RESUMO
BACKGROUND: Although kidney transplantation (KT) has become the standard of care for people living with HIV (PLWH) suffering from renal failure, early experiences revealed unanticipated higher rejection rates than those observed in HIV- recipients. The cause of increased acute rejection (AR) in PLWH was assessed by performing a transcriptomic analysis of biopsy specimens, comparing HIV+ to HIV- recipients. METHODS: An analysis of 68 (34 HIV+, 34 HIV-) formalin-fixed paraffin-embedded (FFPE) renal biopsies matched for degree of inflammation was performed from KT recipients with acute T cell-mediated rejection (aTCMR), borderline for aTCMR (BL), and normal findings. Gene expression was measured using the NanoString platform on a custom gene panel to assess differential gene expression (DE) and pathway analysis (PA). RESULTS: DE analysis revealed multiple genes with significantly increased expression in the HIV+ cohort in aTCMR and BL relative to the HIV- cohort. PA of these genes showed enrichment of various inflammatory pathways, particularly innate immune pathways associated with Toll-like receptors. CONCLUSIONS: Upregulation of the innate immune pathways in the biopsies of PLWH with aTCMR and BL is suggestive of a unique immune response that may stem from immune dysregulation related to HIV infection. These findings suggest that these unique HIV-driven pathways may in part be contributory to the increased incidence of allograft rejection after renal transplantation in PLWH.
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Xenobiotics never appear as single, isolated substances in the environment but instead as multi-component mixtures. However, our understanding of the ecotoxicology of mixtures is far from sufficient. In this study, three active pharmaceutical ingredients (carbamazepine, diclofenac, and ibuprofen) and three pesticides (S-metolachlor, terbuthylazine, and tebuconazole) from the most frequently detected emerging micropollutants were examined for their acute cytotoxicity, both individually and in combination, by bioluminescence inhibition in Aliivibrio fischeri (NRRL B-11177). Synergy, additive effects, and antagonism on cytotoxicity were determined using the combination index (CI) method. Additionally, PERMANOVA was performed to reveal the roles of these chemicals in binary, ternary, quaternary, quinary, and senary mixtures influencing the joint effects. Statistical analysis revealed a synergistic effect of diclofenac and carbamazepine, both individually and in combination within the mixtures. Diclofenac also exhibited synergy with S-metolachlor and when mixed with ibuprofen and S-metolachlor. S-metolachlor, whether alone or paired with ibuprofen or diclofenac, increased the toxicity at lower effective concentrations in the mixtures. Non-toxic terbuthylazine showed great toxicity-enhancing ability, especially at low concentrations. Several combinations displayed synergistic effects at environmentally relevant concentrations. The application of PERMANOVA was proven to be unique and successful in determining the roles of compounds in synergistic, additive, and antagonistic effects in mixtures at different effective concentrations.
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Long-term kidney transplant (KT) allograft outcomes have not improved as expected despite a better understanding of rejection and improved immunosuppression. Previous work had validated a computed rejection score, the tissue common rejection module (tCRM), measured by amplification-based assessment of 11 genes from formalin-fixed paraffin-embedded (FFPE) biopsy specimens, which allows for quantitative, unbiased assessment of immune injury. We applied tCRM in a prospective trial of 124 KT recipients, and contrasted assessment by tCRM and histology reads from 2 independent pathologists on protocol and cause biopsies post-transplant. Four 10-µm shaves from FFPE biopsy specimens were used for RNA extraction and amplification by qPCR of the 11 tCRM genes, from which the tCRM score was calculated. Biopsy diagnoses of either acute rejection (AR) or borderline rejection (BL) were considered to have inflammation present, while stable biopsies had no inflammation. Of the 77 biopsies that were read by both pathologists, a total of 40 mismatches in the diagnosis were present. The median tCRM scores for AR, BL, and stable diagnoses were 4.87, 1.85, and 1.27, respectively, with an overall significant difference among all histologic groups (Kruskal-Wallis, p < 0.0001). There were significant differences in tCRM scores between pathologists both finding inflammation vs. disagreement (p = 0.003), and both finding inflammation vs. both finding no inflammation (p < 0.001), along with overall significance between all scores (Kruskal-Wallis, p < 0.001). A logistic regression model predicting graft inflammation using various clinical predictor variables and tCRM revealed the tCRM score as the only significant predictor of graft inflammation (OR: 1.90, 95% CI: 1.40-2.68, p < 0.0001). Accurate, quantitative, and unbiased assessment of rejection of the clinical sample is critical. Given the discrepant diagnoses between pathologists on the same samples, individuals could utilize the tCRM score as a tiebreaker in unclear situations. We propose that the tCRM quantitative score can provide unbiased quantification of graft inflammation, and its rapid evaluation by PCR on the FFPE shave can become a critical adjunct to help drive clinical decision making and immunosuppression delivery.
