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1.
Braz J Biol ; 84: e263041, 2022.
Article in English | MEDLINE | ID: mdl-35792738

ABSTRACT

Commercial fig tree cultivation in Brazil involves a single cultivar, 'Roxo-de-Valinhos'. The use of a single cultivar results in serious diseases and related problems. The aim of this study was to characterize fig accessions by analyzing the natural root-knot nematode and leaf rust incidence in relation to the epigenomic profile of the plant, since epigenetic variations affect plant-pathogen interactions. All plants were attacked by nematodes, indicating susceptibility; Meloidogyne incognita was the root-knot nematode species involved. Joint analysis of data showed that methylation and leaf rust incidence were correlated when observed in the same phenological phase, presenting initial evidence of the same factorial pressure loads in genotypes, suggesting similar behavior within these genotypes.


Subject(s)
Ficus , Tylenchoidea , Animals , Incidence , Methylation , Plant Roots , Plants , Trees/genetics
2.
Braz J Biol ; 84: e260650, 2022.
Article in English | MEDLINE | ID: mdl-35674587

ABSTRACT

With increased interest in cultivation, the study of white-fleshed pitahaya (Selenicereus undatus (Haw.) D.R. Hunt, Cactaceae family) seedling production is of fundamental importance in the search for novel techniques to increase cultivation and guarantee homogeneous and productive orchards. The present study investigated the influence of various gibberellic acid (GA3) concentrations and fruit maturation stages on seed germination and vigor of white-fleshed pitahaya seedlings, considering the physiological quality of seedlings produced to support genetic breeding and conservation programs of the species. White-fleshed pitahaya seeds at two maturation stages (physiologically ripe and maintained at 10 °C in Biochemical Oxygen Demand incubators for three months) were treated with varying GA3 concentrations of 0, 50, 100, and 500 mg/L. We observed the influence of fruit storage on seedling germination, emergence, and growth as a function of GA3 concentration. According to the results, seeds extracted from ripe white-fleshed pitahaya fruits grown under the conditions tested here required GA3 application to increase seedling emergence and vigor, with optimal doses in the 150-300-mg/L range. In the case of pitahaya fruits intended for storage for future seed removal and maintained under the same sowing conditions, the application of higher doses of GA3 was necessary when compared to the previous condition, with a minimum dose of 500 mg/L GA3. The present study shows that the maturation stage of white-fleshed pitahaya fruits intended for seed removal influences the quality of seedlings; therefore, the use of seeds extracted from ripe pitahaya fruits without fermentation is more appropriate for the purpose.


Subject(s)
Germination , Seedlings , Fruit , Germination/physiology , Gibberellins , Plant Breeding , Seeds
3.
Mol Cell Neurosci ; 105: 103497, 2020 06.
Article in English | MEDLINE | ID: mdl-32353527

ABSTRACT

Various animal models have been employed to understand the pathogenic mechanism of neuropathic pain. Nitric oxide (NO) is an important molecule in nociceptive transmission and is involved in neuropathic pain. However, its mechanistic actions remain unclear. The aim of this study was to better understand the involvement of neuronal and inducible isoforms of nitric oxide synthase (nNOS and iNOS) in neuropathic pain induced by chronic constriction injury (CCI) of the sciatic nerve in rats. We evaluated pain sensitivity (mechanical withdrawal thresholds using Randall and Selitto, and von Frey tests, and thermal withdrawal thresholds using Hargreaves test) prior to CCI surgery, 14 days post CCI and after intrathecal injections of selective nNOS or iNOS inhibitors. We also evaluated the distribution of NOS isozymes in the spinal cord and dorsal root ganglia (DRG) by immunohistochemistry, synthesis of iNOS and nNOS by Western blot, and NO production using fluorescent probe DAF-2 DA (DA). Our results showed higher number of nNOS and iNOS-positive neurons in the spinal cord and DRG of CCI compared to sham rats, and their reduction in CCI rats after treatment with selective inhibitors compared to non-treated groups. Western blot results also indicated reduced expression of nNOS and iNOS after treatment with selective inhibitors. Furthermore, both inhibitors reduced CCI-evoked mechanical and thermal withdrawal thresholds but only nNOS inhibitor was able to efficiently lower mechanical withdrawal thresholds using von Frey test. In addition, we observed higher NO production in the spinal cord and DRG of injured rats compared to control group. Our study innovatively shows that nNOS may strongly modulate nociceptive transmission in rats with neuropathic pain, while iNOS may partially participate in the development of nociceptive responses. Thus, drugs targeting nNOS for neuropathic pain may represent a potential therapeutic strategy.


