RESUMEN
OBJECTIVE: The objective of this study is to assess the autonomic nerve heart rate regulation system at rest and its immediate response to paced breathing among patients with complex regional pain syndrome (CRPS) as compared with age-matched healthy controls. DESIGN: Quasiexperimental. SETTING: Outpatient clinic. SUBJECTS: Ten patients with CRPS and 10 age- and sex-matched controls. METHODS: Participants underwent Holter ECG (NorthEast Monitoring, Inc., Maynard, MA, USA) recording during rest and biofeedback-paced breathing session. Heart rate variability (HRV), time, and frequency measures were assessed. RESULTS: HRV and time domain values were significantly lower at rest among patients with CRPS as compared with controls. A significant association was noted between pain rank and HRV frequency measures at rest and during paced breathing; although both groups reduced breathing rate significantly during paced breathing, HRV time domain parameters increased only among the control group. CONCLUSIONS: The increased heart rate and decreased HRV at rest in patients with CRPS suggest a general autonomic imbalance. The inability of the patients to increase HRV time domain values during paced breathing may suggest that these patients have sustained stress response with minimal changeability in response to slow-paced breathing stimuli.
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Contencion de la Respiración , Síndromes de Dolor Regional Complejo/fisiopatología , Sistema de Conducción Cardíaco/fisiopatología , Frecuencia Cardíaca/fisiología , Descanso/fisiología , Adolescente , Adulto , Antropometría , Estudios de Casos y Controles , Evaluación de la Discapacidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
BACKGROUND: Stroke is the leading cause of adult disability, with walking impairment being a devastating indicator of chronic post-stroke hemiparesis. Limited resources exist for individual treatments; therefore, the delivery of safe group exercise therapy is highly desired. OBJECTIVE: To examine whether the application of group-based motor imagery practice to community-dwelling individuals with chronic hemiparesis improves gait. METHODS: Sixteen individuals with chronic hemiparesis from two community centers participated in the study, with eight from each center. Four participants in each center received five weeks of the experimental intervention, consisting of group-based motor imagery exercises of gait tasks, followed by five weeks of control treatment of motor imagery exercises for the affected upper extremity. Four other subjects in each center received the same treatments in reverse order. Pre- and post intervention measurements included clinical and biomechanical gait parameters. RESULTS: Comparisons within (pre- vs. post) and between treatments (experimental vs. control) indicated no significant change in any gait variable. Nevertheless, the verbal reports of most participants alluded to satisfaction with the experimental intervention and to an increase in self-confidence. CONCLUSIONS: Despite the lack of evidence for the effectiveness of group-based motor imagery practice in improving gait among individuals with chronic hemiparesis, the contrast between the measured outcomes and the positive verbal reports merits further inquiry.
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Trastornos Neurológicos de la Marcha/rehabilitación , Paresia/rehabilitación , Rehabilitación de Accidente Cerebrovascular , Anciano , Ejercicio Físico , Femenino , Marcha , Trastornos Neurológicos de la Marcha/etiología , Humanos , Imágenes en Psicoterapia , Masculino , Persona de Mediana Edad , Destreza Motora , Paresia/etiología , Satisfacción del Paciente , Modalidades de Fisioterapia , Proyectos Piloto , Recuperación de la Función , Accidente Cerebrovascular/complicaciones , Extremidad SuperiorRESUMEN
BACKGROUND AND PURPOSE: This study assessed the potential therapeutic benefi t of using HandTutor™ in combination with traditional rehabilitation in a post-stroke sub-acute population. The study compares an experimental group receiving traditional therapy combined with HandTutorTM treatment, against a control group receiving only traditional therapy. METHOD: An assessor-blinded, randomized controlled pilot trial, was conducted in the Reuth rehabilitation unit in Israel. Thirty-one stroke patients in the sub-acute phase, were randomly assigned to one of the two groups (experimental or control) in sets of three. The experimental group (n = 16) underwent a hand rehabilitation programme using the HandTutorTM combined with traditional therapy. The control group (n = 15) received only traditional therapy. The treatment schedules for both groups were of similar duration and frequency. Improvements were evaluated using three indicators: 1) The Brunnström-Fugl-Meyer (FM) test, 2) the Box and Blocks (B&B) test and 3) improvement parameters as determined by the HandTutorTM software. RESULTS: Following 15 consecutive treatment sessions, a signifi cant improvement was observed within the experimental group (95% confi dence intervals) compared with the control group: B&B p = 0.015; FM p = 0.041, HandTutor™ performance accuracy on x axis and performance accuracy on y axis p < 0.0003. CONCLUSION: The results from this pilot study support further investigation of the use of the HandTutorTM in combination with traditional occupational therapy and physiotherapy during post stroke hand function rehabilitation.
