL measurements were made in implants placed with two different prosthetic connections (internal and external) at 6 and 18 months postloading. The complex samples general linear model was used to analyze the influence of patient age, gender, smoking habit, history of periodontal disease, implantation timing (simultaneous vs deferred), and prosthetic abutment length on radiographic GR and MBL values.At 18 months postloading, the MBL ranged from 0 mm to 5.89 mm; less than 1 mm was lost around 49.0% (mesial) and 44.3% (distal) of the implants, while no bone was lost around 32.9% (mesial) and 26.7% (distal). The GR was significantly affected by smoking, remnant alveolar bone height, graft length, graft height, gender, and age, and it significantly decreased over time. The MBL was influenced by the type of connection, implantation timing, and prosthetic abutment length. The MBL was greater with longer postloading interval and higher patient age and in smokers.Resorption of grafts that combine autogenous cortical bone with anorganic bovine bone is dependent on the anatomic features of the sinus and is not affected by the time elapsed after the first 6 months. The MBL in implants placed in these grafted areas is time dependent and mainly related to potentially modifiable clinical decisions and patient habits.

RATIONALE:: Proliferation and migration of smooth muscle cells (SMCs) are key steps for the progression of atherosclerosis and restenosis. Cortistatin is a multifunctional neuropeptide belonging to the somatostatin family that exerts unique functions in the nervous and immune systems. Cortistatin is elevated in plasma of patients experiencing coronary heart disease and attenuates vascular calcification. OBJECTIVE:: To investigate the occurrence of vascular cortistatin and its effects on the proliferation and migration of SMCs in vitro and in vivo and to delimitate the receptors and signal transduction pathways governing its actions. METHODS AND RESULTS:: SMCs from mouse carotid and human aortic arteries and from human atherosclerotic plaques highly expressed cortistatin. Cortistatin expression positively correlated with the progression of arterial intima hyperplasia. Cortistatin inhibited platelet-derived growth factor-stimulated proliferation of human aortic SMCs via binding to somatostatin receptors (sst2 and sst5) and ghrelin receptor, induction of cAMP and p38-mitogen-activated protein kinase, and inhibition of Akt activity. Moreover, cortistatin impaired lamellipodia formation and migration of human aortic SMCs toward platelet-derived growth factor by inhibiting, in a ghrelin-receptor- dependent manner, Rac1 activation and cytosolic calcium increases. These effects on SMC proliferation and migration correlated with an inhibitory action of cortistatin on the neointimal formation in 2 models of carotid arterial ligation. Endogenous cortistatin seems to play a critical role in regulating SMC function because cortistatin-deficient mice developed higher neointimal hyperplasic lesions than wild-type mice. CONCLUSIONS:: Cortistatin emerges as a natural endogenous regulator of SMCs under pathological conditions and an attractive candidate for the pharmacological management of vascular diseases that course with neointimal lesion formation. © 2013 American Heart Association, Inc.

Cortistatin is a cyclic-neuropeptide produced by brain cortex and immune cells that shows potent anti-inflammatory activity. In this article, we investigated the effect of cortistatin in two models of experimental autoimmune encephalomyelitis (EAE) that mirror chronic and relapsing-remitting multiple sclerosis. A short-term systemic treatment with cortistatin reduced clinical severity and incidence of EAE, the appearance of inflammatory infiltrates in spinal cord, and the subsequent demyelination and axonal damage. This effect was associated with a reduction of the two deleterious components of the disease, namely, the autoimmune and inflammatory response. Cortistatin decreased the presence/activation of encephalitogenic Th1 and Th17 cells in periphery and nervous system, and downregulated various inflammatory mediators, whereas it increased the number of regulatory T cells with suppressive effects on the encephalitogenic response. Moreover, cortistatin regulated glial activity and favored an active program of neuroprotection/regeneration. We further used cortistatin-deficient mice to investigate the role of endogenous cortistatin in the control of immune responses. Surprisingly, cortistatin-deficient mice were partially resistant to EAE and other inflammatory disorders, despite showing competent inflammatory/autoreactive responses. This unexpected phenotype was associated with elevated circulating glucocorticoids and an anxiety-like behavior. Our findings provide a powerful rationale for the assessment of the efficacy of cortistatin as a novel multimodal therapeutic approach to treat multiple sclerosis and identify cortistatin as a key endogenous component of neuroimmune system. Copyright © 2013 by The American Association of Immunologists, Inc.

Objective To investigate the effect of adipose-derived mesenchymal stromal cells (ASCs) on the activation state of macrophages (MF) in vitro, and the potential therapeutic effect of these cells in experimental colitis and sepsis. Design Murine bone marrow-derived macrophages were cultured with ASCs or with ASC conditioned media (ASC-MF) and characterised for the expression of several regulatory macrophage markers, including enzymes and cytokines, and for their immunomodulatory capacity in vitro. The therapeutic effect was investigated of ASC-MF in two models of experimental inflammatory colitis induced by trinitrobenzene sulphonic acid and dextran sodium sulphate, and in polymicrobial sepsis induced by caecal ligation and puncture. Results ASC-MF showed a phenotype that clearly differed from the classically activated macrophages or the alternatively activated macrophages induced by interleukin (IL)-4, characterised by high arginase activity, increased production of IL-10 upon restimulation and potent immunosuppressive activity on T cells and macrophages. Activation of cyclo-oxygenase-2 on ASCs seems to be critically involved in inducing this phenotype. Systemic infusion of ASC-MF inhibited colitis in mice, reducing mortality and weight loss while lowering the colonic and systemic levels of inflammatory cytokines. Importantly, therapeutic injection of ASC-MF in established chronic colitis alleviated its progression and avoided disease recurrence. Moreover, ASC-MF protected from severe sepsis by reducing the infiltration of inflammatory cells into various organs and by downregulating the production of several inflammatory mediators, where ASC-MF-derived IL-10 played a critical role. Conclusion ASCs induce a distinct regulatory activation state of macrophages which possess potent immunomodulatory ability and therapeutic potential in inflammatory bowel diseases and sepsis.