Subject(s)
Ganglia, Spinal/metabolism , Neuralgia/metabolism , Nitric Oxide/metabolism , Sciatic Nerve/metabolism , Animals , Hyperalgesia/drug therapy , Male , Nitric Oxide Synthase Type II/metabolism , Rats, Wistar , Spinal Cord/metabolism
4.
Brain Res ; 1727: 146521, 2020 01 15.
Article in English | MEDLINE | ID: mdl-31697924

ABSTRACT

In addition to motor symptoms, Parkinson's disease (PD) presents high prevalence of painful symptoms responsible for worsening quality of life of PD patients. Physical exercise can improve such painful symptoms. This study evaluated the effects of exercise on nociceptive threshold using an unilateral rat model of PD, as well as the role played by cannabinoid and opioid receptors in areas responsible for pain pathways. For PD induction, Wistar rats were injected with 6-OHDA. 15 days after, rats either remained sedentary or were forced to exercise three times a week for 40 min. Motor and nociceptive behaviors were evaluated through cylinder and mechanical hyperalgesia tests, respectively. The animals were euthanized for analysis of cannabinoid receptor type 1 (CB1) and type 2 (CB2), and µ-opioid receptor (MOR) in the anterior cingulate cortex (ACC), periaqueductal gray matter (PAG), and thalamus areas by immunohistochemistry (IHC) and Western blotting. Our data revealed a decrease in the nociceptive threshold in both forepaws after surgery; in contrast, there was improvement in painful symptoms after the exercise protocol. For cannabinoid system there were an increase in CB2 expression in the ACC and PAG, and in CB1 levels in the PAG. And for opioid system there was an increase of MOR expression in the thalamus. Thus, modulation of those receptors by physical exercise can be an important non-pharmacological intervention to reduce painful symptoms in a rat model of PD, contributing to knowledge and promotion of better treatment aimed at improving the quality of life of PD patients.


Subject(s)
Nociception/physiology , Parkinson Disease/metabolism , Parkinson Disease/psychology , Physical Conditioning, Animal , Receptor, Cannabinoid, CB1/metabolism , Receptor, Cannabinoid, CB2/metabolism , Receptors, Opioid, mu/metabolism , Animals , Disease Models, Animal , Gyrus Cinguli/metabolism , Hyperalgesia/complications , Hyperalgesia/prevention & control , Parkinson Disease/prevention & control , Periaqueductal Gray/metabolism , Rats, Wistar
5.
Clin Exp Immunol ; 187(3): 383-398, 2017 03.
Article in English | MEDLINE | ID: mdl-28008595

ABSTRACT

Defective apoptosis might be involved in the pathogenesis of multiple sclerosis (MS). We evaluated apoptosis-related molecules in MS patients before and after autologous haematopoietic stem cell transplantation (AHSCT) using BCNU, Etoposide, AraC and Melphalan (BEAM) or cyclophosphamide (CY)-based conditioning regimens. Patients were followed for clinical and immunological parameters for 2 years after AHSCT. At baseline, MS patients had decreased proapoptotic BAD, BAX and FASL and increased A1 gene expression when compared with healthy counterparts. In the BEAM group, BAK, BIK, BIMEL , FAS, FASL, A1, BCL2, BCLXL , CFLIPL and CIAP2 genes were up-regulated after AHSCT. With the exception of BIK, BIMEL and A1, all genes reached levels similar to controls at day + 720 post-transplantation. Furthermore, in these patients, we observed increased CD8+ Fas+ T cell frequencies after AHSCT when compared to baseline. In the CY group, we observed increased BAX, BCLW, CFLIPL and CIAP1 and decreased BIK and BID gene expressions after transplantation. At day + 720 post-AHSCT, the expression of BAX, FAS, FASL, BCL2, BCLXL and CIAP1 was similar to that of controls. Protein analyses showed increased Bcl-2 expression before transplantation. At 1 year post-AHSCT, expression of Bak, Bim, Bcl-2, Bcl-xL and cFlip-L was decreased when compared to baseline values. In summary, our findings suggest that normalization of apoptosis-related molecules is associated with the early therapeutic effects of AHSCT in MS patients. These mechanisms may be involved in the re-establishment of immune tolerance during the first 2 years post-transplantation.


Subject(s)
Apoptosis/genetics , Autophagy-Related Protein 5/genetics , Multiple Sclerosis/genetics , Adult , CD8-Positive T-Lymphocytes/drug effects , Cyclophosphamide/therapeutic use , Female , Gene Expression/genetics , Hematopoietic Stem Cell Transplantation/methods , Humans , Male , Middle Aged , Multiple Sclerosis/drug therapy , Transplantation Conditioning/methods , Transplantation, Autologous/methods , Young Adult
6.
J Clin Pathol ; 69(10): 926-9, 2016 Oct.
Article in English | MEDLINE | ID: mdl-27402956

ABSTRACT

Despite all the knowledge, the cellular and molecular mechanisms involved in myeloproliferative neoplasm (MPN) pathophysiology remain unclear. Authors have shown galectin-1 (Gal-1) and 3 playing roles in tumour angiogenesis and fibrosis, which were correlated with poor prognosis in patients with MPN. In the present study LGALS1 and LGALS3 were differently expressed between polycythemia vera, essential thrombocythemia (ET) and primary myelofibrosis (PMF) diseases. Increased LGALS3 expression was associated with a negative JAK2 V617F status mutation in leucocytes from PMF but not in patients with ET without this mutation. However, a positive Janus kinase 2 (JAK2) V617F cell line established from patients with ET (SET-2 cells) when treated with JAK inhibitor presented high levels of LGALS3. Additionally, high LGALS1 expression was found in CD34(+) cells but not in leucocytes from patients with PMF, in absence of JAK2 V617F mutation, and also in SET-2 cells treated with JAK inhibitor. Thus, our findings indicate that differential expression of LGALS1 and/or LGALS3 in patients with MPN is linked with JAK2 V617F status mutation in these diseases and state of cell differentiation.