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Fuerza de la Mano/fisiología , Mano/fisiología , Modalidades de Fisioterapia/instrumentación , Rehabilitación/instrumentación , Rehabilitación/métodos , Rehabilitación de Accidente Cerebrovascular , Anciano , Terapia por Ejercicio/métodos , Femenino , Humanos , Israel , Masculino , Persona de Mediana Edad , Terapia Ocupacional/métodos , Especialidad de Fisioterapia/métodos , Proyectos Piloto , Método Simple Ciego , Programas Informáticos , Accidente Cerebrovascular/fisiopatología , Análisis y Desempeño de Tareas , Resultado del TratamientoRESUMEN
Utilizing the Citrobacter rodentium-induced transmissible murine colonic hyperplasia (TMCH) model, we measured hyperplasia and NF-κB activation during progression (days 6 and 12 post-infection) and regression (days 20-34 post-infection) phases of TMCH. NF-κB activity increased at progression in conjunction with bacterial attachment and translocation to the colonic crypts and decreased 40% by day 20. NF-κB activity at days 27 and 34, however, remained 2-3-fold higher than uninfected control. Expression of the downstream target gene CXCL-1/KC in the crypts correlated with NF-κB activation kinetics. Phosphorylation of cellular IκBα kinase (IKK)α/ß (Ser(176/180)) was elevated during progression and regression of TMCH. Phosphorylation (Ser(32/36)) and degradation of IκBα, however, contributed to NF-κB activation only from days 6 to 20 but not at later time points. Phosphorylation of MEK1/2 (Ser(217/221)), ERK1/2 (Thr(202)/Tyr(204)), and p38 (Thr(180)/Tyr(182)) paralleled IKKα/ß kinetics at days 6 and 12 without declining with regressing hyperplasia. siRNAs to MEK, ERK, and p38 significantly blocked NF-κB activity in vitro, whereas MEK1/2-inhibitor (PD98059) also blocked increases in MEK1/2, ERK1/2, and IKKα/ß thereby inhibiting NF-κB activity in vivo. Cellular and nuclear levels of Ser(536)-phosphorylated (p65(536)) and Lys(310)-acetylated p65 subunit accompanied functional NF-κB activation during TMCH. RSK-1 phosphorylation at Thr(359)/Ser(363) in cellular/nuclear extracts and co-immunoprecipitation with cellular p65-NF-κB overlapped with p65(536) kinetics. Dietary pectin (6%) blocked NF-κB activity by blocking increases in p65 abundance and nuclear translocation thereby down-regulating CXCL-1/KC expression in the crypts. Thus, NF-κB activation persisted despite the lack of bacterial attachment to colonic mucosa beyond peak hyperplasia. The MEK/ERK/p38 pathway therefore seems to modulate sustained activation of NF-κB in colonic crypts in response to C. rodentium infection.