Immunohistochemical techniques were used to study the presence of ciclosporin A (CsA) and leukocyte subsets in 36 posttransplant renal biopsy specimens histologically diagnosed as acute graft rejection. Glomeruli from patients with CsA deposits contained more leukocytes (p < 0.05) than glomeruli from tissues without deposits. In contrast, the interstitium from patients without deposits contained significantly more B lymphocytes than interstitia from kidneys with CsA deposits. In both glomeruli and interstitia, the CD4/CD8 ratios were higher in tissues without deposits, although the difference was not significant. The plasma levels of creatinine increased with the intensity of renal CsA deposits, and significantly more patients on hemodialysis had deposits as compared with patients not on hemodialysis. Our findings suggest two types of acute nonvascular rejection: (1) predominantly interstitial, with a good prognosis, characterized by low numbers of intrarenal CsA deposits and a predominance of B lymphocytes and (2) predominantly glomerular, with a poor prognosis, characterized by high levels of intrarenal CsA and a predominance of CD8-positive cells and macrophages.

The aim of this study is to determine whether periodontal clinical parameters, periodontal bacterial profiles, and inflammatory infiltrate in placental chorionic villi are associated with adverse pregnancy results.The authors designed an observational case-control study in 244 postpartum females: mothers with preterm/low-birth weight newborns (n = 91 cases) and mothers with full-term, normal-weight infants (n = 153 controls). Sociodemographic, gynecologic, and periodontal variables were gathered for all participants. Data on placental inflammatory infiltrate in biopsies from 68 cases and 65 controls and the gingival bacterial profile in mothers with periodontitis were gathered, detecting associations with bivariate analyses and constructing a multiple logistic regression model with the number of positive inflammatory cells as the dependent variable.Periodontal values were significantly worse in cases versus controls. Numbers of leukocyte subsets per square millimeters in maternal and fetal vascular spaces were similar between cases and controls. CD45 in maternal placental space was related to the presence of periodontitis (P = 0.029) but not to case or control group (P = 0.264). The anaerobic and commensal bacterial profile in mothers with periodontitis was similar between the groups.Periodontal disease was more severe and a periodontitis diagnosis more frequent in mothers with preterm or low-birth weight versus normal delivery. No differences in anaerobic or commensal bacterial profile were found between mothers with periodontitis in the two groups. Local placental factors, such as the nature of the inflammatory infiltrate and slightly higher expression of cyclooxygenase-2 in the females with these adverse pregnancy outcomes, may be related to a subclinical proinflammatory status that could contribute to triggering premature labor.

Ghrelin is an important gastrointestinal hormone that regulates feeding and metabolism. Moreover, ghrelin is produced by immune cells and shows potent anti-inflammatory activities. Here, we investigated its effect in two models of experimental autoimmune encephalomyelitis (EAE) that mirror chronic and relapsing-remitting multiple sclerosis. A short systemic treatment with ghrelin after the disease onset reduced clinical severity and incidence of both forms of EAE, which was associated with a decrease in inflammatory infiltrates in spinal cord and in the subsequent demyelination. This therapeutic effect was exerted through the reduction of the autoimmune and inflammatory components of the disease. Ghrelin decreased the presence/activation of encephalitogenic Th1 and Th17 cells in periphery and nervous system, down-regulated various inflammatory mediators, and induced regulatory T cells. In summary, our findings provide a powerful rationale for the assessment of the efficacy of ghrelin as a novel therapeutic approach for treating multiple sclerosis through distinct immunomodulatory mechanisms and further support the concept that the neuroendocrine and immune systems crosstalk to finely tune the final immune response of our body.

The purpose was to analyse the cardiac and renal capillary density and glomerular morphology resulting from a chronic excess or deficiency of thyroid hormones (THs) in rats. We performed histopathological, morphometrical and immunohistochemical analyses in hypothyroid and hyperthyroid rats to evaluate the density of mesenteric, renal and cardiac vessels at 4 weeks after induction of thyroid disorders. The main angiogenic factors in plasma, heart and kidney were measured as possible mediators of vascular changes. Mesenteric vessel branching was augmented and decreased in hyper- and hypothyroid rats respectively. The numerical density of CD31-positive capillaries was higher in left and right ventricles and in cortical and medullary kidney from both hyper- and hypothyroid rats vs controls. Numbers of podocytes and glomeruli per square millimetre were similar among groups. Glomerular area and percentage mesangium were greater in the hyperthyroid vs control or hypothyroid groups. No morphological renal lesions were observed in any group. Vascularisation of the mesenteric bed is related to TH levels, but an increased capillarity was observed in heart and kidney in both thyroid disorders. This increase may be produced by higher tissue levels of angiogenic factors in hypothyroid rats, whereas haemodynamic factors would predominate in hyperthyroid rats. Our results also indicate that the renal dysfunctions of thyroid disorders are not related to cortical or medullary microvascular rarefaction and that the proteinuria of hyperthyroidism is not secondary to a podocyte deficit. Finally, TH or its analogues may be useful to increase capillarity in renal diseases associated with microvascular rarefaction.

Objective To investigate the role of the antiinflammatory neuropeptide cortistatin in chronic pain evoked by joint inflammation. Methods Thermal and mechanical hyperalgesia was evoked in mouse knee joints by intraplantar injection of tumor necrosis factor α and intraarticular infusion of Freund's complete adjuvant, and the analgesic effects of cortistatin, administered centrally, peripherally, and systemically, were assessed. In addition, the effects of cortistatin on the production of nociceptive peptides and the activation of pain signaling were assayed in dorsal root ganglion cultures and in inflammatory pain models. The role of endogenous cortistatin in pain sensitization and perpetuation of chronic inflammatory states was evaluated in cortistatin-deficient mice. Finally, the effect of noxious/inflammatory stimuli in the production of cortistatin by the peripheral nociceptive system was assayed in vitro and in vivo. Results Expression of cortistatin was observed in peptidergic nociceptors of the peripheral nociceptive system, and endogenous cortistatin was found to participate in the tuning of pain sensitization, especially in pathologic inflammatory conditions. Results showed that cortistatin acted both peripherally and centrally to reduce the tactile allodynia and heat hyperalgesia evoked by arthritis and peripheral tissue inflammation in mice, via mechanisms that were independent of its antiinflammatory action. These mechanisms involved direct action on nociceptive neurons and regulation of central sensitization. The analgesic effects of cortistatin in murine arthritic pain were linked to binding of the neuropeptide to somatostatin and ghrelin receptors, activation of the G protein subunit Gαi, impairment of ERK signaling, and decreased production of calcitonin gene-related peptide in primary nociceptors. Conclusion These findings indicate that cortistatin is an antiinflammatory factor with potent analgesic effects that may offer a new approach to pain therapy in pathologic inflammatory states, including osteoarthritis and rheumatoid arthritis. Copyright © 2013 by the American College of Rheumatology.