Subject(s)
Galectin 1/genetics , Galectin 3/genetics , Janus Kinase 2/genetics , Polycythemia Vera/genetics , Primary Myelofibrosis/genetics , Thrombocythemia, Essential/genetics , Adult , Amino Acid Substitution , Antigens, CD34/genetics , Blood Proteins , Bone Marrow/pathology , Bone Marrow Neoplasms/diagnosis , Bone Marrow Neoplasms/genetics , Bone Marrow Neoplasms/metabolism , Cell Line , Galectin 1/metabolism , Galectin 3/metabolism , Galectins , Gene Expression Regulation, Neoplastic , Humans , Janus Kinase 2/metabolism , Male , Middle Aged , Mutation , Myeloproliferative Disorders/diagnosis , Myeloproliferative Disorders/genetics , Myeloproliferative Disorders/metabolism , Polycythemia Vera/diagnosis , Polycythemia Vera/metabolism , Primary Myelofibrosis/diagnosis , Primary Myelofibrosis/metabolism , Thrombocythemia, Essential/diagnosis , Thrombocythemia, Essential/metabolism
7.
Drug Chem Toxicol ; 39(1): 28-34, 2016.
Article in English | MEDLINE | ID: mdl-25690107

ABSTRACT

Amphotericin B is the "gold standard" agent in the management of serious systemic fungal infections. However, this drug can cause nephrotoxicity, which contributes up to 25% of all acute kidney injuries in critically ill patients. Cyclic adenosine monophosphate can protect kidney cells from death due to injury or drug exposure in some cases. Hence, the objective of this work was to evaluate if cAMP could prevent cell death that occurs in renal cell lines subjected to AmB treatment and, if so, to assess the involvement of PKA in the transduction of this signal. Two different renal cell lines (LLC-PK1 and MDCK) were used in this study. MTT and flow cytometry assays showed increased cell survival when cells were exposed to cAMP in a PKA-independent manner, which was confirmed by western blot. This finding suggests that cAMP (db-cAMP) may prevent cell death caused by exposure to AmB. This is the first time this effect has been identified when renal cells are exposed to AmB's nephrotoxic potential.


Subject(s)
Amphotericin B/toxicity , Antifungal Agents/toxicity , Cyclic AMP/administration & dosage , Kidney/drug effects , Animals , Blotting, Western , Cell Survival/drug effects , Cyclic AMP/metabolism , Cyclic AMP-Dependent Protein Kinases/metabolism , Dogs , Flow Cytometry , Kidney/pathology , LLC-PK1 Cells , Madin Darby Canine Kidney Cells , Signal Transduction/drug effects , Swine
8.
Radiat Prot Dosimetry ; 162(1-2): 52-7, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25009185

ABSTRACT

In this work, the authors revisit the measurements of Trout and Kelley, and Simpkin and Dixon, by means of Monte Carlo simulations. Starting with a simple cylindrical homogeneous phantom, the authors introduce a more realistic phantom and the effect of the bucky in the simulations. The results indicate that optimised shielding methodologies should not neglect the attenuation and scattering by the patient plus bucky.


Subject(s)
Computer Simulation , Image Processing, Computer-Assisted , Monte Carlo Method , Scattering, Radiation , Humans , Phantoms, Imaging , Tomography, Emission-Computed, Single-Photon
9.
Toxicol Mech Methods ; 24(6): 369-76, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24785081

ABSTRACT

Cyclosporine is an important immunosuppressive agent; however, nephrotoxicity is one of the main adverse effects. The purpose of this study was to evaluate the effect of inhibiting the protein kinase A (PKA) signaling pathway in nephrotoxicity caused by cyclosporine from the assessment of cell viability, pro-inflammatory cytokines, and nitric oxide (NO) production in LLC-PK1 and MDCK cell lines. Cyclosporine proved to be cytotoxic for both cell lines, as assessed by the mitochondrial enzyme activity assay (MTT), caused DNA fragmentation, determined by flow cytometry using the propidium iodide dye, and activated the PKA pathway (western blot assay). In MDCK cells, the inhibition of the PKA signaling pathway (H89 inhibitor) caused a significant reduction in DNA fragmentation. In both cell lines, the production of IL-6 proved to be a dependent PKA pathway, while TNF-α was not influenced by the inhibition of the PKA pathway. The NO production was increased when cells were pre-incubated with H89 followed by cyclosporine, and this production was dependent on the PKA pathway in LLC-PK1 and MDCK cells lines. Therefore, considering the present study's results, it can be concluded that the inhibition of PKA signaling pathway can aid in reducing the degree of nephrotoxicity caused by cyclosporine.