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Citrobacter rodentium , Colon/metabolismo , Enfermedades del Colon/metabolismo , Infecciones por Enterobacteriaceae/metabolismo , MAP Quinasa Quinasa 1/metabolismo , MAP Quinasa Quinasa 2/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Factor de Transcripción ReIA/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Animales , Quimiocina CXCL1/biosíntesis , Quimiocina CXCL1/genética , Colon/microbiología , Colon/patología , Enfermedades del Colon/genética , Enfermedades del Colon/microbiología , Enfermedades del Colon/patología , Infecciones por Enterobacteriaceae/genética , Flavonoides/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Hiperplasia/genética , Hiperplasia/metabolismo , Hiperplasia/microbiología , Quinasa I-kappa B/genética , Quinasa I-kappa B/metabolismo , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , MAP Quinasa Quinasa 1/antagonistas & inhibidores , MAP Quinasa Quinasa 1/genética , MAP Quinasa Quinasa 2/antagonistas & inhibidores , MAP Quinasa Quinasa 2/genética , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/genética , Ratones , Proteína Quinasa 3 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 3 Activada por Mitógenos/genética , Inhibidor NF-kappaB alfa , Fosforilación/efectos de los fármacos , Fosforilación/genética , Proteínas Quinasas S6 Ribosómicas 90-kDa/genética , Proteínas Quinasas S6 Ribosómicas 90-kDa/metabolismo , Factores de Tiempo , Factor de Transcripción ReIA/genética , Proteínas Quinasas p38 Activadas por Mitógenos/genéticaRESUMEN
Others and we previously showed that the vitamin D receptor (VDR) is subject to degradation by the 26S proteasome and that treatment with 1,25-dihydroxyvitamin D(3) (1,25D(3)) inhibited this degradation. In the present study, we found that in osteoblasts, but not in intestinal epithelial cells, the VDR was susceptible to degradation by the 26S proteasome. The subcellular site for degradation of the VDR in osteoblasts is the cytoplasm and the site for ligand-dependent protection of the VDR from the 26S proteasome is the chromatin. These direct relationships between nuclear localization and protection of the VDR from 26S proteasome degradation led us to hypothesize that the unoccupied cytoplasmic VDR is a substrate for polyubiquitination, which targets VDR for degradation by the 26S proteasome, and that nuclear localization has the ability to protect the VDR from polyubiquitination and degradation. To test these hypotheses, we used Cos-1 cells transfected with human VDR and histidine-tagged ubiquitin expression vectors. We found that unoccupied VDR was polyubiquitinated and that 1,25D(3) inhibited this modification. Mutations in the nuclear localization signal of VDR (R49W/R50G and K53Q/R54G/K55E) or in the dimerization interface of VDR with retinoid X receptor (M383G/Q385A) abolished the ability of 1,25D(3) to protect the VDR from polyubiquitination, although these mutations had no effect on the ligand-binding activity of VDR. Therefore, we concluded that in some cellular environments unoccupied cytoplasmic VDR is susceptible to polyubiquitination and proteasome degradation and that ligand-dependent heterodimerization and nuclear localization protect the VDR from these modifications.
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Núcleo Celular/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Receptores de Calcitriol/metabolismo , Animales , Línea Celular , Humanos , Hidrólisis , Transporte de Proteínas , UbiquitinaciónRESUMEN
Dietary calcium is believed to reduce colon cancer risk, but the mechanism by which this occurs is poorly understood. Employing the Citrobacter rodentium-induced transmissible murine colonic hyperplasia (TMCH) model, we previously showed that a high-calcium diet (hCa) significantly abrogated hyperplasia in the distal colons of NIH-Swiss mice. Here, we explored the mechanism of dietary protection by hCa by analyzing the expression of genes involved in the regulation of Ca uptake/flux in the intestinal epithelium, including the Ca-sensing receptor, vitamin D receptor, Ca binding protein, and transient receptor potential cation channels, subfamily V, members 5 and 6 (TRPV5/6). Interestingly, while TRPV6 expression increased significantly during TMCH, the expression of the other gene products was unchanged. This elevated TRPV6 expression was significantly abrogated by a hCa diet. Immunofluorescence revealed apical membrane localization of TRPV6 in the normal colon, whereas during TMCH we observed intense apical pole and cytoplasmic staining along the entire longitudinal crypt axis, including the expanded proliferating zone. The hCa diet reversed this effect. In humans, overexpression of TRPV6 was associated with early-stage colon cancer, and in colon carcinoma cells, inhibition of TRPV6 expression by small interfering RNA inhibited their proliferation and induced apoptosis. TRPV6 small interfering RNA also diminished the transcriptional activity of the calcium-dependent nuclear factors in activated T cells. Thus the aberrant overexpression of TRPV6 contributes to colonic crypt hyperplasia in mice and to colon cancer cell proliferation in humans. Therefore, it is likely that suppression of TRPV6 by a hCa diet is required for its protective effects in the colon.
