Pulling rank with RNA: RANKL promotes the association of PGC-1ß/RNA complexes with NCoR/HDAC3 to activate gene expression in osteoclasts.

In this issue, Abe et al1 report a novel mechanism by which RANKL stimulates osteoclast differentiation and bone resorption through non-coding RNAs that bind PGC-1ß and convert the NCoR/HDAC3 co-repressor complex into a co-activator of AP-1- and NFκB-regulated genes.

Comprehensive analysis of the proximity-dependent nuclear interactome for the oncoprotein NOTCH1 in live cells.

Notch signaling plays a critical role in cell fate decisions in all cell types. Furthermore, gain-of-function mutations in NOTCH1 have been uncovered in many human cancers. Disruption of Notch signaling has recently emerged as an attractive disease treatment strategy. However, the nuclear interaction landscape of the oncoprotein NOTCH1 remains largely unexplored. We therefore employed here a proximity-dependent biotin identification approach to identify in vivo protein associations with the nuclear Notch1 intracellular domain in live cells. We identified a large set of previously reported and unreported proteins that associate with NOTCH1, including general transcription and elongation factors, DNA repair and replication factors, coactivators, corepressors, and components of the NuRD and SWI/SNF chromatin remodeling complexes. We also found that Notch1 intracellular domain associates with protein modifiers and components of other signaling pathways that may influence Notch signal transduction and protein stability such as USP7. We further validated the interaction of NOTCH1 with histone deacetylase 1 or GATAD2B using protein network analysis, proximity-based ligation, in vivo cross-linking and coimmunoprecipitation assays in several Notch-addicted cancer cell lines. Through data mining, we also revealed potential drug targets for the inhibition of Notch signaling. Collectively, these results provide a valuable resource to uncover the mechanisms that fine-tune Notch signaling in tumorigenesis and inform therapeutic targets for Notch-addicted tumors.

Inflammatory Processes Affecting Bone Health and Repair.

PURPOSE OF REVIEW: The purpose of this article is to review the current understanding of inflammatory processes on bone, including direct impacts of inflammatory factors on bone cells, the effect of senescence on inflamed bone, and the critical role of inflammation in bone pain and healing. RECENT FINDINGS: Advances in osteoimmunology have provided new perspectives on inflammatory bone loss in recent years. Characterization of so-called inflammatory osteoclasts has revealed insights into physiological and pathological bone loss. The identification of inflammation-associated senescent markers in bone cells indicates that therapies that reduce senescent cell burden may reverse bone loss caused by inflammatory processes. Finally, novel studies have refined the role of inflammation in bone healing, including cross talk between nerves and bone cells. Except for the initial stages of fracture healing, inflammation has predominately negative effects on bone and increases fracture risk. Eliminating senescent cells, priming the osteo-immune axis in bone cells, and alleviating pro-inflammatory cytokine burden may ameliorate the negative effects of inflammation on bone.

Statin-mediated cholesterol depletion exerts coordinated effects on the alterations in rat vascular smooth muscle cell biomechanics and migration.

KEY POINTS: This study demonstrates and evaluates the changes in rat vascular smooth muscle cell biomechanics following statin-mediated cholesterol depletion. Evidence is presented to show correlated changes in migration and adhesion of vascular smooth muscle cells to extracellular matrix proteins fibronectin and collagen. Concurrently, integrin α5 expression was enhanced but not integrin α2. Atomic force microscopy analysis provides compelling evidence of coordinated reduction in vascular smooth muscle cell stiffness and actin cytoskeletal orientation in response to statin-mediated cholesterol depletion. Proof is provided that statin-mediated cholesterol depletion remodels total vascular smooth muscle cell cytoskeletal orientation that may additionally participate in altering ex vivo aortic vessel function. It is concluded that statin-mediated cholesterol depletion may coordinate vascular smooth muscle cell migration and adhesion to different extracellular matrix proteins and regulate cellular stiffness and cytoskeletal orientation, thus impacting the biomechanics of the cell. ABSTRACT: Not only does cholesterol induce an inflammatory response and deposits in foam cells at the atherosclerotic plaque, it also regulates cellular mechanics, proliferation and migration in atherosclerosis progression. Statins are HMG-CoA reductase inhibitors that are known to inhibit cellular cholesterol biosynthesis and are clinically prescribed to patients with hypercholesterolemia or related cardiovascular conditions. Nonetheless, the effect of statin-mediated cholesterol management on cellular biomechanics is not fully understood. In this study, we aimed to assess the effect of fluvastatin-mediated cholesterol management on primary rat vascular smooth muscle cell (VSMC) biomechanics. Real-time measurement of cell adhesion, stiffness, and imaging were performed using atomic force microscopy (AFM). Cellular migration on extra cellular matrix (ECM) protein surfaces was studied by time-lapse imaging. The effect of changes in VSMC biomechanics on aortic function was assessed using an ex vivo myograph system. Fluvastatin-mediated cholesterol depletion (-27.8%) lowered VSMC migration distance on a fibronectin (FN)-coated surface (-14.8%) but not on a type 1 collagen (COL1)-coated surface. VSMC adhesion force to FN (+33%) and integrin α5 expression were enhanced but COL1 adhesion and integrin α2 expression were unchanged upon cholesterol depletion. In addition, VSMC stiffness (-46.6%) and ex vivo aortic ring contraction force (-40.1%) were lowered and VSMC actin cytoskeletal orientation was reduced (-24.5%) following statin-mediated cholesterol depletion. Altogether, it is concluded that statin-mediated cholesterol depletion may coordinate VSMC migration and adhesion to different ECM proteins and regulate cellular stiffness and cytoskeletal orientation, thus impacting the biomechanics of the cell and aortic function.