Subject(s)
Cyclic AMP-Dependent Protein Kinases/metabolism , Cyclosporine/toxicity , Kidney/drug effects , Signal Transduction/drug effects , Animals , Blotting, Western , Cell Line , Cell Survival , Cyclic AMP-Dependent Protein Kinases/antagonists & inhibitors , Cyclic AMP-Dependent Protein Kinases/genetics , DNA Fragmentation , Dogs , Gene Expression Regulation, Enzymologic/drug effects , Immunosuppressive Agents/toxicity , Isoquinolines/pharmacology , Kidney/cytology , Nitric Oxide/metabolism , Protein Kinase Inhibitors/pharmacology , Sulfonamides/pharmacology , Swine
10.
Blood Cells Mol Dis ; 53(1-2): 47-55, 2014.
Article in English | MEDLINE | ID: mdl-24629639

ABSTRACT

BACKGROUND: Chronic myeloid leukemia (CML) is a myeloproliferative disease characterized by the presence of Philadelphia chromosome (Ph) leading to expression of a BCR-ABL1 fusion oncogene. The BCR-ABL protein has a constitutive tyrosine kinase activity which is responsible for CML pathogenesis by promoting cell apoptosis resistance; however, the cellular and molecular mechanisms associated with BCR-ABL expression and apoptosis impairment in CML leukemic cells have not been fully elucidated. METHODS: This study evaluated apoptomiRs and their predicted apoptotic genes in BCR-ABL(+) cells from patients in different phases of CML treated with tyrosine kinase inhibitor (TKI) according to their imatinib (IM) response by qPCR. Phosphotyrosine and c-ABL expressions in HL-60.BCR-ABL cells treated with TKI were done by Western blot. RESULTS: We found that dasatinib (DAS) modulated miR-let-7d, miR-let-7e, miR-15a, miR-16, miR-21, miR-130a and miR-142-3p expressions while IM modulated miR-15a and miR-130a levels. miR-16, miR-130a and miR-145 expressions were modulated by nilotinib (NIL). We observed higher miR-15a, miR-130b and miR-145; and lower miR-16, miR-26a and miR-146a expressions in CML-CP in comparison with controls. CML-AP patients showed low miR-let-7d, miR-15a, miR-16, miR-29c, miR-142-3p, miR-145, and miR-146a levels in comparison with CML-CP. We noted that the miR-26a, miR-29c, miR-130b and miR-146a expressions were downregulated in IM resistant patients in comparison with IM responsive patients. CONCLUSIONS: This study showed the modulation of apoptomiRs by BCR-ABL kinase activity and the deregulation of apoptomiRs and their predicted apoptotic target genes in different CML phases and after treatment with TK inhibitors. ApoptomiRs may be involved in the BCR-ABL(+) cell apoptosis regulation.


Subject(s)
Apoptosis/genetics , Benzamides/pharmacology , Drug Resistance, Neoplasm/genetics , Fusion Proteins, bcr-abl/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology , MicroRNAs/genetics , Piperazines/pharmacology , Pyrimidines/pharmacology , Adolescent , Adult , Aged , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Benzamides/therapeutic use , Case-Control Studies , Cell Line, Tumor , Disease Progression , Female , Fusion Proteins, bcr-abl/metabolism , Gene Expression Regulation, Leukemic/drug effects , Humans , Imatinib Mesylate , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy , Male , Middle Aged , Neoplasm Staging , Phosphorylation/drug effects , Piperazines/therapeutic use , Protein Kinase Inhibitors/pharmacology , Protein Kinase Inhibitors/therapeutic use , Proto-Oncogene Proteins c-abl/genetics , Proto-Oncogene Proteins c-abl/metabolism , Pyrimidines/therapeutic use , Treatment Outcome , Young Adult
11.
J Appl Toxicol ; 34(12): 1285-92, 2014 Dec.
Article in English | MEDLINE | ID: mdl-24105867

ABSTRACT

Amphotericin B is one of the most effective antifungal agents; however, its use is often limited owing to adverse effects, especially nephrotoxicity. The purpose of this study was to evaluate the effect of inhibiting the PKA signaling pathway in nephrotoxicity using Amphotericin B from the assessment of cell viability, pro-inflammatory cytokines and nitric oxide (NO) production in LLC-PK1 and MDCK cell lines. Amphotericin B proved to be cytotoxic for both cell lines, as assessed by the mitochondrial enzyme activity (MTT) assay; caused DNA fragmentation, determined by flow cytometry using the propidium iodide (PI) dye; and activated the PKA pathway (western blot assay). In MDCK cells, the inhibition of the PKA signaling pathway (using the H89 inhibitor) caused a significant reduction in DNA fragmentation. In both cells lines the production of interleukin-6 (IL)-6 proved to be a dependent PKA pathway, whereas tumor necrosis factor-alpha (TNF-α) was not influenced by the inhibition of the PKA pathway. The NO production was increased when cells were pre-incubated with H89 followed by Amphotericin B, and this production produced a dependent PKA pathway in LLC-PK1 and MDCK cells lines. Therefore, considering the present study's results as a whole, it can be concluded that the inhibition of the PKA signaling pathway can aid in reducing the degree of nephrotoxicity caused by Amphotericin B.