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Canales de Calcio/metabolismo , Calcio de la Dieta/farmacología , Enfermedades del Colon/prevención & control , Regulación de la Expresión Génica/efectos de los fármacos , Canales Catiónicos TRPV/metabolismo , Animales , Calcitriol , Canales de Calcio/genética , Proliferación Celular , Citrobacter rodentium , Colon/citología , Colon/metabolismo , Colon/patología , Enfermedades del Colon/metabolismo , Dieta , Infecciones por Enterobacteriaceae/tratamiento farmacológico , Ratones , Canales Catiónicos TRPV/genéticaRESUMEN
Eight new side-chain allylic, benzylic, and propargylic ether analogs of the natural hormone calcitriol have been rationally designed and easily synthesized. Three of these 23-oxa ether analogs lacking the typical side-chain OH group are more antiproliferative in vitro and desirably less calcemic in vivo than the natural hormone. One of these three 23-oxa analogs has transcriptional potency almost as high as that of calcitriol, even though it binds to the human vitamin D receptor only about 1% as well as calcitriol.
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Calcitriol/análogos & derivados , Calcitriol/síntesis química , Calcio/orina , Proliferación Celular/efectos de los fármacos , Alquinos/síntesis química , Alquinos/farmacología , Compuestos Alílicos/síntesis química , Compuestos Alílicos/farmacología , Animales , Compuestos de Bencilo/síntesis química , Compuestos de Bencilo/farmacología , Unión Competitiva , Calcitriol/farmacología , Canales de Calcio/biosíntesis , Canales de Calcio/genética , Línea Celular , Duodeno/metabolismo , Femenino , Humanos , Queratinocitos/citología , Queratinocitos/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Ratas , Receptores de Calcitriol/metabolismo , Estereoisomerismo , Esteroide Hidroxilasas/biosíntesis , Esteroide Hidroxilasas/genética , Relación Estructura-Actividad , Canales Catiónicos TRPV/biosíntesis , Canales Catiónicos TRPV/genética , Transcripción Genética/efectos de los fármacos , Vitamina D3 24-HidroxilasaRESUMEN
Bone is the primary site of metastases in advanced androgen-independent prostate cancer. These metastases are primarily bone-forming, although the presence of osteolytic response has also been reported. Bone-homing therapy is a strategy based on the popular seed-and-soil relationship between the epithelial malignant cells and the bone stroma. Calcitriol (1,25-dihydroxyvitamin D3) and its synthetic analogs (deltanoids) are drugs that have a direct effect on both the skeleton and the invading metastatic cells and, therefore, are considered useful in the treatment of advanced prostate cancer. In this article, I review the nature of the response induced by the malignant cells in the bone (bone formation or bone resorption) and how it affects the outcome of a vitamin D analog treatment in preclinical models of metastatic bone disease.
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Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/secundario , Vitamina D/análogos & derivados , Animales , Neoplasias Óseas/metabolismo , Neoplasias Óseas/patología , Calcitriol/análogos & derivados , Calcitriol/uso terapéutico , Línea Celular Tumoral , Humanos , Masculino , Ratones , Modelos Biológicos , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Osteoblastos/patología , Osteólisis/tratamiento farmacológico , Osteólisis/metabolismo , Osteólisis/patología , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/metabolismo , Receptores de Calcitriol/metabolismo , Vitamina D/uso terapéuticoRESUMEN
1,25(OH)(2)D(3) regulates calcium homeostasis through its actions in the intestine, bone, and kidney. These actions are mediated through the VDR. To determine if VDR's actions in the proximal small intestine can sufficiently restore calcium homeostasis, we generated transgenic mice expressing hVDR exclusively in the duodenum of mVDR-null mice by using the adenosine deaminase enhancer (hVDR+/mVDR-null). Unlike wild-type mice, hVDR+/mVDR-null mice express hVDR and VDR target genes only in the proximal small intestine. Despite having functional hVDR in the proximal small intestine, hVDR+/mVDR-null mice were hypocalcaemic when fed a normal rodent diet at weaning, like mVDR-null mice fed the same diet. The hypocalcemia in these mice is prevented if they are given the rescue diet before weaning. However, when 90-day-old rachitic mice were fed a rescue diet, serum calcium improved in hVDR+/mVDR-null mice, but not in mVDR-null mice. In conclusion, transgenic hVDR in the proximal small intestine of hVDR+/mVDR-null mice was transcriptionally active and regulated calcium absorption, but VDR actions elsewhere in the intestine are probably necessary to support adequate calcium homeostasis. In addition, hVDR+/mVDR-null mice responded better to the late rescue diet suggesting that expression of VDR in the proximal small intestine protected the calcium absorbing machinery from age-dependent decline.