Different biomaterials have been suggested for guided bone regeneration (GBR). These might show the ideal properties to let a new bone formation in the grafted area. Among these ideal features, it is essential their controlled resorption in order to be replaced for new vital bone. Bovine bone has been used widely as a good biomaterial for GBR, however there is still an interesting controversy about its resorbable capacity. In this sense, the objective of this study was to examine the behavior of anorganic bovine bone (ABB) in long-term maxillary sinus graft healing and study its relationship with morphological and morphometrical variables.Seventeen maxillary sinus augmentation procedures were performed in patients. Bone cores were obtained from implant receptor sites at 6 months, 3 years, and 7 years of implant placement for histological, morphometric, and immunohistochemical (tartrate resistant acid phosphatase [TRAP]/cathepsin K/CD68) studies.The percentages of bone, ABB particles, connective tissue, osteocytes, and osteoblasts in maxillary sinus grafts were similar at 6 months, 3 years, and 7 years. A progressive and significant decrease was detected in osteoclasts (p = .05, Kruskal-Wallis test), TRAP and cathepsin K expression (p = .014 and p = .021, respectively), and osteoid lines (p = .038).According to these data, a decrease in osteoclasts over time may, partially, explain the ABB persistence observed in core biopsies. Further studies with more cases and different graft maturation times are required to elucidate the resorption rates and cell events underlying these phenomena.

The objective of this study is to compare cyclooxygenase-2 (COX-2) protein expression in gingival biopsies from patients with chronic periodontitis (CP), patients with gingivitis (GV), and individuals with no periodontal disease (control group) and to establish its relationship with clinical variables and connective tissue loss in the lamina propria.A cross-sectional and analytic study was conducted in 108 gingival biopsies from 52 patients with CP, 39 with GV, and 17 controls. All biopsies were processed for conventional histopathologic study, immunohistochemical determination of COX-2 protein expression, and automatic quantification of connective tissue by image analysis.The protein expression of COX-2, mainly produced by plasma cells and monocytes, was significantly related to the presence of periodontal disease, bleeding index, intensity of inflammatory infiltrate, and loss of connective tissue in the lamina propria of gingival biopsies (P0.01, Spearman test). COX-2 expression was also directly correlated with attachment loss (P0.05, Spearman test).COX-2 protein expression is higher in patients with GV and CP than in individuals without periodontal disease and is inversely correlated with the amount of connective tissue in the lamina propria as determined by image analysis. This finding suggests that COX-2 participates in mechanisms and pathway signaling related to the destruction of fibrillar support structures of the periodontium.

Immunohistochemical techniques were used to study the presence of cyclosporin A (CsA) and leukocyte subsets in 30 gingival biopsies of renal transplant subjects with gingival overgrowth (GO). Statistical analysis revealed significant differences in the total number of inflammatory cells determined by monoclonal antibody CD45, the monocyte/macrophage (CD68) subset, the plasmatic cells (EMA), and the total of T-lymphocytes (CD3) (P0.001, Student t test) between the treated subjects and the healthy control group. Differences were found in the helper/inducer T lymphocytes CD4 (P0.001 Student t test) and cytotoxic/suppressor T lymphocyte (CD8) (P0.01, Student t test) subsets between both groups. The CD4/CD8 ratio was greater in the transplant subjects than in the control group (1.82 +/- 0.16 versus 1.35 +/- 0.05 respectively) (P0.05 Student t test). There was no significant difference in the populations CD16+, CD57+, and CD20+. The CD45+ CD4+, and CD68+ cells increased in number along with the degree of GO. The number of epithelial cells/mm2 which displayed a deposit of CsA increased in accordance with the degree of GO (P0.05, Kruskal-Wallis's test). Likewise, the intraepithelial deposit of CsA in the GO region was found to be related to the inflammatory infiltrate CD4+, CD8+, and CD68+ (r = 0.7432; r = 0.7346; r = 0.77005, respectively). Our findings suggest that the intraepithelial deposit of CsA and the inflammatory infiltrate play a predominantly pathogenic role and are both related to the degree of GO.

Summary The influence of immunosuppressant therapy and of the presence of CMV genome on thedistribution of lymphoid subpopulations of the inflammatory infiltrate in renal graft rejection was analyzed, as was the role of both factors in the evolution and survival of the graft. The study included 22 patients treated with Cyclosporin A (CsA) and 22 patients treated with Azatbioprine (AZA). Inflammatory infiltrate was studied by immunostaining with a panel of monoclonal antibodies, and CMV DNA was detected by in situ hybridization on tissue sections. In patients treated with CsA, increased cellularity was found at both glomerular and interstitial levels, consisting mainly of macrophages and T-cells, which was consistent with the higher rate of glomerulointer stitial rejection found in this group. In contrast, the vascular type of rejection predominated in AZA treated patients. However, the presence of CMV DNA did not influence the phenotype of the inflammatory infiltrate, and was not associated with any specific lesion. Furthermore, the final outcome of the renal graft was independent of the detection of CMV. Therefore, this study provides no evidence of any active role of the CMV genome in renal graft rejection, and suggests that therapy should be adapted to the type of rejection as defined on morphologic and immunopbenotypic grounds.

The synthesis of a series of pyrrole and imidazole retinoids is reported. The compounds were obtained by means of reactions of phosphoranes or phosphonates with aldehydes. Several of these retinoids were evaluated against different tumor systems and some were found to have an activity comparable to that of all-trans retinoic acid.

We report an unusual case of hemangiopericytoma-like dermatofibroma in the right shoulder of an 82-year-old patient with a well-defined nodular growth located in the dermis. Microscopic study revealed a band of haphazardly arranged cells with a vascular component of gaping, simple, endothelial-lined vascular structures with intervening postcapillary venules and capillary-sized slit-like "staghorn" vascular channels filled with erythrocytes; abundant mast cells were also observed. The neoplasm cells were positive for CD68 and Factor XIIIA and negative for CD34. Few data have been published on the presence of abundant mast cells (tryptase and CD117 positive) in these neoplasm. The differential diagnosis of this entity should consider other spindle cell neoplasm, including hemangiopericytoma/solitary fibrous tumor, dermatofibrosarcoma protuberans, myopericytoma, angioleiomyoma, amelanotic melanoma, pecoma, and benign and malignant peripheral nerve tumors. We present an infrequent case of dermatofibroma with a vascular pattern resembling hemangiopericytoma and the presence of abundant mast cells, which may be responsible for this vascular component.