Subject(s)
Amphotericin B/toxicity , Antifungal Agents/toxicity , Cyclic AMP-Dependent Protein Kinases/metabolism , Cytokines/biosynthesis , Kidney/drug effects , Nitric Oxide/biosynthesis , Animals , Cell Culture Techniques , Cell Survival/drug effects , Cyclic AMP-Dependent Protein Kinases/antagonists & inhibitors , DNA Fragmentation/drug effects , Dogs , Interleukin-6/biosynthesis , Kidney/enzymology , Kidney/immunology , Kidney/pathology , LLC-PK1 Cells , Madin Darby Canine Kidney Cells , Signal Transduction , Swine , Tumor Necrosis Factor-alpha/biosynthesis
12.
Arq. bras. med. vet. zootec ; 65(3): 742-748, June 2013. ilus, tab
Article in Portuguese | LILACS | ID: lil-679108

ABSTRACT

Avaliaram-se a incidência de endometrite citológica dos 29 aos 90 dias pós-parto e seus efeitos sobre o desempenho reprodutivo de vacas de corte Nelore submetidas a uma estação de monta (EM) de 90 dias. Foram utilizadas 49 matrizes Nelores, sem histórico de retenção de placenta, sem a presença de uma infecção uterina clínica, e com escore de condição corporal acima de 2,5. Realizou-se exame ultrassonográfico para avaliar a parede uterina e a atividade ovariana. O diagnóstico de endometrite citológica foi feito pela técnica de lavagem uterina, considerando-se caso de endometrite ≥5% de neutrófilos em cada lâmina. A incidência de endometrite citológica do rebanho foi de 22%, não diferindo entre as categorias analisadas (primíparas versus multíparas) (P>0,05), a taxa de concepção à primeira inseminação também foi semelhante entre primíparas versus multíparas (P>0,05), porém a taxa de gestação ao final da EM foi maior nas vacas multíparas (83,8%) quando comparadas às primíparas (50,0%) (P<0,05). A presença ou ausência da endometrite citológica não influenciou a taxa de concepção (P>0,05), tampouco a taxa de gestação ao final da EM (P>0,05). Conclui-se que o uso da citologia endometrial não se justifica como ferramenta de diagnóstico em vacas de corte Nelore.


Were evaluated the incidence of cytological endometritis from 29 to 90 days postpartum and its effect on the reproductive performance of Nelore beef cows submitted to a breeding season (BS) for 90 days. A total of 49 cows, with no history of retained placenta, without the presence of a clinic uterine infection, and with a body condition score above 2.5 were used. Ultrasound examination was performed to evaluate the uterine wall and ovarian activity. The cytological diagnosis of endometritis was done by uterine lavage, and endometritis was considering cases of ≥5% neutrophils in each blade. The incidence of cytological endometritis in the herd was 22%, and did not differ between the categories analyzed (primiparous versus multiparous) (P>0.05), and the conception rate for first insemination was also similar between primiparous versus multiparous (P>0.05). However, the pregnancy rate at the end of BS was higher in multiparous cows (83.8%) when compared to primiparous (50.0%) cows (P<0.05). The presence or absence of cytological endometritis did not influence the conception rate (P>0.05) nor pregnancy rate at the end of the BS (P>0.05). Therefore, it can be concluded that the use of endometrial cytological cannot be justified as a diagnostic tool in Nelore beef cows.


Subject(s)
Animals , Endometritis/pathology , Neutrophils/cytology , Reproduction/genetics , Uterus/anatomy & histology , Cattle/classification
13.
Neuroscience ; 237: 118-29, 2013 May 01.
Article in English | MEDLINE | ID: mdl-23396085