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Envejecimiento/fisiología , Calcio/sangre , Duodeno/metabolismo , Regulación de la Expresión Génica , Receptores de Calcitriol/deficiencia , Receptores de Calcitriol/metabolismo , Animales , Humanos , Ratones , Ratones Noqueados , Receptores de Calcitriol/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Esteroide Hidroxilasas/genética , Vitamina D3 24-HidroxilasaRESUMEN
We report new analogs of 1alpha,25-dihydroxyvitamin D(3) (1) in three categories. First, design and synthesis of ligands for a mutant vitamin D receptor (VDR)(Arg274Leu), which possess proper functional groups at both C1alpha and C2alpha positions of 1 to study the biological activity of the mutant VDR. Among our synthetic analogs, 1alpha-methyl-2alpha-(3-hydroxypropyl)-25-hydroxyvitamin D(3) (8) showed 7.3-fold greater transcriptional activity for the VDR(Arg274Leu) than that of 1. Next, we examined the antiproliferative activity of 2-substituted 19-norvitamin D(3) analogs on an immortalized normal prostate cell line, PZ-HPV-7, and we found MART 10 (14) showed the activity even at very low concentration of 10(-10) to 10(-11)M. We also synthesized 25-hydroxy-19-norvitamin D(3) (13) using Julia-type olefination to connect between the C5 and C6 positions, effectively, to test it as a prohormone type agent for antiprostate diseases. Synthesized compound 13 showed potent antiproliferative activity in PZ-HPV-7, which has high 1alpha-hydroxylase activity. Finally, we describe design and synthesis of a new TEI-9647 analog, 2alpha-(3-hydroxypropoxy)-24-propyl-25-dehydro-1alpha-hydroxyvitamin D(3)-26,23-lactone (17), which showed the strongest VDR antagonism. Its IC(50) value is 7.4pM to inhibit differentiation of HL-60 cells induced by 10nM of 1.
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Enfermedad , Salud , Vitamina D/análogos & derivados , Vitamina D/síntesis química , Aminoácidos/química , Animales , Línea Celular , Proliferación Celular/efectos de los fármacos , Humanos , Enlace de Hidrógeno , Ligandos , Masculino , Estructura Molecular , Osteítis Deformante/metabolismo , Osteítis Deformante/patología , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Vitamina D/química , Vitamina D/farmacologíaRESUMEN
Three new Vitamin D analogs 3-5 incorporating a -CHF(2) group as an -OH surrogate have been prepared. Two of these new analogs (3 and 5) are strongly antiproliferative toward murine keratinocytes and are approximately 50 times less calciuric in vivo than the natural hormone calcitriol. The transcriptional activity of the 25-CHF(2) analog 3 is higher than that of the 1-CHF(2) analog 4.
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Productos Biológicos/química , Productos Biológicos/farmacología , Diseño de Fármacos , Flúor/química , Hormonas/química , Vitamina D/análogos & derivados , Animales , Productos Biológicos/síntesis química , Calcio/orina , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Hormonas/síntesis química , Hormonas/farmacología , Metilación , Ratones , Estructura Molecular , Ratas , Vitamina D/síntesis química , Vitamina D/química , Vitamina D/farmacologíaRESUMEN
We examined the effects of 1,25 dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) on the distribution and mobility of the vitamin D receptor (VDR) in the enterocyte-like Caco-2 cell. Confocal microscopy showed that a green fluorescent protein-vitamin D receptor (GFP-VDR) fusion protein is predominantly nuclear (58%) and it does not associate with the apical or basolateral membrane of proliferating or polarized, differentiated cells. In contrast to the previously studied cell types, neither endogenous VDR nor GFP-VDR levels accumulate in the nucleus following 1,25(OH)(2)D(3) treatment (100 nM, 30 min). However, in nuclear photobleaching experiments nuclear GFP-VDR import was significantly increased by 1,25(OH)(2)D(3) during both an early (0-5 min) and later (30-35 min) period (20% per 5 min). Compared to the natural ligand, nuclear import of GFP-VDR was 60% lower in cells treated with the 1,25(OH)(2)D(3) analog, 1-alpha-fluoro-16-ene-20-epi-23-ene-26,27-bishomo-25-hydroxyvitamin D(3) (Ro-26-9228, 5 min, 100 nM). Downstream events like ligand-induced association of VDR with chromatin at 1 h and the accumulation of CYP24 mRNA were significantly lower in Ro-26-9228 treated cells compared to 1,25(OH)(2)D(3) (60 and 95% lower, respectively). Collectively our data are consistent with a role for ligand-induced nuclear VDR import in receptor activation. In addition, ligand-dependent VDR nuclear import appears to be balanced by export, thus accounting for the lack of nuclear VDR accumulation even when VDR import is significantly elevated.