This study assessed salt sensitivity, analyzing the effects of an increased saline intake on hemodynamic, morphological, and oxidative stress and renal variables in experimental thyroid disorders. Six groups of male Wistar rats were used: control, hypothyroid, hyperthyroid, and the same groups treated with salt (8% via food intake). Body weight, blood pressure (BP), and heart rate (HR) were recorded weekly for 6 wk. Finally, BP and HR were recorded directly, and morphological, metabolic, plasma, and renal variables were measured. High-salt intake increased BP in thyroxine-treated rats but not in control or hypothyroid rats. High-salt intake increased cardiac mass in all groups, with a greater increase in hyperthyroid rats. Urinary isoprostanes and H2O2were higher in hyperthyroid rats and were augmented by high-salt intake in all groups, especially in hyperthyroid rats. High-salt intake reduced plasma thyroid hormone levels in hyperthyroid rats. Proteinuria was increased in hyperthyroid rats and aggravated by high-salt intake. Urinary levels of aminopeptidases (glutamyl-, alanyl-, aspartyl-, and cystinylaminopeptidase) were increased in hyperthyroid rats. All aminopeptidases were increased by salt intake in hyperthyroid rats but not in hypothyroid rats. In summary, hyperthyroid rats have enhanced salt sensitivity, and high-salt intake produces increased BP, cardiac hypertrophy, oxidative stress, and signs of renal injury. In contrast, hypothyroid rats are resistant to salt-induced BP elevation and renal injury signs. Urinary aminopeptidases are suitable biomarkers of renal injury.

Smoothelin is a cytoskeletal protein of differentiated smooth muscle cells with contractile capacity, distinguishing it from other smooth muscle proteins, such as smooth muscle actin (SMA).To evaluate the expression of smoothelin and SMA in the skin in order to establish specific localizations of smoothelin in smooth muscle cells with high contractile capacity and in the epithelial component of cutaneous adnexal structures.Immunohistochemical analysis (smoothelin and SMA) was performed in 18 patients with normal skin.SMA was expressed by the vascular structures of superficial, deep, intermediate and adventitial plexuses, whereas smoothelin was specifically expressed in the cytoplasm of smooth muscle cells of the deepest vascular plexus and in no other plexus of the dermis. The hair erector muscle showed intense expression of smoothelin and SMA. Cells with nuclear expression of smoothelin and cytoplasmic expression of SMA were observed in the outer root sheath of the inferior portion of the hair follicles and intense cytoplasmic expression in cells of the dermal sheath to SMA.We report the first study of smoothelin expression in normal skin, which differentiates the superficial vascular plexus from the deep. The deep plexus comprises vessels with high contractile capacity, which is important for understanding dermal hemodynamics in normal skin and pathological processes. We suggest that the function of smoothelin in the outer root sheath may be to enhance the function of SMA, which has been related to mechanical stress. Smoothelin has not been studied in cutaneous pathology; however we believe it may be a marker specific for the diagnosis of leiomyomas and leiomyosarcomas of the skin. Also, smoothelin could differentiate arteriovenous malformations of cavernous hemangioma of the skin.

Background: It has been demonstrated that human umbilical cord stromal stem cells (UCSSCs) are bio-equivalent to bone marrow mesenchymal stem cells. However, little is known about their tissue origin or in vivo functions, and data on their expansion properties are limited due to early senescence in the culture methods described to date. Methods: UC sections and cultured UCSSCs were analyzed with a panel of 12 antibodies. UCSSCs were grown in low-FCS containing medium at 5% or 21% oxygen and were assayed for their clonogenic properties, karyotype stability, expression of specific cellular markers, and multi-lineage potential. UCSSC contractile properties were evaluated by using collagen gel contraction assays under cytokine stimulus. Results: Immunohistochemistry studies showed that the UCSSCs were derived from the Wharton's jelly and not from the vascular smooth muscle sheath of the blood vessels. UCSSC growth properties were increased in a 5% oxygen atmosphere in comparison to normoxic culture conditions. In both culture conditions, UCSSCs were CD14-, CD34-, and CD45-negative while expressing high levels of CD73, CD90 and CD105 and maintaining their differentiation potentialities. UCSSCs expressed alpha smooth muscle actin and behaved as functional myofibroblasts when cellular contraction was challenged with appropriate stimuli. Conclusions: UCSCs are mesenchymal stem cells that reside in the perivascular area of Wharton's jelly and are phenotypically and functionally related to myofibroblasts. © 2010 Elsevier Ltd. All rights reserved., This research project was supported by a grant from the Junta de Andalucía (P06-CTS-1385) and partially supported by research group grant CTS-138 (Junta de Andalucía, Spain)

The objective of this study was to determine, using digital panoramic radiographs, whether the bone level at the alveolar crest is related to the mandibular bone density and/or to vitamin D receptor (VDR) gene polymorphisms. We analyzed 319 digital panoramic radiographs from the same number of patients. Alveolar bone level was expressed as percentage of root length. The mandibular cortical width index was calculated as a measure of mandibular bone density, and, in 72 randomly selected cases, the haplotype of the VDR gene (BsmL) was determined by polymerase chain reaction. Alveolar bone level was not related to the mandibular cortical width index (p = 0.568) or VDR gene expression (p = 0.575). Bone loss was greater in smokers than in non-smokers (p = 0.036), and the mandibular cortical width index was higher in males (p = 0.04), the older age group (p = 0.032), and in those with more teeth (p = 0.01). Multivariate analysis confirmed the association between these variables and alveolar bone loss. Alveolar bone loss showed no significant relationship with the mandibular bone density evaluated on digital panoramic radiographs or with VDR genotype (BsmL) in Caucasian females and males aged under 47 years.