ABSTRACT

Physical exercise is known to produce beneficial effects to the nervous system. In most cases, brain-derived neurotrophic factor (BDNF) is involved in such effects. However, little is known on the role of BDNF in exercise-related effects on Parkinson's disease (PD). The aim of this study was to investigate the effects of intermittent treadmill exercise-induced behavioral and histological/neurochemical changes in a rat model of unilateral PD induced by striatal injection of 6-hydroxydopamine (6-OHDA), and the role of BDNF in the exercise effects. Adult male Wistar rats were divided into two main groups: (1) injection of K252a (a blocker of BDNF receptors), and (2) without BDNF receptor blockade. These groups were then subdivided into four groups: control (CLT), sedentary (SED, non-exercised with induction of PD), exercised 3×/week during four weeks before and four weeks after the induction of PD (EXB+EXA), and exercised 3×/week during four weeks after the induction of PD (EXA). One month after 6-OHDA injections, the animals were subjected to rotational behavioral test induced by apomorphine and the brains were collected for immunohistochemistry and immunoblotting assays, in which we measured BDNF and tyrosine hydroxylase (TH) in the substantia nigra pars compacta (SNc) and the striatum (caudate-putamen, CPu). Our results showed a significant reduction of rotational asymmetry induced by apomorphine in the exercised parkinsonian rats. BDNF decreased in the SNc of the SED group, and exercise was able to revert that effect. Exercised groups exhibited reduced damage to the dopaminergic system, detected as a decreased drop of TH levels in SNc and CPu. On the other hand, BDNF blockade was capable of substantially reducing TH expression postlesion, implying enhanced dopaminergic cell loss. Our data revealed that physical exercise is capable of reducing the damage induced by 6-OHDA, and that BDNF receptors are involved in that effect.


Subject(s)
Exercise Therapy/methods , Parkinson Disease/rehabilitation , Receptor, trkB/metabolism , Analysis of Variance , Animals , Apomorphine , Carbazoles/pharmacology , Corpus Striatum/drug effects , Corpus Striatum/physiology , Disease Models, Animal , Enzyme Inhibitors/pharmacology , Exercise Test , Gene Expression Regulation/drug effects , Indole Alkaloids/pharmacology , Male , Oxidopamine/toxicity , Parkinson Disease/etiology , Putamen/drug effects , Putamen/physiology , Rats , Rats, Wistar , Stereotyped Behavior/drug effects , Time Factors , Tyrosine 3-Monooxygenase/metabolism
14.
Colloids Surf B Biointerfaces ; 104: 254-61, 2013 Apr 01.
Article in English | MEDLINE | ID: mdl-23333913

ABSTRACT

The development of new polymer-liposome complexes (PLCs) as delivery systems is the key issue of this work. Three main areas are dealt with: polymer synthesis/characterization, liposome formulation/characterization and evaluation of the PLCs uptake by eukaryotic cells. Poly(N,N-dimethylaminoethyl methacrylate) (PDMAEMA) with low molecular weight and narrow polydispersity was synthesized by Atom Transfer Radical Polymerization (ATRP). The polymers were synthesized using two different bromide initiators (cholesteryl-2-bromoisobutyrate and ethyl 2-bromoisobutyrate) as a route to afford PDMAEMA and CHO-PDMAEMA. Both synthesized polymers (PDMAEMA and CHO-PDMAEMA) were incorporated in the preparation of lecithin liposomes (LEC) to obtain PLCs. Three polymer/lipid ratios were investigated: 5, 10 and 20%. Physicochemical characterization of PLCs was carried out by determining the zeta potential, particle size distribution, and the release of fluorescent dyes (carboxyfluorescein CF and calcein) at different temperatures and pHs. The leakage experiments showed that CHO covalently bound to PDMAEMA strongly stabilizes PLCs. The incorporation of 5% CHO-PDMAEMA to LEC (LEC_CHO-PD5) appeared to be the stablest preparation at pH 7.0 and at 37°C. LEC_CHO-PD5 destabilized upon slight changes in pH and temperature, supporting the potential use of CHO-PDMAEMA incorporated to lecithin liposomes (LEC_CHO-PDs) as stimuli-responsive systems. In vitro studies on Raw 264.7 and Caco-2/TC7 cells demonstrated an efficient incorporation of PLCs into the cells. No toxicity of the prepared PLCs was observed according to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. These results substantiate the efficiency of CHO-PDMAEMA incorporated onto LEC to assist for the release of the liposome content in mildly acidic environments, like those found in early endosomes where pH is slightly lower than the physiologic. In summary, the main achievements of this work are: (a) novel synthesis of CHO-PDMAEMA by ATRP, (b) stabilization of LEC by incorporation of CHO-PDMAEMA at neutral pH and destabilization upon slight changes of pH, (c) efficient uptake of LEC_CHO-PDs by phagocytic and non-phagocytic eukaryotic cells.