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Colon/metabolismo , Receptores de Calcitriol/metabolismo , Secuencia de Bases , Células CACO-2 , Calcitriol/farmacología , Colon/citología , Colon/efectos de los fármacos , Cartilla de ADN , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Transporte de Proteínas , Transcripción GenéticaRESUMEN
Replacing the 1alpha-OH group of the natural hormone 1alpha,25-dihydroxyvitamin D(3) (calcitriol) by a 1alpha-CHF(2) group and incorporating a potentiating side chain produced two new hybrid analogs 6 and 7. Both of these two hybrid analogs are as transcriptionally active as calcitriol and are strongly antiproliferative in vitro but are low-calcemic in vivo.
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Antineoplásicos/síntesis química , Calcitriol/análogos & derivados , Calcitriol/síntesis química , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Calcitriol/química , Calcitriol/farmacología , Calcio/orina , Línea Celular , Proliferación Celular/efectos de los fármacos , Femenino , Humanos , Queratinocitos/citología , Queratinocitos/efectos de los fármacos , Ratones , Ensayo de Unión Radioligante , Ratas , Ratas Sprague-Dawley , Receptores de Calcitriol/metabolismo , Estereoisomerismo , Relación Estructura-Actividad , Transcripción GenéticaRESUMEN
This study aimed to investigate whether any physiological changes might have a clinically significant effect on function in sedentary, institutionalized, older adults treated by a passive training program. A total of 18 subjects (mean age 60.7 +/- 3.4) with intellectual disability (ID) participated. We measured SpO2 (arterial oxygen saturation) before, during, and after passive training, and used Barthel Index to measure daily living activities. The general trend indicated that inactive people with ID evidenced a continual increase in SpO2% levels and some functional gains during passive treatment, with superiority to manual passive treatment compared to mechanical active passive training. For current clinical practice, most sedentary patients who experience clinically significant deconditioning and desaturation can benefit from passive treatment.
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Pierna/fisiología , Terapia Pasiva Continua de Movimiento , Oxígeno/sangre , Actividades Cotidianas , Anciano , Presión Sanguínea , Femenino , Frecuencia Cardíaca , Humanos , Institucionalización , Discapacidad Intelectual , Masculino , Persona de Mediana Edad , Actividad Motora , Rango del Movimiento ArticularRESUMEN
Prostate cancer metastasizes almost exclusively into the bone whereby it induces primarily an osteoblastic response. Non-calcemic vitamin D analogs have been shown to inhibit proliferation of prostate cancer cells in culture and inhibit their growth as subcutaneous xenografts in mice. However, their effect on prostate cancer cell growth in the bone has not been examined. In the present study, we inoculated the osteoblastic prostate cancer cell line MDA-PCa 2b into the bone of male SCID mice and examined the effect of the low-calcemic hybrid analog 1alpha-hydroxymethyl-16-ene-26,27-bishomo-25-hydroxy vitamin D(3) (JK-1626-2) on their ability to induce bone lesions. We found that 7 weeks after inoculation of MDA-PCa 2b cells, 90% of the mice in the vehicle-treated group had significant bone lesions that were detectable by micro-computed tomography and characterized by thickening of the cortical bone and ossification of the epiphysis. Only 30% of the mice in the analog-treated group (daily injections of 4microg/kg, 5 days/week for up to 7 weeks) had detectable bone lesions. Histological examination of the decalcified tumor-bearing bones has shown that tumor cells completely replaced the bone marrow in the diaphysis, and destroyed the trabecular bone in the metaphysis in 90% of the vehicle-treated mice. In contrast, the metaphysis of 60% of analog-treated mice appeared normal, although tumor cells were still found in the diaphysis of 70% of the bones in the analog-treated group. There was no evidence of hypercalcemia in any of the analog-treated mice. In a co-culture, MDA-PCa 2b cells induced a profound mitogenic response in osteoblasts followed by enhanced differentiation. However, in the presence of the analog the mitogenic response of the osteoblasts to the malignant cells was significantly attenuated. These experiments led to the hypothesis that, in vivo, JK-1626-2 prevented the metastatic bone lesions by inhibiting the mitogenic response of osteoblasts to growth factors produced by MDA-PCa 2b cells.
