RESUMEN
Huntington's disease (HD) is a neurodegenerative disorder caused by a CAG nucleotide expansion, which encodes the amino acid glutamine, in the huntingtin gene. HD is characterized by motor, cognitive, and psychiatric dysfunctions. In a previous study, we showed by qPCR that some genes altered in an HD mouse model were also altered in blood of HD patients. These alterations were mainly with respect to the dynein family. Therefore, this study aimed to investigate whether dynein light chain Tctex type 1 (DYNLT1) is altered in HD patients and if there is a correlation between DYNLT1 gene expression changes and disease progression. We assessed the DYNLT1 gene expression in the blood of 19 HD patients and 20 healthy age-matched controls. Also, in 6 of these patients, we analyzed the DYNLT1 expression at two time points, 3 years apart. The DYNLT1 gene expression in the whole blood of HD patients was significantly downregulated and this difference was widened in later stages. These data suggest that DYNLT1 could emerge as a peripheral prognostic indicator in HD and, also, might be a target for potential intervention in the future.
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Dineínas/genética , Enfermedad de Huntington , Animales , Estudios de Casos y Controles , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Dineínas/sangre , Expresión Génica , Humanos , Proteína Huntingtina/genética , Enfermedad de Huntington/genética , RatonesRESUMEN
Opportunistic infections can cause manifestations that resemble neuropsychiatric systemic lupus erythematosus and they can also trigger lupus flares. Therefore, central nervous system infections as differential diagnosis in neuropsychiatric systemic lupus erythematosus may be difficult, leading to delayed diagnosis and specific treatment. Central nervous system infection in systemic lupus erythematosus is not common but, if left misdiagnosed and not treated promptly, can be fatal. Complementary diagnosis tests are generally non-specific and disappointing. Caution with immunosuppressive drug treatment should be emphasized while an opportunistic infection cannot be ruled out. In this review, we discuss the various types of central nervous system infections reported in systemic lupus erythematosus patients, highlighting the importance of their early recognition in order to improve morbidity and mortality. Prevention with vaccination is a recommended approach.
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Enfermedades del Sistema Nervioso Central/microbiología , Lupus Eritematoso Sistémico/tratamiento farmacológico , Vasculitis por Lupus del Sistema Nervioso Central/diagnóstico , Infecciones Oportunistas/diagnóstico , Diagnóstico Diferencial , Diagnóstico Precoz , Femenino , Humanos , Inmunosupresores/efectos adversos , Inmunosupresores/uso terapéutico , Lupus Eritematoso Sistémico/complicaciones , Masculino , Infecciones Oportunistas/complicaciones , Brote de los SíntomasRESUMEN
OBJECTIVE: To evaluate the immunogenicity of the 23-valent pneumococcal polysaccharide vaccine (PPSV23) in adult systemic lupus erythematosus patients undergoing (IS group) and not undergoing (non-IS group) immunosuppressive treatment. METHODS: In this prospective open-label study from February 2013 to April 2014, 54 patients had blood samples collected immediately before PPSV23 immunization and 4-6 weeks thereafter for the ELISA measurement of IgG antibody levels against seven pneumococcal serotypes. Positive vaccine response for each serotype was defined as a four-fold or greater antibody response over baseline levels or as a post-vaccine anti-pneumococcal IgG level ≥1.3 µg/ml when baseline values were <1.3 µg/ml. Patients should have responded appropriately to ≥70% of the tested serotypes. We also calculated the mean ratio of post- to pre-vaccination anti-pneumococcal IgG levels. RESULTS: Twenty-eight patients were classified into the IS group and 26 into non-IS group. The median dose of prednisone at baseline was ≤5 mg/day in both groups. Serotype-specific vaccine response rates were not significantly different between the groups. Less than 40% of patients responded adequately by both vaccine response criteria, being numerically lower among IS patients. The mean ratio of increase in anti-pneumococcal levels was 6.4 versus 4.7 (p = 0.001) in non-IS and IS groups, respectively. CONCLUSION: The vaccine was poorly immunogenic, especially among adult systemic lupus erythematosus patients under immunosuppressive therapy.