Assuntos
Aloenxertos/imunologia , Rejeição de Enxerto/diagnóstico , Rejeição de Enxerto/metabolismo , Terapia de Imunossupressão/métodos , Transplante de Rim , Biomarcadores/metabolismo , Biópsia , Feminino , Rejeição de Enxerto/genética , Rejeição de Enxerto/imunologia , Sobrevivência de Enxerto/imunologia , Humanos , Inflamação/genética , Inflamação/metabolismo , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Transcriptoma/genética , Transplante HomólogoRESUMO
BACKGROUND: The role of the soluble urokinase plasminogen activator receptor (suPAR) in focal segmental glomerulosclerosis (FSGS) as the circulating factor or as a predictor of recurrence after transplantation remains controversial. Previously published studies in mice and isolated podocytes produced conflicting results on the effect of suPAR on podocyte injury, effacement of foot processes, and proteinuria. These discordant results were in part due to diverse experimental designs and different strains of mice. The aim of our study was to determine the reasons for the inconsistencies of the previous studies results with suPAR by using uniform methods and studies in different strains of mice. METHODS: We utilized a primary culture of human podocytes and 2 mouse models, the wild type (WT) and the urokinase plasminogen activator receptor (uPAR) KO (uPAR), in an attempt to resolve the reported conflicting results. RESULTS: In both WT and uPAR mouse models, injection of recombinant uPAR, even at a high dose (100 µg), did not induce proteinuria, effacement of podocytes, or disruption of the cytoskeleton. Injection of suPAR resulted in its deposition exclusively in the glomerular endothelial cells and not in the podocytes of WT mice and was not detected at the uPAR KO mice. Kidneys from patients with recurrent FSGS had negative immunostaining for uPAR. We also evaluated the effect of recombinant uPAR on primary culture of human podocytes. uPAR did not result in podocytes damage. CONCLUSIONS: suPAR by itself is not the cause for direct podocyte injury, in vitro or in vivo. These findings suggest a more complex and still poorly understood role of suPAR in FSGS.
Assuntos
Glomerulosclerose Segmentar e Focal/imunologia , Podócitos/patologia , Receptores de Ativador de Plasminogênio Tipo Uroquinase/imunologia , Animais , Autoanticorpos/administração & dosagem , Autoanticorpos/imunologia , Biópsia , Antígenos CD40/imunologia , Células Cultivadas , Glomerulosclerose Segmentar e Focal/sangue , Glomerulosclerose Segmentar e Focal/patologia , Humanos , Rim/citologia , Rim/patologia , Camundongos , Camundongos Knockout , Podócitos/imunologia , Cultura Primária de Células , Receptores de Ativador de Plasminogênio Tipo Uroquinase/administração & dosagem , Receptores de Ativador de Plasminogênio Tipo Uroquinase/genética , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/imunologiaRESUMO
The aim of the present study was to investigate the participation of corticotropin-releasing factor (CRF) receptors (CRF1 and CRF2) in the alterations of the dorsal and ventral striatal dopamine release and the vertical and horizontal locomotor activity observed in rats following chronic nicotine treatment and consequent acute withdrawal. In this purpose, male Wistar rats were exposed to repeated intraperitoneal (ip) injection with nicotine or saline solution for 7â¯days. On the 8th day or the 9th day the rats were injected intracerebroventricularly (icv) with selective CRF1 antagonist antalarmin or selective CRF2 antagonist astressin2B or saline solution. Thirty minutes after the icv injection the changes of the horizontal and vertical locomotor activity were recorded in an in vivo conducta system. Immediately after the behavioral recordings the changes of the dorsal and ventral striatal dopamine release were determined in an in vitro superfusion system. On the 8th day, the horizontal and vertical locomotor activities and the dorsal and ventral striatal dopamine releases increased significantly in nicotine-treated rats, compared to the saline-treated ones. On the 9th day, the horizontal locomotor activity and the dorsal striatal dopamine release increased significantly, whereas the vertical locomotor activity and the ventral striatal dopamine release decreased significantly in nicotine-treated rats, compared to the saline-treated ones. All the changes observed were attenuated significantly by antalarmin, but not astressin2B. The present study demonstrates that the changes of striatal dopamine release and locomotor activity observed following chronic nicotine treatment and consequent acute withdrawal are mediated by CRF1, but not CRF2, receptor.