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Calcifediol/análogos & derivados , Calcifediol/farmacología , Osteoblastos/efectos de los fármacos , Osteoblastos/patología , Neoplasias de la Próstata/patología , Neoplasias de la Próstata/prevención & control , Animales , Calcio/metabolismo , Células Cultivadas , Progresión de la Enfermedad , Masculino , Ratones , Estructura MolecularRESUMEN
1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] has been reported to stimulate lung maturity, alveolar type II cell differentiation, and pulmonary surfactant synthesis in rat lung. We hypothesized that 1,25(OH)(2)D(3) stimulates expression of surfactant protein-A (SP-A), SP-B, and SP-C in human fetal lung and type II cells. We found that immunoreactive vitamin D receptor was detectable in fetal lung tissue and type II cells only when incubated with 1,25(OH)(2)D(3). 1,25(OH)(2)D(3) significantly decreased SP-A mRNA in human fetal lung tissue but did not significantly decrease SP-A protein in the tissue. In type II cells, 1,25(OH)(2)D(3) alone had no significant effect on SP-A mRNA or protein levels but reduced SP-A mRNA and protein in a dose-dependent manner when the cells were incubated with cAMP. SP-A mRNA levels in NCI-H441 cells, a nonciliated bronchiolar epithelial (Clara) cell line, were decreased in a dose-dependent manner in the absence or presence of cAMP. 1,25(OH)(2)D(3) had no significant effect on SP-B mRNA levels in lung tissue but increased SP-B mRNA and protein levels in type II cells incubated in the absence or presence of cAMP. Expression of SP-C mRNA was unaffected by 1,25(OH)(2)D(3) in lung tissue incubated +/- cAMP. These results suggest that regulation of surfactant protein gene expression in human lung and type II cells by 1,25(OH)(2)D(3) is not coordinated; 1,25(OH)(2)D(3) decreases SP-A mRNA and protein levels in both fetal lung tissue and type II cells, increases SP-B mRNA and protein levels only in type II cells, and has no effect on SP-C mRNA levels.
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Alveolos Pulmonares/fisiología , Proteínas Asociadas a Surfactante Pulmonar/genética , Vitamina D/análogos & derivados , Adenocarcinoma , Secuencia de Bases , Sitios de Unión/genética , Línea Celular Tumoral , Expresión Génica/efectos de los fármacos , Humanos , Neoplasias Pulmonares , Datos de Secuencia Molecular , Técnicas de Cultivo de Órganos , Alveolos Pulmonares/citología , Alveolos Pulmonares/embriología , Proteína A Asociada a Surfactante Pulmonar/genética , Proteína B Asociada a Surfactante Pulmonar/genética , Proteína C Asociada a Surfactante Pulmonar/genética , ARN Mensajero/análisis , Vitamina D/farmacologíaRESUMEN
A series 5-8 of 1- and 3-CH(2)OH 19-nor analogs of the hormone calcitriol (1) has been prepared. Surprisingly, 19-nor 1alpha-CH(2)OH analog 5a is more antiproliferative at 100 nM concentration than the corresponding regioisomeric analog 6a with the natural 1alpha-OH group, and 1alpha-CH(2)OH hybrid analog 7a is similar in antiproliferative potency to calcitriol (1) even at low nanomolar concentrations.
Asunto(s)
Antineoplásicos/síntesis química , Calcitriol/análogos & derivados , Animales , Antineoplásicos/farmacología , Calcitriol/síntesis química , Calcitriol/farmacología , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Queratinocitos/efectos de los fármacos , Ratones , Relación Estructura-ActividadRESUMEN
Prepared from a commercial prostaglandin building block, novel vitamin D3 analogs with a contracted five-membered A-ring were designed and synthesized to mimic the A-ring diol structure of the natural hormone 1alpha,25-dihydroxyvitamin D3. Prepared from commercial 1,4-cyclohexanedione, a structurally simplified analog was designed and synthesized in which a suitably oriented primary allylic hydroxyl group at the C-2 position might be a surrogate for the biologically important 1alpha-OH in the natural hormone.