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Inmunosupresores/uso terapéutico , Lupus Eritematoso Sistémico/tratamiento farmacológico , Lupus Eritematoso Sistémico/inmunología , Vacunas Neumococicas/inmunología , Adulto , Femenino , Humanos , Inmunogenicidad Vacunal , Masculino , Persona de Mediana Edad , Estudios Prospectivos , SerogrupoRESUMEN
Huntington's disease (HD) is an autosomal-dominant neurodegenerative disorder caused by a polyglutamine expansion in the amino-terminal region of the huntingtin protein (htt), leading to motor dysfunction, cognitive decline, psychiatric alterations, and death. The metabotropic glutamate receptor 5 (mGluR5) has been implicated in HD and we have recently demonstrated that mGluR5 positive allosteric modulators (PAMs) are neuroprotective in vitro. In the present study we demonstrate that the mGluR5 PAM, CDPPB, is a potent neuroprotective drug, in vitro and in vivo, capable of delaying HD-related symptoms. The HD mouse model, BACHD, exhibits many HD features, including neuronal cell loss, htt aggregates, motor incoordination and memory impairment. However, chronic treatment of BACHD mice with CDPPB 1.5 mg/kg s.c. for 18 weeks increased the activation of cell signaling pathways important for neuronal survival, including increased AKT and ERK1/2 phosphorylation and augmented the BDNF mRNA expression. CDPPB chronic treatment was also able to prevent the neuronal cell loss that takes place in the striatum of BACHD mice and decrease htt aggregate formation. Moreover, CDPPB chronic treatment was efficient to partially ameliorate motor incoordination and to rescue the memory deficit exhibited by BACHD mice. Importantly, no toxic effects or stereotypical behavior were observed upon CDPPB chronic treatment. Thus, CDPPB is a potential drug to treat HD, preventing neuronal cell loss and htt aggregate formation and delaying HD symptoms.
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Benzamidas/uso terapéutico , Enfermedad de Huntington/tratamiento farmacológico , Enfermedad de Huntington/patología , Enfermedad de Huntington/fisiopatología , Neuronas/efectos de los fármacos , Pirazoles/uso terapéutico , Factores de Edad , Animales , Muerte Celular/efectos de los fármacos , Células Cultivadas , Cuerpo Estriado/citología , Modelos Animales de Enfermedad , Embrión de Mamíferos , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Ácido Glutámico/farmacología , Humanos , Proteína Huntingtina , Enfermedad de Huntington/genética , Ratones , Ratones Transgénicos , Mitocondrias/patología , Actividad Motora/efectos de los fármacos , Actividad Motora/genética , Proteínas del Tejido Nervioso/metabolismo , Neuronas/patología , Reconocimiento en Psicología/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Sinapsis/patología , Sinapsis/ultraestructuraRESUMEN
Introduction. The positioning ofγray interactions in positron emission tomography (PET) detectors is commonly made through the evaluation of the Anger logic flood histograms. machine learning techniques, leveraging features extracted from signal waveform, have demonstrated successful applications in addressing various challenges in PET instrumentation.Aim. This paper evaluates the use of artificial neural networks (NN) forγray interaction positioning in pixelated scintillators coupled to a multiplexed array of silicon photomultipliers (SiPM).Methods. An array of 16 Cerium doped Lutetium-based (LYSO) crystal pixels (cross-section 2 × 2 mm2) coupled to 16 SiPM (S13360-1350) were used for the experimental setup. Data from each of the 16 LYSO pixels was recorded, a total of 160000 events. The detectors were irradiated by 511 keV annihilationγrays from a Sodium-22 (22Na) source. Another LYSO crystal was used for electronic collimation. Features extracted from the signal waveform were used to train the model. Two models were tested: i) single multiple-class neural network (mcNN), with 16 possible outputs followed by a softmax and ii) 16 binary classification neural networks (bNN), each one specialized in identifying events occurred in each position.Results. Both NN models showed a mean positioning accuracy above 85% on the evaluation dataset, although the mcNN is faster to train.DiscussionThe method's accuracy is affected by the introduction of misclassified events that interacted in the neighbour's crystals and were misclassified during the dataset acquisition. Electronic collimation reduces this effect, however results could be improved using a more complex acquisition setup, such as a light-sharing configuration.ConclusionsThe methods comparison showed that mcNN and bNN can surpass the Anger logic, showing the feasibility of using these models in positioning procedures of future multiplexed detector systems in a linear configuration.