Hdac1 and Hdac2 positively regulate Notch1 gain-of-function pathogenic signaling in committed osteoblasts of male mice.

BACKGROUND: Skeletal development requires precise extrinsic and intrinsic signals to regulate processes that form and maintain bone and cartilage. Notch1 is a highly conserved signaling receptor that regulates cell fate decisions by controlling the duration of transcriptional bursts. Epigenetic molecular events reversibly modify DNA and histone tails by influencing the spatial organization of chromatin and can fine-tune the outcome of a Notch1 transcriptional response. Histone deacetylase 1 and 2 (HDAC1 and HDAC2) are chromatin modifying enzymes that mediate osteoblast differentiation. While an HDAC1-Notch interaction has been studied in vitro and in Drosophila, its role in mammalian skeletal development and disorders is unclear. Osteosclerosis is a bone disorder with an abnormal increase in the number of osteoblasts and excessive bone formation. METHODS: Here, we tested whether Hdac1/2 contribute to the pathogenesis of osteosclerosis in a murine model of the disease owing to conditionally cre-activated expression of the Notch1 intracellular domain in immature osteoblasts. RESULTS: Importantly, selective homozygous deletions of Hdac1/2 in osteoblasts partially alleviate osteosclerotic phenotypes (Col2.3kb-Cre; TGRosaN1ICD/+ ; Hdac1flox/flox ; Hdac2flox/flox ) with a 40% decrease in bone volume and a 22% decrease in trabecular thickness in 4 weeks old when compared to male mice with heterozygous deletions of Hdac1/2 (Col2.3 kb-Cre; TGRosaN1ICD/+ ; Hdac1flox/+ ; Hdac2flox/+ ). Osteoblast-specific deletion of Hdac1/2 in male and female mice results in no overt bone phenotype in the absence of the Notch1 gain-of-function (GOF) allele. CONCLUSIONS: These results provide evidence that Hdac1/2 contribute to Notch1 pathogenic signaling in the mammalian skeleton. Our study on epigenetic regulation of Notch1 GOF-induced osteosclerosis may facilitate further mechanistic studies of skeletal birth defects caused by Notch-related GOF mutations in human patients, such as Adams-Oliver disease, congenital heart disease, and lateral meningocele syndrome.

Precise detection of a murine germline mutation of the Notch3 gene associated with kyphosis and developmental disorders.

OBJECTIVE: Humpback (hpbk) mice harbor a pathogenic mutation in the Notch3 gene and can serve as a beneficial animal model for investigating human myopathy, kyphosis, and developmental disorders, including lateral meningocele syndrome. Detection of the point mutation in hpbk mice is important for maintaining strains and scrutinizing genetic rescues, especially considering that homozygous mice are infertile and indistinguishable from their littermates at a young age. This study aimed for the development of a novel, precise, and time-saving genotyping method to identify the mutation in hpbk mice. MATERIALS AND METHODS: In order to study the hpbk mouse line, we describe how we applied several tools, including quantitative polymerase chain reaction (qPCR), multiplex tetra-primer amplification-refractory mutation system (ARMS-PCR) and Sanger sequencing, toward the recognition of heterozygous and homozygous mice. RESULTS: The Notch3 mutation was clearly identified using qPCR and ARMS assays, but the latter was a more precise and cost-effective approach. The lengths of the ARMS-PCR amplicons are 210 bp and 164 bp for the wild-type and hpbk alleles, respectively. Moreover, the genotyping results for each mouse were corroborated by Sanger DNA sequencing. CONCLUSION: Our newly developed PCR-based ARMS system affords a swift and precise way to genotype the hpbk mice. ARMS-PCR does not rely on any advanced equipment and is useful as a genotyping method for other model organisms that harbor a pathogenic variant.

Selective Targeting of Class I Histone Deacetylases in a Model of Human Osteosarcoma.