Assuntos
Neurônios Dopaminérgicos/metabolismo , Nicotina/metabolismo , Receptores de Hormônio Liberador da Corticotropina/metabolismo , Animais , Corpo Estriado/metabolismo , Hormônio Liberador da Corticotropina/farmacologia , Dopamina/metabolismo , Locomoção/fisiologia , Masculino , Atividade Motora , Fragmentos de Peptídeos/farmacologia , Ratos , Ratos Wistar , Síndrome de Abstinência a Substâncias/metabolismoRESUMO
Kisspeptin, a hypothalamic neuropeptide, is a member of the RF-amide family, which have been known to modify pain sensitivity in rodents. The aim of the present study was to investigate the effect of kisspeptin-13 (KP-13), an endogenous derivative of kisspeptin, on nociception in adult male and female CFLP mice and the possible interaction of KP-13 with morphine on nociception. Mice were injected with different doses of KP-13, 30, 60 and 120â¯min after of which the nociceptive sensitivity were assessed via the tail-flick test. To investigate the receptor involved in the mediation a kisspeptin receptor antagonist (KP-234) pretreatment was applied before KP-13 administration. Furthermore, we investigated the effect of KP-13 on the acute antinociceptive effect of morphine, on acute morphine tolerance and on naloxone-precipitated withdrawal. Last, the Von Frey test was used in order to assess KP-13's effect on mechanical nociception. Our results showed that KP-13 decreased the nociceptive threshold of both males and females independent of sex, which was prevented by KP-234. Furthermore, KP-13 treatment depressed the acute antinociceptive effect of morphine and attenuated the development of morphine tolerance. KP-13 also induced a mechanical hypersensitivity. These data underlie kisspeptin's hyperalgesic action and argues for the role of kisspeptin receptor 1 in the mediation of its action. Furthermore, our results suggest that central KP-13 administration can modify the acute effects of morphine.
Assuntos
Tolerância a Medicamentos/genética , Kisspeptinas/genética , Nociceptividade/efeitos dos fármacos , Dor/tratamento farmacológico , Analgésicos Opioides/administração & dosagem , Animais , Kisspeptinas/antagonistas & inibidores , Camundongos , Morfina/administração & dosagem , Nociceptividade/fisiologia , Dor/genética , Dor/fisiopatologia , Limiar da Dor/efeitos dos fármacos , Receptores de Kisspeptina-1/antagonistas & inibidoresRESUMO
The aim of the present study was to investigate the potential anxiolytic- and antidepressant-like actions of Urocortin 2 (Ucn2) and its two fragments, Ucn2 (1-21) and Ucn2 (22-38), in mice, in an attempt to identify the biologically active sequence of this 38 amino acid neuropeptide. In this purpose, male C57BL/6 mice were treated intracerebroventricularly (icv) with 0.125, 0.25, 0.5 and 1⯵g/2⯵l of Ucn2, Ucn2 (1-21) or Ucn2 (22-38). After 30â¯min, the mice were evaluated in an elevated plus-maze test and a forced swim test for anxiety- and depression-like behavior, respectively. Each test lasted 5â¯min. Ucn2 at dose of 0.25⯵g/2⯵l and Ucn2 (1-21) at dose of 0.125⯵g/2⯵l, but not Ucn2 (22-38), increased significantly the number of entries into and the time spent in the open-arms, without influencing the total number of entries. In parallel, the same doses of Ucn2 and Ucn2 (1-21), but not Ucn2 (22-38), increased significantly the climbing and the swimming activity, while decreasing significantly the time of immobility. In addition, Ucn2 at doses of 0.125⯵g/2⯵l and 0.5⯵g/2⯵l decreased significantly the time of immobility, but they did not change the other parameters. The present study demonstrates that Ucn2 exerts anxiolytic- and antidepressant-like effects in C57BL/6 mice, which are mediated by the N-terminal, but not the C-terminal fragment of the peptide. The establishment of the smallest active sequence by further fragmentation of Ucn2 (1-21) may allow the synthesis of new anxiolytic and antidepressant drugs.