RWPs (red wine polyphenols) exert antihypertensive effects and improve endothelial function by reducing the plasma levels of ET-1 (endothelin-1) and the subsequent vascular production of O(2)(•-) (superoxide anion). Our present study was designed to evaluate whether RWPs act directly in the vascular wall improving endothelial dysfunction and O(2)(•-) production induced by ET-1 and to analyse the compounds responsible for these protective effects. We incubated rat isolated aortic rings in the presence or absence of ET-1 (10 nM) and RWPs (10(-4) to 10(-2) g/l) or catechin (0.2 μM), epicatechin (10 μM) and resveratrol (0.1 μM). ET-1 reduced the relaxant responses to acetylcholine, increased intracellular O(2)(•-) production, NADPH oxidase activity and protein expression of NADPH oxidase subunit p47phox. All these changes were prevented by RWPs. The preventive effects of RWPs were unaffected by co-incubation with either ICI-182780, an ER (oestrogen receptor) antagonist, or GW9662, a PPARγ (peroxisome-proliferator-activated receptor γ) antagonist. RWPs inhibited the phosphorylation of the mitogen-activated protein kinase, ERK1/2 (extracellular signal-regulated kinase 1/2), a key regulator of p47phox expression in response to ET-1. When the isolated polyphenols were tested, at the concentrations found in 10(-2) g/l RWPs, only epicatechin prevented endothelial dysfunction and all biochemical changes induced by ET-1 in the vascular wall. Taken together, these results indicate that RWPs prevent ET-1-induced vascular O(2)(•-) production by reducing overexpression of p47phox and the subsequent increased NADPH oxidase activity, leading to improvement in endothelial function. The effects of RWPs appear to be independent of ER and PPARγ activation and are related to ERK1/2 inhibition.

The synthesis of a series of pyrrole and imidazole retinoids is reported. The compounds were obtained by means of reactions of phosphoranes or phosphonates with aldehydes. Several of these retinoids were evaluated against different tumor systems and some were found to have an activity comparable to that of all-trans retinoic acid.

BACKGROUND We investigated the contribution of the sympathetic tone to the hypertension induced by chronic administration of buthionine sulfoximine (BSO) and characterized this model in mice. METHODS Three experiments were performed. In experiment I, four groups of CBA-C57 male mice were used: controls and three groups that received oral BSO at 5, 10, or 20 mmol/l. In experiment II, the alpha(1)-adrenergic blocker prazosin was orally administered (10 mg/100 ml) to control and BSO-treated mice. All treatments were maintained for 5 weeks. Body weight (BW), tail blood pressure (BP), and heart rate (HR) were measured weekly. Direct mean arterial pressure (MAP) and morphological, metabolic, plasma, and renal variables were measured at the end of the experiments. In experiment III, the acute response of MAP and HR to the ganglionic blocker pentolinium (10 mg/kg intravenous) was used to further evaluate the sympathetic contribution to BP and HR in control and BSO-treated mice. RESULTS BSO produced dose-related increases in BP (control, 115 +/- 0.5; BSO-5, 141 +/- 0.5; BSO-10, 151 +/- 0.9; BSO-20, 163 +/- 1.1 mm Hg) and HR and augmented plasma noradrenaline, brainstem isoprostane levels, and total urinary isoprostane excretion. BSO did not produce cardiac hypertrophy and did not modify metabolic or plasma variables, or creatinine clearance, proteinuria, or renal morphology. Chronic prazosin markedly reduced MAP (control, 101 +/- 4.7; prazosin, 95 +/- 1.29; BSO-10, 130 +/- 2.9; BSO-10 +/- prazosin, 98 +/- 0.9) and HR. Acute pentolinium produced a greater percentage MAP (control, 43 +/- 4.2; BSO-10, 66 +/- 4.5) and HR decrease in BSO-treated mice vs. controls. CONCLUSION Sympathetic tone plays a major role in the increased BP and HR of BSO hypertensive mice.

Development of new biomaterials is a constant in regenerative medicine. A biomaterial’s surface properties, such as wettability, roughness, surface energy, surface charge, chemical functionalities and composition, are determinants of cell adhesion and subsequent tissue behavior. Thus, the main aim of this study was to analyze the correlation between changes in wettability without topographical variation and the response of osteoblast-like cells. For this purpose oxidized silicon surfaces were methylated to different degrees. Additionally, the influence of nanoroughness, and the subsequent effect of hysteresis on cell behavior, was also analyzed. In this case oxidized silicon pieces were etched with caustic solutions to produce different degrees of nanoroughness. Axisymmetric drop-shape analysis and atomic force microscopy confirmed that the proposed surface treatments increased the nanometer roughness and/or the water contact angles. MG-63 osteoblast-like cells were cultured on the altered surfaces to study proliferation, and for ultrastructural analysis and immunocytochemical characterization. Increasing the nanometer surface roughness or water contact angle enhanced osteoblast behavior in terms of cell morphology, proliferation and immunophenotype, the effect provoked by methylation being more significant than that caused by nanoroughness.

It is difficult to prevent and treat intra- and peri-neural fibrosis after peripheral nerve surgery. Many authors have attempted to develop and verify the effectiveness of substances to decrease the formation of adherences in different tissues.this study aimed to assess the effectiveness of a barrier of oxidized regenerated cellulose (ORC) to reduce adherence and perineural fibrosis in a model of surgical perineural induced fibrosis in rat sciatic nerve in 40 rats. After tissue aggression, the nerve of the right rear limb was wrapped in ORC and the left limb served as control. Animals were killed at 3 and 6 weeks, and nerves and muscle mass were extracted en bloc. Connective tissue was quantified by conventional histopathological techniques and Fibrosis HR(R) automatic image analysis.No significant differences were found in intra- or peri-neural induced fibrosis between control nerves (6.88% and 8.90%, respectively) and treated nerves (6.57% and 9.90%) at 3 or 6 weeks (10.41% and 12.51% in controls; 11.85% and 15.72% in treated nerves). Inflammatory phenomena and granulomatous reactions were more frequent in treated animals.ORC conferred no advantage in prevention of nerve fibrosis and might have interfered with healing.

Grafts with subclinical rejection associated with interstitial fibrosis and tubular atrophy (SCR+IF/TA) show poorer survival than grafts with subclinical rejection without IF/TA (SCR). Aiming to detect differences among SCR+IF/TA and SCR, we immunophenotyped the inflammatory infiltrate (CD45, CD3, CD20, CD68) and used a low-density array to determine levels of T(H)1 (interleukin IL-2, IL-3, gamma-interferon, tumor necrosis factor-alpha, lymphotoxin-alpha, lymphotoxin-beta, granulocyte-macrophage colony-stimulating factor) and T(H)2 (IL-4, IL-5, IL-6, IL-10, and IL-13) transcripts as well as of IL-2R (as marker for T-cell activation) in 31 protocol biopsies of renal allografts. Here we show that grafts with early IF/TA and SCR can be distinguished from grafts with SCR on the basis of the activation of IL-10 gene expression and of an increased infiltration by B-lymphocytes in a cellular context in which the degree of T-cell activation is similar in both groups of biopsies, as demonstrated by equivalent levels of IL-2R mRNA. These results suggest that the up-regulation of the IL-10 gene expression, as well as an increased proportion of B-lymphocytes in the inflammatory infiltrates, might be useful as markers of early chronic lesions in grafts with SCR.