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Rayos gamma , Redes Neurales de la Computación , Tomografía de Emisión de Positrones , Tomografía de Emisión de Positrones/métodos , Conteo por Cintilación/instrumentación , Conteo por Cintilación/métodos , Lutecio/química , Cerio/química , Silicio/química , Algoritmos , Diseño de EquipoRESUMEN
OBJECTIVE: To determine the epidemiological profile and outcome of patients with lupus nephritis (LN) undergoing renal transplantation. METHODS: The archival records of 50 patients with LN and end-stage renal disease (ESRD) treated by kidney transplantation from March 1992 to December 2010 were reviewed. All patients met the American College of Rheumatology criteria for systemic lupus erythematosus (SLE). RESULTS: Fourteen patients were included in the study. The majority were women (85.7%) and non-Caucasian (85.7%); the mean age at diagnosis of SLE and LN was 24 ± 8 and 25 ± 8 years, respectively. Renal biopsy was performed in 12 patients, with 75% of them showing proliferative lesions (class III and IV according to the World Health Organization and International Society of Nephrology/Renal Pathology Society classification). Thirteen patients (93%) underwent intermittent hemodialysis or peritoneal dialysis before transplantation. The median time between the start of dialysis and transplantation was 30 months (range 3-103 months); 67% of the procedures involved deceased donors and 33% involved living-related donors. The graft survival rates were 93.3%, 90.9%, and 85.7% at 1, 5 and 10 years, respectively. Post-transplant immunosuppressive agents were mycophenolate mofetil (84%), azathioprine (17%), tacrolimus (25%), sirolimus (58%) and cyclosporine (8%). Eight episodes of acute rejection were noted in six patients. There was a graft loss due to renal vein thrombosis in the one patient with secondary antiphospholipid syndrome. The mean SLICC by the time of kidney transplantation was 5 ± 2. In total, 13 patients (92.8%) developed at least one infectious event during the follow-up, with one dying in the immediate post-transplant period because of sepsis. Two patients (14%) had a lupus flare. There was no clinical or histological evidence of LN recurrence. CONCLUSION: LN is the major cause of morbidity in SLE, with progression to ESRD in 10-22% of cases. Despite concerns about LN recurrence after renal transplantation, the data obtained in our sample indicate this procedure as a safe alternative therapy for ESRD in this population.
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Fallo Renal Crónico/cirugía , Trasplante de Riñón/métodos , Nefritis Lúpica/cirugía , Adulto , Brasil/epidemiología , Progresión de la Enfermedad , Femenino , Estudios de Seguimiento , Rechazo de Injerto/epidemiología , Supervivencia de Injerto , Humanos , Inmunosupresores/uso terapéutico , Fallo Renal Crónico/etiología , Nefritis Lúpica/complicaciones , Nefritis Lúpica/epidemiología , Masculino , Diálisis Peritoneal/métodos , Recurrencia , Diálisis Renal/métodos , Factores de Tiempo , Resultado del Tratamiento , Adulto JovenRESUMEN
The experimental outcomes of small-animal positron emission tomography (PET) imaging with 18F-labelled fluorodeoxyglucose (18F-FDG) can be particularly compromised by animal preparation and care. Several works intend to improve research reporting and amplify the quality and reliability of published research. Though these works provide valuable information to plan and conduct animal studies, manuscripts describe different methodologies-standardization does not exist. Consequently, the variation in details reported can explain the difference in the experimental results found in the literature. Additionally, the resources and guidelines defining protocols for small-animal imaging are scarce, making it difficult for researchers to obtain and compare accurate and reproducible data. Considering the selection of suitable procedures key to ensure animal welfare and research improvement, this paper aims to prepare the way for a future guideline on mice preparation and care for PET imaging with 18F-FDG. For this purpose, a global standard protocol was created based on recommendations and good practices described in relevant literature.
RESUMEN
Human papillomavirus (HPV) is the etiological agent of cervical cancer, the second most prevalent neoplasia among women. Although it has been proven that systemic lupus erythematosus (SLE) patients have higher frequency of cervical dysplasia, few studies have focused on HPV prevalence among them. This study aimed to investigate HPV prevalence among SLE patients and to evaluate associated risk factors, including the use of immunosuppressors (IM). Total DNA extracted from cervical samples of 173 SLE patients and 217 women (control group) submitted to routine cervical cytopathology was used as template in polymerase chain reaction (PCR)-based assays for detection of HPV DNA. HPV genotyping was performed by type-specific PCR, PCR-RFLP and/or DNA sequencing. Statistical methods included univariate analysis and logistic regression. Despite presenting significantly fewer HPV risk factors, SLE patients were found to have a threefold increase in HPV infection, mostly genotypes 53, 58, 45, 66, 6, 84, 83, 61, as compared with controls, who presented types 6, 18 and 61 more frequently. The higher rate of HPV infection was associated with immunosuppressive therapy. This study provides evidence that SLE patients have a high prevalence of HPV infection, which is even higher with the use of IM, a condition that might necessitate a more frequent cervical cancer screening program for these women.