Assuntos
Ansiolíticos/uso terapêutico , Antidepressivos/uso terapêutico , Ansiedade/tratamento farmacológico , Depressão/tratamento farmacológico , Urocortinas/uso terapêutico , Animais , Ansiedade/fisiopatologia , Depressão/fisiopatologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Reação de Congelamento Cataléptica/efeitos dos fármacos , Injeções Intraventriculares , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Peptídeos/uso terapêutico , Natação/psicologia , Urocortinas/químicaRESUMO
Hyperglycemia, hyperlipidemia, and free radicals result in platelet activation and atherogenesis. Kisspeptin (KP) is able to regulate metabolism, hemostasis, and the development of atherosclerosis. We examined whether platelet aggregation of streptozotocin-induced diabetic rats depends on the inducer type and if KP-13 and RF-9 (a kisspeptin receptor modifier) can influence platelet function. We measured the speed and the maximum of aggregation, along with the area under the curve. Serum glucose and calcium levels and urine formation of diabetic animals increased, while the body mass and platelet count decreased. Collagen was the most effective inducer of platelet aggregation. The aggregability of nondiabetic platelets was elevated in the presence of 5 × 10-8 mol/L KP-13. This effect was less expressed in diabetic animals. The effectivity of RF-9 was stronger than that of KP-13 in nondiabetic platelets, however it was ineffective in diabetic animals. RF-9 pre-treatment did not change the effects of 5 × 10-8 mol/L KP-13 in either animal group. The in vivo activation of diabetic platelets, which may be due to elevated serum calcium, induces thrombocytopenia and may lead to reduced in vitro aggregability. We could not demonstrate the antagonistic effect of RF-9 against KP-13 in isolated platelets.
Assuntos
Plaquetas/efeitos dos fármacos , Plaquetas/fisiologia , Diabetes Mellitus Experimental/fisiopatologia , Kisspeptinas/farmacologia , Animais , Diabetes Mellitus Experimental/metabolismo , Kisspeptinas/metabolismo , Masculino , Agregação Plaquetária/efeitos dos fármacos , Ratos , Receptores de Kisspeptina-1/metabolismo , Receptores de Neuropeptídeos/metabolismoRESUMO
The aim of the present study was to investigate the effects of the selective agonists of the corticotropin-releasing factor (CRF) 2 receptor, urocortin 2 (UCN 2) and urocortin 3 (UCN 3), on the anxiety- and depression-like signs induced by acute nicotine withdrawal in mice. In order to do so, male CFLP mice were exposed for 7 days to repeated intraperitoneal (IP) injection with nicotine or saline solution and 1day of acute withdrawal and then a single intracerebroventricular (ICV) injection with UCN 2, UCN 3 or saline solution. After 30min the mice were observed in an elevated plus-maze test or a forced swim test, for anxiety- and depression-like behavior. After 5min of testing, the plasma corticosterone concentration reflecting the activity of the hypothalamic-pituitary-adrenal (HPA) axis was also determined by a chemo-fluorescent method. Half of the animals were treated ICV and evaluated on the 8th day, the other half on the 9th day. On the 8th day, nicotine-treated mice presented signs of anxiolysis and depression, but no significant elevation of the plasma corticosterone concentration. On the 9th day, nicotine-treated mice exhibited signs of anxiety and depression and a significant increase of the plasma corticosterone levels. Central administration of UCN 2 or UCN 3 ameliorated the anxiety- and depression-like state including the hyperactivity of the HPA axis, developed during acute withdrawal following chronic nicotine treatment. The present study suggests that selective CRF2 receptor agonists could be used as a therapy in nicotine addiction.
Assuntos
Ansiedade/tratamento farmacológico , Depressão/tratamento farmacológico , Psicotrópicos/administração & dosagem , Receptores de Hormônio Liberador da Corticotropina/agonistas , Síndrome de Abstinência a Substâncias/tratamento farmacológico , Tabagismo/tratamento farmacológico , Animais , Ansiedade/etiologia , Ansiedade/metabolismo , Corticosterona/sangue , Depressão/metabolismo , Depressão/patologia , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Sistema Hipotálamo-Hipofisário/metabolismo , Infusões Intraventriculares , Masculino , Camundongos , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , Sistema Hipófise-Suprarrenal/metabolismo , Receptores de Hormônio Liberador da Corticotropina/metabolismo , Síndrome de Abstinência a Substâncias/metabolismo , Síndrome de Abstinência a Substâncias/psicologia , Tabagismo/metabolismo , Tabagismo/psicologia , Urocortinas/administração & dosagemRESUMO
Kisspeptin has been implicated in cardiovascular control. Eicosanoids play a crucial role in the activation of platelets and the regulation of vascular tone. In the present study, we investigated the effect of kisspeptins on eicosanoid synthesis in platelets and aorta in vitro. Platelets and aorta were isolated from Wistar-Kyoto rats. After preincubation with different doses of kisspeptin, samples were incubated with [1-(14)C]arachidonic acid (0.172 pmol/mL) in tissue culture Medium 199. The amount of labeled eicosanoids was measured with liquid scintillation, after separation with overpressure thin-layer chromatography. Kisspeptin-13 stimulated the thromboxane synthesis. The dose-response curve was bell-shaped and the most effective concentration was 2.5 × 10(-8) mol/L, inducing a 27% increase. Lipoxygenase products of platelets displayed a dose-dependent elevation up to the dose of 5 × 10(-8) mol/L. In the aorta, kisspeptin-13 induced a marked elevation in the production of 6-keto-prostaglandin F1α, the stable metabolite of prostacyclin, and lipoxygenase products. Different effects of kisspeptin on cyclooxygenase and lipoxygenase products indicate that beyond intracellular Ca(2+) mobilization, other signaling pathways might also contribute to its actions. Our data suggest that kisspeptin, through the alteration of eicosanoid synthesis in platelets and aorta, may play a physiologic and (or) pathologic role in the regulation of vascular tone.