We report the clinical-morphological study of a granular cell tumor in dermal/hypodermal junction and subcutaneous fat left breast of an 83-year-old woman with a family history of breast carcinoma. Mammography study showed a spiculated lesion in the lower inner quadrant with suspicion of malignancy. The results of fine needle puncture-aspiration were inconclusive. Subsequent tumorectomy revealed a poorly-defined indurated lesion of 1.1 × 0.7 cm. The histopathology study showed a proliferation of cells with ample and granular cytoplasm that were positive for S100, CD 68 and inhibin and negative for hormonal receptors. We present a benign lesion that clinically reproduces a breast carcinoma. Keywords Granular cell tumor; Breast; Differential diagnosis; Cutaneous

Myositis ossificans circumscripta is a form of heterotopic ossification that is benign in nature associated to a trauma, but may appear clinically and radiologically as a malignant neoplasm. We describe a rare case of calcifying of myositis ossificans not associated to trauma in a 35-year-old woman with a mass in her upper third and external of right thigh. We discuss some of the difficulties of diagnosis and histological evolution of the lesion. Keywords Myositis ossificans; Thigh; Differential diagnosis; Nontraumatic

Sinus grafting is a technique oriented to facilitate implant placement in posterior atrophic maxillae. Several modifications of the original technique and a wide variety of materials have been proposed; most of them associated with implant survival rates. However, the quality of the bone obtained after the application of certain grafting materials has not been fully elucidated yet. The aims of this multicenter study were to analyse histomorphometrical samples obtained 6 months after sinus grafting using a composite graft consisting of anorganic bovine bone (ABB)+ autologous bone (AB), and to compare these samples with maxillary pristine bone biopsies.Ninety maxillary sinus augmentations were performed for delayed implant placement (N = 90) in 45 consecutive patients (test group). Bone cores were harvested 6 months after grafting for histomorphometric and ultrastructural study. Control pristine bone biopsies were taken from the posterior maxilla of 10 patients (control). Bone radiographic changes were assessed up to 24 months after implant loading.The total mean values after analysis of test cores revealed a proportion of 46.08 + or - 16.6% of vital bone, 42.27 + or - 15.1% of non-mineralized connective tissue, and 37.02 + or - 25.1% of the remaining ABB particles. Significant bone remodeling activities were noticed in sinus grafting samples when compared with pristine bone. A statistically significant difference was observed in the number of osteoid lines between two groups, with higher values in the test one (15.1 + or - 11.48% vs. 2.5 + or - 2.2%, P = 0.0005). Ultrastructural study showed that vital trabecular bone was in intimal contact with ABB particles. Radiographic analysis revealed that the higher the proportion of remaining ABB, the lower the total vertical resorption of the graft.Sinus grafting constitutes an excellent model for the study of de novo bone formation patterns and graft consolidation, when a combination of different bone substitutes is applied. The combination of ABB+AB yields highly satisfactory outcomes from both a clinical and a histologic perspective.

Objective.To assess whether blockade of tumor necrosis factor-α (TNF-α) influences surgical wound healing in a normal mouse experimental model.Methods.Wound healing time course and degree of surgical wound collagenization were measured by morphological techniques and digital image analysis in 80 male SWISS-OF1 mice (40 received subcutaneous etanercept at a dose of 0.1 mg/25g/ at −7, 0, 7, and 14 days).Results.No significant differences were observed between treated and untreated animals in wound healing, re-epithelialization, or formation of inflammatory infiltrate or granulation tissue at days 7, 15, or 20 after surgery. At 20 days, the collagen area was larger in treated versus untreated mice (109029 ± 28489 μm2 vs 79305 ± 19798 μm2, p = 0.026, Mann-Whitney U test).Conclusion.Surgical wounds showed a higher degree of collagenization at 20 days in etanercept-treated versus untreated mice, with no differences in the time course of wound healing. These data suggest that biological therapies to block TNF-α do not affect wound healing and do not need to be suspended during the perioperative period.

Environmental stress conditions are important factors in human health and should be considered in the development of appropriate health policies, since they have been associated with psychological disorders and even with death. A link between stress and changes in 3alpha,5alpha-reduced neurosteroids has been reported. Steroid 5alpha-Reductase (5alpha-R) is the rate-limiting enzyme in the biosynthesis of 3alpha,5alpha-reduced neurosteroids. Using reverse transcription-polymerase chain reaction and immunohistochemistry, 5alpha-R isozymes (5alpha-R1 and 5alpha-R2) mRNA and protein levels were detected in prefrontal cortex of male and female rats after they underwent environmental stresses, i.e., excessive heat, artificial light, and the sensation of immobility in a small space, similar to those found in common workplace situations. Results showed significantly higher 5alpha-R2 mRNA and protein levels in environmentally-stressed versus control rats. Interestingly, a sexual dimorphism in 5alpha-R1 mRNA and protein levels was observed after environmental stress, with an increase in males and a decrease in females. This fact might explain gender differences in the incidence of some type of minor depression.

Cold ischemia time especially impacts on outcomes of expanded-criteria donor (ECD) transplantation. Ischemia-reperfusion (IR) injury produces excessive poly[ADP-Ribose] Polymerase-1 (PARP-1) activation. The present study explored the hypothesis that increased tubular expression of PARP-1 contributes to delayed renal function in suboptimal ECD kidney allografts and in non-ECD allografts that develop posttransplant acute tubular necrosis (ATN). Materials and Methods: Nuclear PARP-1 immunohistochemical expression was studied in 326 paraffin-embedded renal allograft biopsies (193 with different degrees of ATN and 133 controls) and in murine Parp-1 knockout model of IR injury. Results: PARP-1 expression showed a significant relationship with cold ischemia time (r coefficient = 0.603), time to effective diuresis (r = 0.770), serum creatinine levels at biopsy (r = 0.649), and degree of ATN (r = 0.810) (p= 0.001, Pearson test). In the murine IR model, western blot showed an increase in PARP-1 that was blocked by Parp-1 inhibitor. Immunohistochemical study of PARP-1 in kidney allograft biopsies would allow early detection of possible delayed renal function, and the administration of PARP-1 inhibitors may offer a therapeutic option to reduce damage from IR in donor kidneys by preventing or minimizing ATN. In summary, these results suggest a pivotal role for PARP-1 in the ATN of renal transplantation. We propose the immunohistochemical assessment of PARP-1 in kidney allograft biopsies for early detection of a possible delayed renal function. Copyright: © 2009 O'Valle et al.