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Inmunosupresores/efectos adversos , Lupus Eritematoso Sistémico/complicaciones , Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/etiología , Adolescente , Adulto , Anciano , Secuencia de Bases , Estudios de Casos y Controles , Estudios Transversales , ADN Viral/análisis , Femenino , Genotipo , Humanos , Inmunosupresores/uso terapéutico , Modelos Logísticos , Lupus Eritematoso Sistémico/tratamiento farmacológico , Lupus Eritematoso Sistémico/epidemiología , Persona de Mediana Edad , Papillomaviridae/genética , Infecciones por Papillomavirus/epidemiología , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Prevalencia , Estudios Prospectivos , Factores de Riesgo , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/epidemiología , Neoplasias del Cuello Uterino/virología , Frotis Vaginal , Adulto JovenRESUMEN
It has been suggested that overexpression of neuronal Ca2+ sensor-1 (NCS-1) protein is implicated in the pathophysiology of neurodisorders such as schizophrenia, bipolar disturbance and X-linked mental retardation. The mechanism by which NCS-1 would be involved in the causes and/or consequences of these neurodisorders is still far from elucidation. Independent evidence has pointed NCS-1 as a key regulator of synaptic efficacy by altering the expression and activity of voltage-gated channels, inhibiting internalization of dopaminergic receptors, and altering phosphoinositide metabolism. In this study, we examined the possible participation of NCS-1 protein in signal transmission dependent on muscarinic receptor activation, using PC12 cells stably expressing NCS-1 (PC12-NCS-1). Carbachol (CCH; 300 microM) was able to evoke glutamate release more efficiently from PC12-NCS-1 (15.3+/-1.0nmol/mg of protein) than wild type cells (PC12-wt; 8.3+/-0.9nmol/mg of protein). This increase of glutamate release induced by CCH was independent on extracellular Ca2+ influx. Additionally, a larger increase of cytoplasmic levels of InsP3 (663.0+/-63.0 and 310.0+/-39.0% of fluorescence in A.U.) and [Ca2+]i (766.4+/-40.0 and 687.8+/-37.1nmol/L) was observed after CCH stimulus of PC12-NCS-1 compared with PC12-wt. Clearly distinction between intracellular Ca2+ dynamics was also observed in PC12-NCS-1 and PC12-wt. A larger increase followed by fast decay of [Ca2+]i was observed in PC12-NCS-1. A plateau with a delayed decay of [Ca2+]i was characteristic of PC12-wt [Ca2+]i response. Both enhancement of InsP3 production and glutamate release observed in PC12-NCS-1 were blocked by atropine (10 microM). Together, our data show that overexpression of NCS-1 in PC12 cells induces an enhancement of intracellular second messenger and transmitter release dependent on CCH response, suggesting that muscarinic signaling is "up-regulated" in this cell model.
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Ácido Glutámico/metabolismo , Proteínas Sensoras del Calcio Neuronal/fisiología , Neuropéptidos/fisiología , Receptores Muscarínicos/metabolismo , Transducción de Señal/fisiología , Animales , Calcio/metabolismo , Calcio/farmacología , Carbacol/farmacología , Quelantes/farmacología , Agonistas Colinérgicos/farmacología , Ácido Egtácico/análogos & derivados , Ácido Egtácico/farmacología , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Microscopía Fluorescente , Proteínas Sensoras del Calcio Neuronal/genética , Proteínas Sensoras del Calcio Neuronal/metabolismo , Neuropéptidos/genética , Neuropéptidos/metabolismo , Células PC12 , Transporte de Proteínas/efectos de los fármacos , Ratas , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , TransfecciónRESUMEN
Systemic lupus erythematous (SLE) is an autoimmune disease that presents an increased susceptibility to infections which may trigger reactivation. Disease flares have been mostly associated with parvovirus B19, cytomegalovirus, EBV and Mycobacterium tuberculosis infections, but it is probable that many other agents may also induce innate and adaptive immune system stimulation including the production of autoantibodies as ANA, anti nDNA and anti-ß2-GPI mainly in lepromatous leprosy. Mycobacterium leprae not only may determine symptoms that mimic lupus flares, including autoantibodies production, but could also act as a trigger for lupus reactivation; however, its association is still not fully explored. As demonstrated for tuberculosis, it is quite possible that molecular mimicry may also be involved in the interface of these two diseases. Some studies reported shared epitopes among idiotypes derived from 8E7 and TH9 lepromatous antibodies and those obtained from SLE patients, and it could partially explain the triggering phenomenon of SLE caused by M. leprae. We report and discuss three Brazilian patients whose disease was inactive and presented disease flares concurrently with the diagnosis of leprosy.