Assuntos
Eicosanoides/biossíntese , Kisspeptinas/fisiologia , Vasoconstrição/fisiologia , 6-Cetoprostaglandina F1 alfa/biossíntese , Animais , Aorta/efeitos dos fármacos , Aorta/metabolismo , Ácido Araquidônico/metabolismo , Plaquetas/efeitos dos fármacos , Plaquetas/metabolismo , Relação Dose-Resposta a Droga , Kisspeptinas/farmacologia , Masculino , Ratos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Tromboxanos/biossínteseRESUMO
Periodontal ligament stem cells (PDLSCs) provide an important source for tissue regeneration and may become especially useful in the formation of osteogenic seeds. PDLSCs can be cultured, expanded, and differentiated in vitro; thus, they may be applied in the long-term treatment of the defects in the dental regions. Here we studied numerous potential markers allowing the selection of human PDLSCs with a maximum differentiation potential. We followed the expression of the ATP-binding cassette subfamily G member 2 (ABCG2) membrane transporter protein and isolated ABCG2-expressing cells by using a monoclonal antibody, recognizing the transporter at the cell surface in intact cells. The expression of the ABCG2 protein, corresponding to the so-called side-population phenotype in various tissue-derived stem cells, was found to be a useful marker for the selection of PDLSCs with enhanced osteogenic, chondrogenic, and adipogenic differentiation. These findings may have important applications in achieving efficient dental tissue regeneration by using stem cells from extracted teeth.
Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Antígenos de Diferenciação/metabolismo , Diferenciação Celular/fisiologia , Proteínas de Neoplasias/metabolismo , Ligamento Periodontal/metabolismo , Células-Tronco/metabolismo , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Adolescente , Adulto , Células Cultivadas , Feminino , Humanos , Masculino , Ligamento Periodontal/citologia , Células-Tronco/citologiaRESUMO
The orexigenic peptide ghrelin plays a prominent role in the regulation of energy balance and in the mediation of reward mechanisms and reinforcement for addictive drugs, such as nicotine. Nicotine is the principal psychoactive component in tobacco, which is responsible for addiction and relapse of smokers. Nicotine activates the mesencephalic dopaminergic neurons via nicotinic acetylcholine receptors (nAchR). Ghrelin stimulates the dopaminergic neurons via growth hormone secretagogue receptors (GHS-R1A) in the ventral tegmental area and the substantia nigra pars compacta resulting in the release of dopamine in the ventral and dorsal striatum, respectively. In the present study an in vitro superfusion of rat striatal slices was performed, in order to investigate the direct action of ghrelin on the striatal dopamine release and the interaction of ghrelin with nicotine through this neurotransmitter release. Ghrelin increased significantly the dopamine release from the rat striatum following electrical stimulation. This stimulatory effect was reversed by both the selective nAchR antagonist mecamylamine and the selective GHS-R1A antagonist GHRP-6. Nicotine also increased significantly the dopamine release under the same conditions. This stimulatory effect was antagonized by mecamylamine, but not by GHRP-6. Ghrelin further stimulated the nicotine-induced dopamine release and this effect was abolished by mecamylamine and was partially inhibited by GHRP-6. The present results demonstrate that ghrelin stimulates directly the dopamine release and amplifies the nicotine-induced dopamine release in the rat striatum. We presume that striatal cholinergic interneurons also express GHS-R1A, through which ghrelin can amplify the nicotine-induced dopamine release in the striatum. This study provides further evidence of the impact of ghrelin on the mesolimbic and nigrostriatal dopaminergic pathways. It also suggests that ghrelin signaling may serve as a novel pharmacological target for treatment of addictive and neurodegenerative disorders.