Subject(s)
Cholesterol/pharmacokinetics , Liposomes/pharmacokinetics , Methacrylates/pharmacokinetics , Nylons/pharmacokinetics , Animals , Caco-2 Cells , Cell Line , Cholesterol/chemistry , Drug Delivery Systems , Fluorescent Dyes/chemistry , Humans , Hydrogen-Ion Concentration , Lecithins/chemistry , Liposomes/chemical synthesis , Liposomes/chemistry , Methacrylates/chemistry , Mice , Molecular Structure , Nylons/chemistry , Particle Size , Polymerization , Surface Properties , Temperature , Tissue Distribution
15.
J Hum Hypertens ; 27(4): 261-4, 2013 Apr.
Article in English | MEDLINE | ID: mdl-22572766

ABSTRACT

The objective of this study was to determine if the maternal serum levels of visfatin in the first trimester of pregnancy are altered in cases that develop preeclampsia (PE) and whether the levels are related to placental perfusion reflected in uterine artery pulsatility index (PI). Serum visfatin and uterine artery PI were measured at 11(+0)-13(+6) weeks in 80 cases that developed PE and 240 unaffected controls. The median visfatin and uterine artery PI multiple of the unaffected median (MoM) in the outcome groups was compared and the significance of the association between visfatin MoM and uterine artery PI MoM, birth weight percentile and gestation at delivery was determined. In the PE group, compared with controls, there was a significantly higher median visfatin MoM (1.35, interquartile range (IQR): 0.69-2.16 vs 1.00, IQR: 0.55-1.96, P=0.027) and uterine artery PI MoM (1.19, IQR: 0.95-1.44 vs 1.03, IQR: 0.83-1.22, P<0.0001). In the PE group, there was no significant association between serum visfatin MoM and uterine artery PI MoM (P=0.589), gestation at delivery (P=0.763) or birth weight percentile (P=0.646). Serum visfatin levels at 11-13 weeks are increased in women who develop PE by a mechanism unrelated to impaired placental perfusion.


Subject(s)
Cytokines/blood , Nicotinamide Phosphoribosyltransferase/blood , Pre-Eclampsia/blood , Adult , Biomarkers/blood , Case-Control Studies , Chi-Square Distribution , Female , Gestational Age , Humans , Placental Circulation , Pre-Eclampsia/diagnostic imaging , Pre-Eclampsia/physiopathology , Pregnancy , Pregnancy Trimester, First/blood , Prospective Studies , Pulsatile Flow , Regional Blood Flow , Ultrasonography, Doppler, Pulsed , Ultrasonography, Prenatal , Up-Regulation , Uterine Artery/physiopathology
16.
Clin Exp Immunol ; 168(3): 291-302, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22519592

ABSTRACT

Type 1 diabetes (T1D) is a chronic autoimmune disease characterized by T cell-mediated destruction of pancreatic ß cells, resulting in insulin deficiency and hyperglycaemia. Recent studies have described that apoptosis impairment during central and peripheral tolerance is involved in T1D pathogenesis. In this study, the apoptosis-related gene expression in T1D patients was evaluated before and after treatment with high-dose immunosuppression followed by autologous haematopoietic stem cell transplantation (HDI-AHSCT). We also correlated gene expression results with clinical response to HDI-AHSCT. We observed a decreased expression of bad, bax and fasL pro-apoptotic genes and an increased expression of a1, bcl-x(L) and cIAP-2 anti-apoptotic genes in patients' peripheral blood mononuclear cells (PBMCs) compared to controls. After HDI-AHSCT, we found an up-regulation of fas and fasL and a down-regulation of anti-apoptotic bcl-x(L) genes expression in post-HDI-AHSCT periods compared to pre-transplantation. Additionally, the levels of bad, bax, bok, fasL, bcl-x(L) and cIAP-1 genes expression were found similar to controls 2 years after HDI-AHSCT. Furthermore, over-expression of pro-apoptotic noxa at 540 days post-HDI-AHSCT correlated positively with insulin-free patients and conversely with glutamic acid decarboxylase autoantibodies (GAD65) autoantibody levels. Taken together, the results suggest that apoptosis-related genes deregulation in patients' PBMCs might be involved in breakdown of immune tolerance and consequently contribute to T1D pathogenesis. Furthermore, HDI-AHSCT modulated the expression of some apoptotic genes towards the levels similar to controls. Possibly, the expression of these apoptotic molecules could be applied as biomarkers of clinical remission of T1D patients treated with HDI-AHSCT therapy.


Subject(s)
Diabetes Mellitus, Type 1/immunology , Diabetes Mellitus, Type 1/surgery , Fas Ligand Protein/genetics , Gene Expression , Hematopoietic Stem Cell Transplantation , Immune Tolerance/genetics , Leukocytes, Mononuclear/drug effects , fas Receptor/genetics , Adolescent , Adult , Apoptosis/genetics , Autoantibodies/metabolism , Down-Regulation , Female , Follow-Up Studies , Glutamate Decarboxylase/immunology , Humans , Immunosuppressive Agents/administration & dosage , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Leukocytes, Mononuclear/pathology , Male , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/immunology , Proto-Oncogene Proteins c-bcl-2/metabolism , Transplantation, Autologous , Up-Regulation , Young Adult , bcl-X Protein/genetics , bcl-X Protein/immunology , bcl-X Protein/metabolism
17.
Bioresour Technol ; 103(1): 459-65, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22029957

ABSTRACT

In this work the parameters of Low Temperature Conversion--LTC were applied in a centrifuged sludge from a sewage treatment plant located in Rio de Janeiro, Brazil. Before the conversion, the sludge was dried and analyzed by TGA to observe its behavior with increasing temperature. The chemical composition of the crude pyrolysis oil was analyzed by FTIR, 1H NMR and GC-MS. The results showed that the oil is a mixture of hydrocarbons, oxygenated and nitrogenated compounds. Using a catalytic treatment it was possible to fractionate the oil where the predominant constituents were hydrocarbons showing that the cracking was effective. An important result was the difference between the calorific value of dry sludge (10 MJ kg(-1)), the pyrolysis oil (36 MJ kg(-1)) and one of the fractions separated by catalytic cracking (40 MJ kg(-1)) when compared with commercial diesel (45 MJ kg(-1)).