Background and objectives: Sinus augmentation is a procedure used for augmenting insufficient bone height that is often observed in the maxillary posterior areas. Many different techniques as well as bone graft regimens have been suggested for performing this procedure. It was the goal of this study to compare, clinically and histologically, two different composite grafting regimens used for sinus augmentation. Material and methods: Five patients, needing a bilateral sinus augmentation to allow implant placement, were recruited for this study. Right sinuses were grafted with cortical bone (collected from overlying the sinus membrane) and bovine hydroxyapatite (HA), while the left side sinuses were grafted with overlying autologous bone plus a bioglass (BG) material. Bone core biopsies were taken at 6 months after sinus graft or at the time of implant insertion. A waiting period of 6 additional months was granted to allow healing, before prosthetic restoration and functional loading. The level of peri-implant bone was evaluated 12 months after loading. A comparative histomorphometric analysis was conducted and a statistical analysis was performed. Results: All implants in both groups were functional after a 12-month loading period. No bone loss was observed radiographically or clinically in both groups. Histologic analysis revealed that both composite grafts had a high biocompatibility. In the bovine HAcontaining group, minimal xenogenic graft absorption was noted. In contrast, BG group samples presented a high absorption rate with some remaining particles imbedded in new normal bone. Conclusions: Sinus augmentation using a combination of autogenous bone plus either bovine HA or BG is a predictable technique.

Objective Vasoactive intestinal peptide (VIP) has been shown to exert potent immunomodulatory activity, and the use of lentiviral vectors has been found to be an effective means of gene delivery. The present study was therefore undertaken to investigate the feasibility and efficiency of gene therapy using lentiviral vectors expressing VIP (LentiVIP) for the treatment of rheumatoid arthritis (RA). Methods We evaluated the therapeutic potential of the gene therapy strategy in the collagen-induced arthritis (CIA) mouse model, administrating the vectors at different phases of the disease. The inflammatory response was determined by measuring the levels of various inflammatory cytokines and chemokines in the joints and serum. The Th1-mediated response was evaluated by determining the proliferative response and cytokine profile of T cells stimulated with autoantigen. Results A single intraperitoneal injection of LentiVIP was highly effective in treating CIA. Mice with established, severe arthritis showed complete regression of the disease. The therapeutic effect of LentiVIP was associated with widespread biodistribution of the vector and increased VIP levels, especially in joints and lymphoid organs, and was mediated through a striking reduction of the 2 deleterious components of the disease, i.e., the autoimmune response (self-reactive Th1 cell activity and autoantibody production) and the inflammatory response. LentiVIP treatment also induced the generation and/or activation of CD4+,CD25+,FoxP3+ Treg cells in arthritic mice. Conclusion Our findings show that in vivo administration of lentiviral vector expressing VIP produces one of the most potent therapeutic effects described so far in any animal model of RA. We propose that VIP gene transfer should be further investigated as a potential novel, effective treatment of RA and other chronic autoimmune disorders.

Red wine polyphenols (RWPs) have been reported to prevent hypertension and endothelial dysfunction. Several individual RWPs exert estrogenic effects. We analyzed the possible in vivo protective effects on blood pressure and endothelial function of RWPs in female spontaneously hypertensive rats (SHR) and its relationship with ovarian function. RWPs (40 mg/kg by gavage) were orally administered for 5 weeks. Ovariectomized rats showed both increased isoprostaglandin F 2α excretion and aortic superoxide production and reduced relaxant response to acetylcholine and contraction to the endothelial nitric oxide synthase (eNOS) inhibitor l -NAME measured in the aorta but similar blood pressure, as compared with sham-operated rats. Moreover, in ovariectomized rats aortic eNOS expression was unchanged, whereas caveolin-1, angiotensin II receptor (AT)-1, and the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunits p22 phox and p47 phox expression was increased compared with sham-operated rats. In both ovariectomized and sham-operated SHR, RWPs reduced systolic blood pressure, urinary isoprostaglandin F 2α excretion, and aortic O 2 − production, improving the endothelium-dependent relaxant response to acetylcholine in SHR. These changes were associated with unchanged aortic eNOS expression, whereas caveolin-1 was increased and the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunits p22 phox and p47 phox expression was reduced. RWPs had no effect on the AT-1 overexpression found in ovariectomized animals. All these results suggest that a chronic treatment with RWPs reduces hypertension and vascular dysfunction through reduction in vascular oxidative stress in female SHR in a manner independent of the ovarian function.

Patients with a protocol renal allograft biopsy simultaneously displaying interstitial fibrosis/tubular atrophy (IF/TA) and subclinical rejection (SCR) have a shortened graft survival than patients with a normal biopsy, or with a biopsy only displaying IF/TA or SCR. The poor outcome of these patients could be related with a more severe inflammation. We evaluate the immunophenotype of infiltrating cells in these diagnostic categories. Nonexhausted paraffin blocks from protocol biopsies done during the first year were stained with anti-CD45, CD3, CD20, CD68 and CD15 monoclonal antibodies. Glomerular and interstitial positive cells were counted. C4d deposition in peritubular capillaries was evaluated. Histological diagnoses were: normal (n = 80), SCR (n = 17), IF/TA (n = 42) and IF/TA + SCR (n = 17). Only interstitial CD20 positive cells were significantly increased in patients displaying IF/TA + SCR; normal (137 +/- 117), SCR (202 +/- 145), IF/TA (208 +/- 151) and IF/TA + SCR (307 +/- 180 cells/mm(2)), p0.01. The proportion of biopsies displaying C4d deposition was not different among groups. The upper tertile of CD20 positive interstitial cells was associated with a decreased death-censored graft survival (relative risk: 3.01, 95% confidence interval: 1.23-7.35; p = 0.015). These data suggest that B-cell interstitial infiltrates are associated with histological damage and outcome, but do not distinguish whether these infiltrates were the cause or the consequence of chronic tubulo-interstitial damage.