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Lepra/complicaciones , Lupus Eritematoso Sistémico/complicaciones , Adulto , Biopsia , Femenino , Humanos , Inmunosupresores/uso terapéutico , Lepra/diagnóstico , Lepra/tratamiento farmacológico , Lupus Eritematoso Sistémico/tratamiento farmacológico , Masculino , Resultado del TratamientoRESUMEN
OBJECTIVES: To assess the bioequivalence of 2 formulations of carbamazepine and to develop and validate limited sampling strategy (LSS) models for estimating the area under the plasma concentration-time curve (AUC0-infinity) and the peak plasma concentration (Cmax) of carbamazepine. METHODS: Twenty-four (12 men, 12 women) healthy volunteers received single oral doses (400 mg) of carbamazepine, as reference and test conventional-release formulations, in a standard 2-sequence, 2-period crossover design. Bioequivalence assessment was based on the individual ratios of log-transformed values of AUC0-infinity and Cmax LSS modeling was developed in a training set of 12 randomly assigned volunteers and was validated on the other 12 subjects (validation set). RESULTS: Carbamazepine AUC0-infinity and Cmax can be accurately predicted (R2 = 0.89 - 0.95, precision = 2.6 - 7.2%) by single-point (72 h) and 2-point LSS models (6, 32 h), respectively. Bioequivalence assessments based on LSS-derived AUC0-infinity and Cmax provided results similar to those obtained using all the concentration-in-plasma data points, and indicated that the 2 formulations are bioequivalent. CONCLUSION: One-and 2-point LSS models provided accurate estimates of carbamazepine's AUC0-infinity and Cmax, and allowed correct assessment of bioequivalence between the formulations studied.
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Anticonvulsivantes/farmacocinética , Carbamazepina/farmacocinética , Adulto , Anticonvulsivantes/sangre , Área Bajo la Curva , Carbamazepina/sangre , Cromatografía Líquida de Alta Presión , Femenino , Semivida , Humanos , Masculino , Tasa de Depuración Metabólica , Equivalencia TerapéuticaRESUMEN
Bioanalytical data from a bioequivalence study were used to develop limited-sampling strategy (LSS) models for estimating the area under the plasma concentration versus time curve (AUC) and the peak plasma concentration (Cmax) of 4-methylaminoantipyrine (MAA), an active metabolite of dipyrone. Twelve healthy adult male volunteers received single 600 mg oral doses of dipyrone in two formulations at a 7-day interval in a randomized, crossover protocol. Plasma concentrations of MAA (N = 336), measured by HPLC, were used to develop LSS models. Linear regression analysis and a "jack-knife" validation procedure revealed that the AUC(0-infinity) and the Cmax of MAA can be accurately predicted (R2>0.95, bias <1.5%, precision between 3.1 and 8.3%) by LSS models based on two sampling times. Validation tests indicate that the most informative 2-point LSS models developed for one formulation provide good estimates (R2>0.85) of the AUC(0-infinity) or Cmax for the other formulation. LSS models based on three sampling points (1.5, 4 and 24 h), but using different coefficients for AUC(0-infinity) and Cmax, predicted the individual values of both parameters for the enrolled volunteers (R2>0.88, bias = -0.65 and -0.37%, precision = 4.3 and 7.4%) as well as for plasma concentration data sets generated by simulation (R2>0.88, bias = -1.9 and 8.5%, precision = 5.2 and 8.7%). Bioequivalence assessment of the dipyrone formulations based on the 90% confidence interval of log-transformed AUC(0-infinity) and Cmax provided similar results when either the best-estimated or the LSS-derived metrics were used.