Subject(s)
Biotechnology/methods , Oils/chemistry , Sewage/chemistry , Temperature , Water Purification , Catalysis , Chemical Fractionation , Gas Chromatography-Mass Spectrometry , Thermogravimetry
18.
Photodiagnosis Photodyn Ther ; 8(4): 332-6, 2011 Dec.
Article in English | MEDLINE | ID: mdl-22122921

ABSTRACT

This study evaluated the efficacy of PDT in photoinactivation of Candida species using methylene blue (MB) and irradiation with a diode laser (660nm, 40mW). Suspensions of Candida species were obtained containing 10(6)cfu/ml, transferred to 96-holes plates and exposed to 03 doses of laser light (60J/cm(2), 120J/cm(2), 180J/cm(2)) in the presence of MB. Additional suspensions were treated with only the MB, the laser light or with 0.85% saline (control groups). After the treatments, 1µl aliquot of the suspensions was plated in duplicate on SDA. The plates were incubated at 37°C for 24-48h and after this period there was the counting of colonies (cfu/ml). The three evaluated doses determined meaningful inactivation of Candida spp. (p<0.05). The 180J/cm(2) dose was the most effective, inactivating 78% of cfu/ml. At a dose of 180J/cm(2)C. albicans was the most susceptible specie. PDT has demonstrated effectiveness in the inactivation of Candida spp.


Subject(s)
Candida/physiology , Candida/radiation effects , Sterilization/methods , Apoptosis/radiation effects , Cell Survival/radiation effects , Dose-Response Relationship, Radiation , Light , Photochemotherapy/methods , Radiation Dosage
19.
Rev Soc Bras Med Trop ; 34(6): 577-81, 2001.
Article in English | MEDLINE | ID: mdl-11813066

ABSTRACT

"Quantitative Buffy Coat" (QBC) is a direct and fast fluorescent method used for the identification of blood parasites. Since Leishmania chagasi circulates in blood, we decided to test it in American visceral leishmaniasis (AVL). Bone marrow (BM) and peripheral blood (PB) of 49 persons and PB of 31 dogs were analyzed. QBC was positive in BM of 11/11 patients with AVL and in 1/6 patients with other diseases. Amastigotes were identified in PB of 18/22 patients with AVL and in none without AVL. The test was positive in 30 out of the 31 seropositive dogs and in 28/28 dogs with Leishmania identified in other tissues. QBC is a promising method for diagnosis of human AVL, and possibly for the exam of PB of patients with AVL/AIDS, for the control of the cure and for the identification of asymptomatic carriers. Because it is fast and easy to collect and execute, QBC should be evaluated for programs of reservoir control.


Subject(s)
Dog Diseases/diagnosis , Leishmaniasis, Visceral/diagnosis , Leishmaniasis, Visceral/veterinary , Animals , Dogs , Fluorescence , Humans , Parasitology/methods
20.
Cad Saude Publica ; 16(2): 377-84, 2000.
Article in Portuguese | MEDLINE | ID: mdl-10883036

ABSTRACT

This article analyzes knowledge and use of contraceptive methods in women ages 10 to 49 years residing in the southern region of the city of São Paulo in 1992. A total of 1,157 childbearing-age women were studied, focusing on variables that might define them as to: knowledge in the use of contraceptive methods and reasons for choosing a given method. We observed that 86% of the women referred knowledge of some contraceptive method, while the most common was the pill (95.3%), followed by condoms (92.6%). Meanwhile, 14% of the interviewees denied knowledge of any contraceptive method. Of the sexually active women (66.4%), 34.9% reported never having used contraceptive methods. Of those who had, 35.3% used the pill, while 42.9% had resorted to sterilization. Only 5.2% used condoms. Despite the high level of knowledge concerning contraceptive methods, especially oral contraceptives and condoms, we observed limited use of same as compared to the high sterilization rate around the age of 27, thus leaving contraception limited to the pill and female sterilization.


Subject(s)
Contraception Behavior/statistics & numerical data , Contraception/methods , Health Knowledge, Attitudes, Practice , Adolescent , Adult , Age Factors , Brazil , Child , Contraception/statistics & numerical data , Contraceptive Devices, Female/statistics & numerical data , Contraceptives, Oral , Female , Humans , Middle Aged , Sexual Behavior , Sterilization, Tubal/statistics & numerical data
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