Assuntos
Corpo Estriado/efeitos dos fármacos , Dopamina/metabolismo , Grelina/farmacologia , Nicotina/farmacologia , Animais , Corpo Estriado/metabolismo , Masculino , Ratos , Ratos WistarRESUMO
Obestatin is a 23-amino acid gut-derived neuropeptide, encoded by the same gene with ghrelin. The goal of this study was to examine the effects of obestatin on the acute and chronic analgesic actions of morphine and on mild morphine withdrawal. Open-field (OF) and elevated plus maze (EPM) tests were used to assess mild morphine withdrawal-induced behavior changes and the heat-radiant tail-flick assay was used to investigate analgesic actions of morphine. CFLP male mice were treated twice a day with graded doses of morphine in EPM and OF experiments and once a day in tail-flick studies. Obestatin (1.5µg/2µl) was administrated once a day in all experiments. Furthermore, 0.2mg/kg naloxone or saline was administered after the final injection of morphine at a dose of 20mg/kg in EPM and OF. These behavioral parameters were monitored in the OF: the percentage of center ambulation time and distance; whereas in the EPM: the time spent in open arms and the entries into open arms compared to the total time (%OAT) and entries (%OAE). In the OF, obestatin significantly decreased the percentage of time spent in the center in mice undergoing naloxone-precipitated mild morphine withdrawal. EPM results were similar to open field, but obestatin had no significant effect on parameters mentioned above. Besides, obestatin maintained the analgesic effect of morphine 90 and 120min after morphine injection in mice treated with morphine receiving obestatin compared to mice treated with morphine. In tolerance studies, obestatin diminished the analgesic tolerance to morphine on the 5th day. In this study we confirmed that obestatin reversed the effect of mild morphine withdrawal and enhances the analgesic effect of morphine. These data suggest that obestatin may have a role in opioid-induced analgesia and in behavioral responses induced by opioid withdrawal.
Assuntos
Analgésicos Opioides/farmacologia , Morfina/farmacologia , Antagonistas de Entorpecentes/farmacologia , Hormônios Peptídicos/farmacologia , Síndrome de Abstinência a Substâncias/tratamento farmacológico , Animais , Ansiedade/tratamento farmacológico , Avaliação Pré-Clínica de Medicamentos , Tolerância a Medicamentos , Masculino , Camundongos , Naloxona/farmacologia , Nociceptividade/efeitos dos fármacosRESUMO
The orexigenic peptide ghrelin plays a prominent role in the regulation of energy balance and in the mediation of reward processes and reinforcement for addictive drugs, such as nicotine. Nicotine is the principal psychoactive component in tobacco, which is responsible for addiction and relapse of smokers. Ghrelin and nicotine activates the mesolimbicocortical dopaminergic pathways via growth hormone secretagogue receptors (GHS-R1A) and nicotinic acetylcholine receptors (nAchR), respectively, resulting in the release of dopamine in the nucleus accumbens, the amygdala and the prefrontal cortex. In the present study an in vitro superfusion of rat amygdalar slices was performed in order to investigate the direct action of ghrelin and nicotine on the amygdalar dopamine release. Ghrelin increased significantly the dopamine release from the rat amygdala following electrical stimulation. This effect was inhibited by both the selective GHS-R1A antagonist GHRP-6 and the selective nAchR antagonist mecamylamine. Under the same conditions, nicotine also increased significantly the dopamine release from the rat amygdala. This effect was antagonized by mecamylamine, but not by GHRP-6. Co-administration of ghrelin and nicotine induced a similar increase of amygdalar dopamine release. This stimulatory effect was partially reversed by both GHRP-6 and mecamylamine. The present results demonstrate that both ghrelin and nicotine stimulates directly the dopamine release in the amygdala, an important dopaminergic target area of the mesolimbicocortical pathway.
Assuntos
Tonsila do Cerebelo/metabolismo , Dopamina/metabolismo , Grelina/farmacologia , Nicotina/farmacologia , Tonsila do Cerebelo/efeitos dos fármacos , Animais , Técnicas In Vitro , Masculino , Mecamilamina , Oligopeptídeos/farmacologia , Córtex Pré-Frontal/metabolismo , Ratos , Ratos Wistar , Receptores de Grelina/efeitos dos fármacos , Receptores Nicotínicos/efeitos dos fármacosRESUMO
The aim of the present investigation was to study the effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on morphine withdrawal-induced behavioral changes and hypothermia in male CFLP mice. Elevated plus maze (EPM) and jump tests were used to assess naloxone-precipitated morphine withdrawal-induced behavior responses. Different doses of subcutaneous (s.c.) naloxone, (0.1 and 0.2 mg/kg, respectively) were used to precipitate the emotional and psychical aspects of withdrawal on EPM and 1 mg/kg (s.c.) was used to induce the somatic withdrawal signs such as jumping, and the changes in body temperature. In our EPM studies, naloxone proved to be anxiolytic in mice treated with morphine. Chronic intracerebroventricular (i.c.v.) administration of PACAP alone had no significant effect on withdrawal-induced anxiolysis and total activity at doses of 500 ng and 1 µg. At dose of 500 ng, however, PACAP significantly counteracted the reduced motor activity in the EPM test in mice treated with morphine and diminished the hypothermia and shortened jump latency induced by naloxone in mice treated with morphine. These findings indicate that anxiolytic-like behavior may be mediated via a PACAP-involved pathway and PACAP may play an important role in chronic morphine withdrawal-induced hypothermia as well.