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Dipirona/análogos & derivados , Dipirona/farmacocinética , Pirazolonas , Adulto , Área Bajo la Curva , Intervalos de Confianza , Estudios Cruzados , Dipirona/sangre , Humanos , Masculino , Equivalencia TerapéuticaRESUMEN
BACKGROUND AND PURPOSE: Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by a polyglutamine expansion in the huntingtin protein. We have previously demonstrated that the cell signalling of the metabotropic glutamate receptor 5 (mGluR5) is altered in a mouse model of HD. Although mGluR5-dependent protective pathways are more activated in HD neurons, intracellular Ca²âº release is also more pronounced, which could contribute to excitotoxicity. In the present study, we aim to investigate whether mGluR5 positive allosteric modulators (PAMs) could activate protective pathways without triggering high levels of Ca²âº release and be neuroprotective in HD. EXPERIMENTAL APPROACH: We performed a neuronal cell death assay to determine which drugs are neuroprotective, Western blot and Ca²âº release experiments to investigate the molecular mechanisms involved in this neuroprotection, and object recognition task to determine whether the tested drugs could ameliorate HD memory deficit. KEY RESULTS: We find that mGluR5 PAMs can protect striatal neurons from the excitotoxic neuronal cell death promoted by elevated concentrations of glutamate and NMDA. mGluR5 PAMs are capable of activating Akt without triggering increased intracellular Ca²âº concentration ([Ca²âº]i ); and Akt blockage leads to loss of PAM-mediated neuroprotection. Importantly, PAMs' potential as drugs that may be used to treat neurodegenerative diseases is highlighted by the neuroprotection exerted by mGluR5 PAMs on striatal neurons from a mouse model of HD, BACHD. Moreover, mGluR5 PAMs can activate neuroprotective pathways more robustly in BACHD mice and ameliorate HD memory deficit. CONCLUSIONS AND IMPLICATIONS: mGluR5 PAMs are potential drugs that may be used to treat neurodegenerative diseases, especially HD.
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Enfermedad de Huntington/tratamiento farmacológico , Trastornos de la Memoria/prevención & control , Proteínas del Tejido Nervioso/agonistas , Fármacos Neuroprotectores/uso terapéutico , Nootrópicos/uso terapéutico , Receptor del Glutamato Metabotropico 5/agonistas , Regulación Alostérica/efectos de los fármacos , Animales , Conducta Animal/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Células Cultivadas , Cuerpo Estriado/citología , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/metabolismo , Embrión de Mamíferos/citología , Activación Enzimática/efectos de los fármacos , Proteína Huntingtina , Enfermedad de Huntington/metabolismo , Enfermedad de Huntington/fisiopatología , Trastornos de la Memoria/etiología , Ratones , Ratones Endogámicos , Ratones Transgénicos , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Neuronas/citología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Fármacos Neuroprotectores/efectos adversos , Fármacos Neuroprotectores/farmacología , Nootrópicos/efectos adversos , Nootrópicos/farmacología , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fosforilación/efectos de los fármacos , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/agonistas , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptor del Glutamato Metabotropico 5/metabolismoRESUMEN
Cholinergic neurons rely on the sodium-dependent choline transporter CHT to provide choline for synthesis of acetylcholine. CHT cycles between cell surface and subcellular organelles, but little is known about regulation of this trafficking. We hypothesized that activation of protein kinase C with phorbol ester modulates choline uptake by altering the rate of CHT internalization from or delivery to the plasma membrane. Using SH-SY5Y cells that stably express rat CHT, we found that exposure of cells to phorbol ester for 2 or 5 min significantly increased choline uptake, whereas longer treatment had no effect. Kinetic analysis revealed that 5 min phorbol ester treatment significantly enhanced V(max) of choline uptake, but had no effect on K(m) for solute binding. Cell-surface biotinylation assays showed that plasma membrane levels of CHT protein were enhanced following 5 min phorbol ester treatment; this was blocked by protein kinase C inhibitor bisindolylmaleimide-I. Moreover, CHT internalization was decreased and delivery of CHT to plasma membrane was increased by phorbol ester. Our results suggest that treatment of neural cells with the protein kinase C activator phorbol ester rapidly and transiently increases cell surface CHT levels and this corresponds with enhanced choline uptake activity which may play an important role in replenishing acetylcholine stores following its release by depolarization.
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Encéfalo/metabolismo , Proteínas del Tejido Nervioso/efectos de los fármacos , Neuronas/efectos de los fármacos , Proteínas de Transporte de Neurotransmisores en la Membrana Plasmática/efectos de los fármacos , Proteína Quinasa C/efectos de los fármacos , Acetato de Tetradecanoilforbol/análogos & derivados , Acetilcolina/biosíntesis , Animales , Línea Celular Tumoral , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Endocitosis/efectos de los fármacos , Endocitosis/fisiología , Activación Enzimática/efectos de los fármacos , Activación Enzimática/fisiología , Inhibidores Enzimáticos/farmacología , Cinética , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Proteínas del Tejido Nervioso/metabolismo , Neuronas/metabolismo , Fosforilación/efectos de los fármacos , Fosforilación/fisiología , Proteínas de Transporte de Neurotransmisores en la Membrana Plasmática/metabolismo , Proteína Quinasa C/metabolismo , Transporte de Proteínas/efectos de los fármacos , Transporte de Proteínas/fisiología , Ratas , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Systemic lupus erythematosus (SLE) is an autoimmune disease that frequently requires treatment with high doses of corticosteroids and immunosuppressive drugs. Primary defects in the innate immunity also contribute to an increased susceptibility to infections. Patients with SLE are at an increased risk for infections with several pathogens, among them Mycobacterium tuberculosis, which is a significant cause of morbidity and mortality, especially in endemic regions. TB infection requires awareness for several reasons: first, TB infection thrives under conditions of immunosuppression, may it be secondary to the disease itself or its treatment. Second, shared antigens by mycobacteria and autoantigens have been described, which may be targets for autoantibodies. We present four Brazilian patients, in whom a diagnosis of tuberculosis was determined during or following persistent flares of their disease. The association of SLE and TB is discussed, as well as different aspects of the tuberculosis infection in this selected population, and its possible role in the course of SLE.