The prevalence of subclinical rejection is lower in patients receiving tacrolimus than in patients treated with cyclosporine. However, it is not known whether this difference is related to the modulation of a specific cell immunophenotype. We perform a two case-one control study in patients treated with tacrolimus (n=44) or cyclosporine (n=22) with a protocol biopsy performed at 4 to 6 months. Immunophenotype of infiltrating cells was evaluated with monoclonal antibodies directed against CD45 (all leukocytes), CD3 (T lymphocytes), CD68 (monocytes/macrophages), and CD20 (B lymphocytes) and expressed as interstitial positive cells/mm(2). The number of interstitial CD45 (290+/-209 vs. 696+/-560; P

Objectives: The aim of this cross-sectional study was to evaluate the periodontal status and oral microbiological patterns of a population with end-stage renal disease (ESRD), undergoing haemodialysis (HD). Design: This was a cross-sectional study, involving 52 patients from the Nephrology Department and 52 matched control subjects. Materials and methods: The subjects had a periodontal clinical examination; subgingival plaque samples were taken and analysed using a semiquantitative polymerase chain reaction (PCR) test to detect Porphyromas gingivalis, Tannerella forsythia, Prevotella intermedia, Prevotella nigrescens and Actinobacillus actinomycetemcomitans. Subgingival plaque and saliva samples were studied for Candida and Enterobacteriaceae. Main outcome measures: Most of the 104 subjects had some degree of loss of periodontal attachment (LPA) ≥3 mm [11 (10.5%) had severe LPA; 16 (15.4%) moderate LPA; and 64 (61.5%) mild LPA]. Only 13 subjects (12.5%) presented good periodontal health. Results: No statistically significant differences were found between the HD patients and the control group regarding bleeding index, number of teeth, or percentage of LPA ≥3 mm. However, a statistically significant difference was seen in the degree of oral hygiene. Conclusions: On the basis of the findings presented here, we cannot associate ESRD with more severe periodontal destruction. Although HD patients presented a higher number of periodontopathic microorganisms than the matched controls, a prolonged duration of HD did not bear a statistically significant relationship with the percentage of sites with LPA ≥3 mm, specific microbiota or composition of biofilm.

Objective. To investigate the potential therapeutic action of the immunomodulatory neuropeptide urocortin (UCN) in an experimental model of rheumatoid arthritis (RA). Methods. After disease onset, DBA/1J mice with collagen-induced arthritis (CIA) were treated with UCN, and the incidence, severity (clinical score), and joint histopathology were evaluated. The inflammatory response was determined by measuring the levels of different mediators of inflammation (cytokines and chemokines) in the joints and sera. The Th1-mediated autoreactive response was evaluated by determining the proliferative response and cytokine profile of draining lymph node cells stimulated with the autoantigen and by assaying the content of serum autoantibodies. The number of regulatory CD4+,CD25+ T cells and their capacity to suppress self-reactive Th1 cells were determined in joints and lymph nodes. Results. UCN treatment significantly reduced the incidence and severity of CIA, completely abrogating joint swelling and cartilage and bone destruction. The therapeutic effect of UCN was associated with a striking reduction of the 2 deleterious components of the disease: the Th1-driven autoimmune response and the inflammatory response. UCN also induced the generation and/or activation of efficient interleukin-10/transforming growth factor β1-producing Treg cells in arthritis with the capacity to suppress the autoreactive response and to restore immune tolerance, thus playing a pivotal role in the therapeutic effect of UCN. Conclusion. Our findings provide a powerful rationale for assessing the efficacy of UCN as a novel multistep therapeutic approach to the treatment of RA in humans. © 2007, American College of Rheumatology.

Red wine polyphenols (RWPs) have been reported to exert beneficial effects in preventing cardiovascular diseases, such as hypertension. We studied the effects of chronic treatment with RWPs and apocynin, an inhibitor of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, on blood pressure, endothelial function, and oxidative status in deoxycorticosterone acetate (DOCA)-salt-induced hypertension. Rats were administered RWPs (40 mg/kg) or apocynin (33 microg/kg) daily by gavage for 5 weeks. Plasma catechin levels were detected only after RWP treatment. RWPs and apocynin prevented both the increase in systolic blood pressure and the proteinuria induced by DOCA-salt. Plasma malonyldialdehyde levels, urinary iso-prostaglandin F(2alpha) excretion, aortic superoxide production, and aortic NADPH oxidase activity were found to be increased in animals of the DOCA group. RWP and apocynin treatments reduced these parameters in DOCA-salt rats, having no effect on control rats. However, only RWPs reduced the increase in plasma endothelin-1 (ET-1) levels and aortic p22(phox) gene overexpression found in DOCA-salt animals. RWPs and apocynin also improved the blunted endothelium-dependent relaxation response to acetylcholine in noradrenaline-precontracted aortic rings. All these results suggest that chronic treatment with RWPs prevents hypertension and vascular dysfunction. RWPs prevent vascular oxidative stress by inhibiting NADPH oxidase activity and/or by reducing ET-1 release.

Kidney allografts undergo pretransplant cold ischemia and consequent ischemia-reperfusion injury (IR). Poly (ADP-Ribose) polymerase (PARP-1) overactivation leads to massive NAD consumption and ATP depletion with induction of cellular necrosis under ischemic conditions, which may lead to an increase in acute tubular necrosis (ATN) and a delay in total recovery of renal function (RFR) of the transplanted organ. Materials and Methods. Nuclear PARP-1 immunohistochemical expression (clone: PARP01) was studied in 155 paraffin-embedded renal biopsies from suboptimal donors and 95 kidney allograft biopsies with histopathological diagnosis of ATN. Results. In 50% of ATN biopsies, more than 50% of tubular nuclei were immunostained for PARP-1. PARP-1 expression was higher in ATN biopsies than in those from suboptimal donors (2.40 0.74 vs 0.92 1.13, P 0.0001 Mann-Whitney). PARP-1 showed a statistically significant relationship with the time required to achieve effective diuresis (Rho:0.779), with serum creatinine, and with duration of cold ischemia (Rho: 0.803). These relationships were stronger in the biopsies with ATN. In conclusion, multivariate analysis demonstrated that PARP-1 expression and cold ischemia duration in kidney biopsies with ATN predicted the short-term delay in total recovery of renal function and serum creatinine in the first month.