Assuntos
Hipotermia/tratamento farmacológico , Aprendizagem em Labirinto/efeitos dos fármacos , Morfina/toxicidade , Atividade Motora/efeitos dos fármacos , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/uso terapêutico , Síndrome de Abstinência a Substâncias/tratamento farmacológico , Animais , Hipotermia/induzido quimicamente , Masculino , Camundongos , Naloxona/farmacologia , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/farmacologiaRESUMO
OBJECTIVES: To investigate salivary proteins with proteomic technologies to evaluate protein composition differences between samples with cleft lip and palate and healthy controls. DESIGN, PARTICIPANTS: Matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI TOF/TOF) mass spectrometry was used as a high-throughput analytical technique for identification of nonsyndromic cleft lip and palate stimulated salivary proteins. The samples consisted of two groups: 31 cleft lip and palate patients and a control group with 20 healthy volunteers. RESULTS: The presence of cleft lip and palate stimulated the expression of several proteins, included adaptor-related protein complex 3, dermokine, nidogen 1 precursor, transforming growth factor-ß3, and a zinc finger RAN-binding domain containing 2. CONCLUSIONS: The salivary proteome of cleft lip and palate patients differs from the protein composition of healthy control saliva samples. Several common secreted proteins such as actins, salivary cystatins, and keratins were upregulated by cleft; increased levels of TGF-ß3 and dermokine were detected in the pathologic samples. The current proteomic results suggest keratinocyte activation among patients with cleft lip and palate. The score of our preliminary results suggests the hypothesis that identified salivary proteins are of vital clinical importance in tissue regeneration and the molecular repair mechanism seen in patients with cleft lip and palate.
Assuntos
Fenda Labial/metabolismo , Fissura Palatina/metabolismo , Proteômica , Proteínas e Peptídeos Salivares/metabolismo , Estudos de Casos e Controles , Criança , Pré-Escolar , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Masculino , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Regulação para CimaRESUMO
OBJECTIVE: The aim of the study was to estimate the accuracy of panoramic radiographic signs predicting inferior alveolar nerve (IAN) paresthesia after lower third molar removal. STUDY DESIGN: In a case-control study the sample was composed of 41 cases with postoperative IAN paresthesia and 359 control cases without it. The collected data included "classic" specific signs indicating a close spatial relationship between third molar root and inferior alveolar canal (IAC), root curvatures, and the extent of IAC-root tip overlap. Bivariate and multivariate logistic regression analyses were completed to estimate the association between radiographic findings and IAN paresthesia. RESULTS: The multivariate logistic analysis identified 3 signs significantly associated with IAN paresthesia (P < .001): interruption of the superior cortex of the canal wall, diversion of the canal, and darkening of the root. The sensitivities and specificities ranged from 14.6% to 68.3% and from 85.5% to 96.9%, respectively. The positive predictive values, calculated to factor a 1.1% prevalence of paresthesia, ranged from 3.6% to 10.9%, whereas the negative predictive values >99%. CONCLUSION: Panoramic radiography is an inadequate screening method for predicting IAN paresthesia after mandibular third molar removal.
Assuntos
Nervo Mandibular/diagnóstico por imagem , Dente Serotino/cirurgia , Radiografia Panorâmica , Extração Dentária/efeitos adversos , Traumatismos do Nervo Trigêmeo , Adulto , Idoso , Distribuição de Qui-Quadrado , Traumatismos dos Nervos Cranianos/diagnóstico por imagem , Traumatismos dos Nervos Cranianos/etiologia , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Parestesia/etiologia , Prognóstico , Estudos Retrospectivos , Sensibilidade e Especificidade , Estatísticas não Paramétricas , Raiz Dentária/diagnóstico por imagem , Adulto JovemRESUMO
The endomorphins (EM1: Tyr-Pro-Trp-Phe-NH2, and EM2: Tyr-Pro-Phe-Phe-NH2) are recently discovered endogenous ligands for mu-opioid receptors (MORs) with role of neurotransmitters or neuromodulators in mammals. Cessation of their physiological action may be effected through rapid enzymatic degradation by the dipeptidyl-peptidase IV (DPPIV) found in the brain synaptic membranes. An in vitro superfusion system was utilized to investigate the actions of EM1, EM2 and specific DPPIV inhibitor diprotin A on the striatal release of dopamine (DA) induced by electrical stimulation in rats. The involvement of the different MORs (MOR1 and MOR2) in this process was studied by pretreatment with MOR antagonists beta-funaltrexamine (a MOR1 and MOR2 antagonist) and naloxonazine (a MOR1 antagonist). EM1 significantly increased the tritium-labelled dopamine DA release induced by electrical stimulation. EM2 was effective only when the slices were pretreated with diprotin A. beta-Funaltrexamine antagonized the stimulatory effects of both EM1 and EM2. The administration of naloxonazine did not appreciably influence the action of EM1, but blocked the action of EM2, at least when the slices were pretreated with diprotin A. These data suggest that both EM1 and EM2 increase DA release from the striatum and, though diprotin A does not affect the action of EM1, it inhibits the enzymatic degradation of EM2. The DA-stimulating action induced by EM1 seems to be mediated by MOR2, while that evoked by EM2 appears to be transmitted by MOR1.