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Lupus Eritematoso Sistémico/complicaciones , Tuberculosis/complicaciones , Adulto , Antituberculosos/uso terapéutico , Femenino , Humanos , Inmunosupresores/uso terapéutico , Lupus Eritematoso Sistémico/tratamiento farmacológico , Lupus Eritematoso Sistémico/fisiopatología , Persona de Mediana Edad , Tuberculosis/tratamiento farmacológico , Tuberculosis/fisiopatologíaRESUMEN
Formyl-Met-Leu-Phe (fMLP) is a potent chemoattractant molecule released from both bacteria and damaged mitochondria that activates fMLP receptors (FPR) leading to neutrophil chemotaxis, degranulation and superoxide production. A common missense single nucleotide polymorphism in the human FPR1 gene at nucleotide c.32C>T results in the amino-acid substitution, p.I11T, in the FPR1 extracellular amino-terminus. The minor (c.32T) allele frequencies were 0.25, 0.27, 0.25, 0.15 and 0.14 in healthy Caucasian, African, East Indian, Chinese and Native Canadian individuals, respectively. In subjects homozygous for the p.T11 allele, we find elevated serum concentrations of C-reactive protein, increased absolute counts of blood leukocytes and neutrophils, and erythrocyte sedimentation rates. When expressed in HEK 293 and RBL-2H3 cells a substantial proportion of FPR1 p.I11T variant is retained intracellularly and agonist-independent internalization of the FPR1 p.I11T variant, but not the wild-type FPR1, is constitutively associated with beta-arrestin2-GFP in vesicles. Moreover, basal N-acetyl-D-glucosaminidase release is increased in primary neutrophils isolated from subjects either heterozygous or homozygous for the FPR1 p.T11 allele. Taken together, the data suggest an increased receptor activity and phenotypic expression of increased inflammatory indices in subjects with the p.T11 allele.
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Arrestinas/fisiología , Proteína C-Reactiva/análisis , Inflamación/etiología , Mutación Missense , Receptores de Formil Péptido/genética , Degranulación de la Célula , Línea Celular , Citoesqueleto/metabolismo , Humanos , Neutrófilos/fisiología , Polimorfismo de Nucleótido Simple , beta-ArrestinasRESUMEN
Amoxicillin plasma concentrations (n = 1,152) obtained from 48 healthy subjects in two bioequivalence studies were used to develop limited-sampling strategy (LSS) models for estimating the area under the concentration-time curve (AUC), the maximum concentration of drug in plasma (C(max)), and the time interval of concentration above MIC susceptibility breakpoints in plasma (T>MIC). Each subject received 500-mg amoxicillin, as reference and test capsules or suspensions, and plasma concentrations were measured by a validated microbiological assay. Linear regression analysis and a "jack-knife" procedure revealed that three-point LSS models accurately estimated (R(2), 0.92; precision, <5.8%) the AUC from 0 h to infinity (AUC(0-infinity)) of amoxicillin for the four formulations tested. Validation tests indicated that a three-point LSS model (1, 2, and 5 h) developed for the reference capsule formulation predicts the following accurately (R(2), 0.94 to 0.99): (i) the individual AUC(0-infinity) for the test capsule formulation in the same subjects, (ii) the individual AUC(0-infinity) for both reference and test suspensions in 24 other subjects, and (iii) the average AUC(0-infinity) following single oral doses (250 to 1,000 mg) of various amoxicillin formulations in 11 previously published studies. A linear regression equation was derived, using the same sampling time points of the LSS model for the AUC(0-infinity), but using different coefficients and intercept, for estimating C(max). Bioequivalence assessments based on LSS-derived AUC(0-infinity)'s and C(max)'s provided results similar to those obtained using the original values for these parameters. Finally, two-point LSS models (R(2) = 0.86 to 0.95) were developed for T>MICs of 0.25 or 2.0 microg/ml, which are representative of microorganisms susceptible and resistant to amoxicillin.