Asunto(s)
Vitamina D/análogos & derivados , Animales , Células COS , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Chlorocebus aethiops , Humanos , Queratinocitos/efectos de los fármacos , Relación Estructura-Actividad , Vitamina D/síntesis química , Vitamina D/química , Vitamina D/farmacologíaRESUMEN
The 25-hydroxyvitamin D(3) (25-OH-D(3)) is a nontoxic and low-affinity vitamin D receptor (VDR)-binding metabolic precursor of 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)]. We hypothesized that covalent attachment of a 25-OH-D(3) analog to the hormone-binding pocket of VDR might convert the latter into transcriptionally active holo-form, making 25-OH-D(3) biologically active. Furthermore, it might be possible to translate the nontoxic nature of 25-OH-D(3) into its analog. We showed earlier that 25-hydroxyvitamin D(3)-3-bromoacetate (25-OH-D(3)-3-BE) alkylated the hormone-binding pocket of VDR. In this communication we describe that 10(-6) mol/L of 25-OH-D(3)-3-BE inhibited the growth of keratinocytes, LNCaP, and LAPC-4 androgen-sensitive and PC-3 and DU145 androgen-refractory prostate cancer cells, and PZ-HPV-7 immortalized normal prostate cells with similar or stronger efficacy as 1,25(OH)(2)D(3). But its effect was strongest in LNCaP, PC-3, LAPC-4, and DU145 cells. Furthermore, 25-OH-D(3)-3-BE was toxic to these prostate cancer cells and caused these cells to undergo apoptosis as shown by DNA-fragmentation and caspase-activation assays. In a reporter assay with COS-7 cells, transfected with a 1alpha,25-dihydroxyvitamin D(3)-24-hydroxylase (24-OHase)-construct and VDR-expression vector, 25-OH-D(3)-3-BE induced 24-OHase promoter activity. In a "pull down assay" with PC-3 cells, 25-OH-D(3)-3-BE induced strong interaction between VDR and general transcription factors, retinoid X receptor, and GRIP-1. Collectively, these results strongly suggested that the cellular effects of 25-OH-D(3)-3-BE were manifested via 1,25(OH)(2)D(3)/VDR signaling pathway. A toxicity study in CD-1 mice showed that 166 microg/kg of 25-OH-D(3)-3-BE did not raise serum-calcium beyond vehicle control. Collectively, these results strongly suggested that 25-OH-D(3)-3-BE has a strong potential as a therapeutic agent for androgen-sensitive and androgen-refractory prostate cancer.
Asunto(s)
Apoptosis/efectos de los fármacos , Calcitriol/análogos & derivados , Calcitriol/farmacología , Proliferación Celular/efectos de los fármacos , Neoplasias Hormono-Dependientes/tratamiento farmacológico , Neoplasias de la Próstata/tratamiento farmacológico , 25-Hidroxivitamina D3 1-alfa-Hidroxilasa/genética , Animales , Células COS , Proteínas Portadoras/metabolismo , Caspasas/metabolismo , Cloranfenicol O-Acetiltransferasa , Chlorocebus aethiops , Relación Dosis-Respuesta a Droga , Activación Enzimática/efectos de los fármacos , Humanos , Queratinocitos/citología , Queratinocitos/efectos de los fármacos , Masculino , Ratones , Neoplasias Hormono-Dependientes/patología , Proteínas del Tejido Nervioso/metabolismo , Regiones Promotoras Genéticas , Próstata/citología , Próstata/efectos de los fármacos , Neoplasias de la Próstata/patología , Receptores de Calcitriol/metabolismo , Receptores X Retinoide/metabolismo , Timidina/metabolismo , Células Tumorales CultivadasRESUMEN
A dozen 24-sulfoximine analogues of the hormone 1alpha,25-dihydroxyvitamin D(3) were prepared, differing not only at the stereogenic sulfoximine stereocenter but also at the A-ring. Although these sulfoximines were not active transcriptionally and were only very weakly antiproliferative, some of them are powerful hydroxylase enzyme inhibitors. Specifically, 24-(S)-NH phenyl sulfoximine 3a is an extremely potent CYP24 inhibitor (IC(50) = 7.4 nM) having low calcemic activity. In addition, this compound shows high selectivity toward the CYP24 enzyme in comparison to CYP27A1 (IC(50) > 1000 nM) and CYP27B (IC(50) = 554 nM).