Assuntos
Corpo Estriado/efeitos dos fármacos , Inibidores da Dipeptidil Peptidase IV , Dopamina/metabolismo , Oligopeptídeos/antagonistas & inibidores , Oligopeptídeos/farmacologia , Terminações Pré-Sinápticas/efeitos dos fármacos , Animais , Corpo Estriado/metabolismo , Dipeptidil Peptidase 4/metabolismo , Estimulação Elétrica , Bombas de Infusão , Masculino , Naloxona/análogos & derivados , Naloxona/farmacologia , Naltrexona/análogos & derivados , Naltrexona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Vias Neurais/efeitos dos fármacos , Vias Neurais/metabolismo , Oligopeptídeos/metabolismo , Técnicas de Cultura de Órgãos , Terminações Pré-Sinápticas/metabolismo , Ensaio Radioligante/métodos , Ratos , Ratos Wistar , Substância Negra/efeitos dos fármacos , Substância Negra/metabolismo , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologiaRESUMO
Oral health-related quality of life (OHRQoL) is increasingly of interest in dentistry. The current internationally accepted instruments used to measure OHRQoL need to be cross-culturally adapted for use in other cultural environments. It was the aim of the present study to develop a Hungarian version of the Oral Health Impact Profile (OHIP-H) following accepted guidelines. The original English-language version was translated into Hungarian, back-translated into English, and tested for its psychometric properties. Construct validity was tested on 144 prosthodontic patients and 200 randomly selected subjects. A priori hypothesized associations between OHIP summary scores and self-reported oral health and six self-reported oral conditions were investigated. The pattern of the observed associations supported the new instrument's construct validity. Responsiveness tested in 28 oral surgery patients was indicated by a statistically significant mean OHIP score change from 39.2 to 23.0. Test-retest reliability was demonstrated by intraclass correlation coefficients of 0.81-0.90 for OHIP summary scores and subscales in 31 prosthodontic patients. Cronbach's alpha values between 0.71 and 0.96 proved to have high internal consistency. Adequate psychometric properties in typical patient populations make the new instrument suitable for assessment of OHRQoL in Hungary.
Assuntos
Atitude Frente a Saúde , Saúde Bucal , Qualidade de Vida , Inquéritos e Questionários , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Bruxismo/psicologia , Síndrome da Ardência Bucal/psicologia , Criança , Comparação Transcultural , Dentaduras/psicologia , Feminino , Halitose/psicologia , Humanos , Hungria , Idioma , Masculino , Pessoa de Meia-Idade , Psicometria/estatística & dados numéricos , Reprodutibilidade dos Testes , Transtornos da Articulação Temporomandibular/psicologiaRESUMO
The rapid progression of digital radiography makes us consider its role in different fields of dentistry. The aim of the present survey was to evaluate the advantages of digital imaging over the use of radiographic films in the diagnosis and therapy of common oral pathologic conditions. The underlying hypothesis was that RadioVisiograph (RVG, Trophy Radiology, Marne-la-Vallée, France)--provided that there is access to this digital network--is easily available. Thus it can provide options for specialists of different dental fields to obtain more detailed information in order to make better decisions. Immediately after the installation of the new system and then, ten months later we analysed the results of digital images and radiographic films in outpatients of different sections at our clinic. The quality of both diagnostic procedures was evaluated using a questionnaire. Digital imaging was used mainly by specialists of endodontics, followed by restorative and prosthetic dentistry in setting up the diagnosis. The application of this new technique in parodontology and paediatric dentistry was negligible. However, it is worth mentioning that the majority of dentists participating in the survey thought that conventional radiographic films gave more detailed imaging than the digital technology. To sum it up, currently, digital imaging systems may represent an efficient device in diagnostic and therapeutic procedures. Nevertheless owing to the technical sensitivity of the above systems, traditional radiographic techniques should also be used as an option due to their advantages.