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Amoxicilina/farmacocinética , Penicilinas/farmacocinética , Adulto , Área Bajo la Curva , Estudios Cruzados , Método Doble Ciego , Femenino , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Micrococcus luteus/efectos de los fármacos , Valor Predictivo de las Pruebas , Reproducibilidad de los Resultados , Equivalencia TerapéuticaRESUMEN
The detection of high circulating levels of calcitonin is a most valuable procedure to diagnose advanced cases of medullary carcinoma of the thyroid. However, the diagnosis of a primitive tumor in the early stages of development (as in cases of the familial form of the disease) or of a metastasis following ablation of the tumor is more difficult. In the latter cases, the levels of circulating calcitonin may be within normal limits, and for diagnosis one must then resort to tests to stimulate the secretion of calcitonin. We are reporting from our personal experiences the advantages, disadvantages and inconveniences of the three most well-known tests. All patients responded positively to administration of calcium or pentagastrin. The alcohol test, however, produced inconsistent results.
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Calcitonina/metabolismo , Carcinoma/diagnóstico , Neoplasias de la Tiroides/diagnóstico , Adulto , Calcio , Ensayos Clínicos como Asunto , Etanol , Femenino , Humanos , Persona de Mediana Edad , Metástasis de la Neoplasia , Pentagastrina , Radioinmunoensayo , Tasa de Secreción , Estimulación QuímicaRESUMEN
Hemolytic uremic syndrome (HUS) is a rare condition which most frequently follows gastrointestinal or respiratory infection episodes in young children, but it can also occur in other settings such as the postpartum period and during use of drugs such as oral contraconceptives, immunosuppressors, and antineoplastics. In early pregnancy, however, its frequency is thought to be very low. The authors report a case of a 30-year-old woman who developed HUS early in her first pregnancy. She had persistent aqueous diarrhea from the beginning of the pregnancy. At the 21st week she developed hypertension which in 2 weeks was followed by seizures, oliguria, and acute pulmonary edema despite intensive medical efforts to control her blood pressure. Surgical intervention for fetal delivery was performed. The patient was initially kept on continuous hemodialysis (CVVHD) followed by an alternate-day conventional hemodialysis schedule. A peripheral blood analysis showed a microangiopathic hemolytic anemia with thrombocytopenia; blood coagulation tests were completely normal. A brain CT scan and an abdominal MRI showed no major abnormalities. HUS was confirmed by a percutaneal kidney biopsy, performed at the 21st day of anuria. Techniques for identification of verotoxin-producing E. coli were not available. Renal function did not recover and the patient has been undergoing regular maintenance hemodialysis for a year.
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Síndrome Hemolítico-Urémico/diagnóstico , Complicaciones del Embarazo/diagnóstico , Adulto , Femenino , Síndrome Hemolítico-Urémico/terapia , Humanos , Embarazo , Complicaciones del Embarazo/terapia , Complicaciones Hematológicas del Embarazo/diagnóstico , Complicaciones Hematológicas del Embarazo/terapia , Diálisis RenalRESUMEN
Synthesis of acetylcholine depends on the plasma membrane uptake of choline by a high affinity choline transporter (CHT1). Choline uptake is regulated by nerve impulses and trafficking of an intracellular pool of CHT1 to the plasma membrane may be important for this regulation. We have generated a hemagglutinin (HA) epitope tagged CHT1 to investigate the organelles involved with intracellular trafficking of this protein. Expression of CHT1-HA in HEK 293 cells establishes Na+-dependent, hemicholinium-3 sensitive high-affinity choline transport activity. Confocal microscopy reveals that CHT1-HA is found predominantly in intracellular organelles in three different cell lines. Importantly, CHT1-HA seems to be continuously cycling between the plasma membrane and endocytic organelles via a constitutive clathrin-mediated endocytic pathway. In a neuronal cell line, CHT1-HA colocalizes with the early endocytic marker green fluorescent protein (GFP)-Rab 5 and with two markers of synaptic-like vesicles, VAMP-myc and GFP-VAChT, suggesting that in cultured cells CHT1 is present mainly in organelles of endocytic origin. Subcellular fractionation and immunoisolation of organelles from rat brain indicate that CHT1 is present in synaptic vesicles. We propose that intracellular CHT1 can be recruited during stimulation to increase choline uptake in nerve terminals.