Dysregulation of histone deacetylases (HDACs) is associated with the pathogenesis of human osteosarcoma, which may present an epigenetic vulnerability as well as a therapeutic target. Domatinostat (4SC-202) is a next-generation class I HDAC inhibitor that is currently being used in clinical research for certain cancers, but its impact on human osteosarcoma has yet to be explored. In this study, we report that 4SC-202 inhibits osteosarcoma cell growth in vitro and in vivo. By analyzing cell function in vitro, we show that the anti-tumor effect of 4SC-202 involves the combined induction of cell-cycle arrest at the G2/M phase and apoptotic program, as well as a reduction in cell invasion and migration capabilities. We also found that 4SC-202 has little capacity to promote osteogenic differentiation. Remarkably, 4SC-202 revised the global transcriptome and induced distinct signatures of gene expression in vitro. Moreover, 4SC-202 decreased tumor growth of established human tumor xenografts in immunodeficient mice in vivo. We further reveal key targets regulated by 4SC-202 that contribute to tumor cell growth and survival, and canonical signaling pathways associated with progression and metastasis of osteosarcoma. Our study suggests that 4SC-202 may be exploited as a valuable drug to promote more effective treatment of patients with osteosarcoma and provide molecular insights into the mechanism of action of class I HDAC inhibitors.

Development and characterization of the novel human osteosarcoma cell line COS-33 with sustained activation of the mTOR pathway.

Outcomes have not improved for metastatic osteosarcoma for several decades. In part, this failure to develop better therapies stems from a lack of understanding of osteosarcoma biology, given the rarity of the disease and the high genetic heterogeneity at the time of diagnosis. We report here the successful establishment of a new human osteosarcoma cell line, COS-33, from a patient-derived xenograft and demonstrate retention of the biological features of the original tumor. We found high mTOR signaling activity in the cultured cells, which were sensitive to a small molecule inhibitor, rapamycin, a suppressor of the mTOR pathway. Suppressed mTOR signaling after treatment with rapamycin was confirmed by decreased phosphorylation of the S6 ribosomal protein. Increasing concentrations of rapamycin progressively inhibited cell proliferation in vitro. We observed significant inhibitory effects of the drug on cell migration, invasion, and colony formation in the cultured cells. Furthermore, we found that only a strong osteogenic inducer, bone morphogenetic protein-2, promoted the cells to differentiate into mature mineralizing osteoblasts, indicating that the COS-33 cell line may have impaired osteoblast differentiation. Grafted COS-33 cells exhibited features typical of osteosarcoma, such as production of osteoid and tumorigenicity in vivo. In addition, we revealed that the COS-33 cell line retained a complex karyotype, a homozygous deletion of the TP53 gene, and typical histological features from its original tumor. Our novel cellular model may provide a valuable platform for studying the etiology and molecular pathogenesis of osteosarcoma as well as for testing novel drugs for future genome-informed targeted therapy.

Targeting the Wnt/ß-catenin pathway in human osteosarcoma cells.

Aberrant activation of Wnt signaling has been implicated in human osteosarcoma, which may provide a genetic vulnerability that can be targeted in osteosarcoma treatment. To test whether Wnt activation is necessary for osteosarcoma growth, colony formation, invasion, and metastasis, we treated human osteosarcoma cells with a small molecule inhibitor of Wnt/ß-catenin, PRI-724, which suppresses Wnt/ß-catenin-mediated transcription. We found increased protein levels of endogenous active-ß-catenin in five human osteosarcoma cell lines. Treatment with PRI-724 was sufficient to inhibit human osteosarcoma 143B and SJSA-1 cell proliferation. Suppressed Wnt signaling was confirmed by decreased protein levels of the Wnt target Cyclin D1. Furthermore, we revealed significant inhibitory effects on cell migration, invasion, and colony formation in the human osteosarcoma cells. Using deposited data from next generation sequencing studies, we analyzed somatic mutations and gene expression of components in the Wnt/ß-catenin pathway. We found somatic mutations and upregulated gene expression of many components in the Wnt/ ß-catenin pathway, indicating activated Wnt signaling. Taken together, our results illustrate the critical role of Wnt/ß-catenin signaling in human osteosarcoma pathogenesis and growth, as well as the therapeutic potential of Wnt inhibitors in the treatment of human osteosarcoma.

5,6,7,3',4',5'-Hexamethoxyflavone inhibits growth of triple-negative breast cancer cells via suppression of MAPK and Akt signaling pathways and arresting cell cycle.

Natural components continue to be an important source for the discovery and development of novel anticancer agents. Polymethoxyflavones are a class of flavonoids found in citrus fruits and medicinal plants used in traditional medicine. In the present study, the anticancer activity of the well-known nobiletin (5,6,7,8,3',4'-hexamethoxyflavone) was compared against its less studied structural isomer 5,6,7,3',4',5'-hexamethoxyflavone. These compounds were evaluated on the Hs578T triple-negative breast cancer cell line and its more migratory subclone Hs578Ts(i)8. 5,6,7,3',4',5'-hexamethoxyflavone was found to be less toxic than nobiletin, while a similar growth inhibitory effect was observed after 72 h. Additionally, 5,6,7,3',4',5'-hexamethoxyflavone arrested the cell cycle in the G2/M phase, while no effect was observed on apoptosis or the migratory behavior of these cells. Furthermore, mechanistic studies revealed that the growth inhibition was concomitant with reduced phosphorylation levels of signaling molecules in the MAPK and Akt pathways as well as cell cycle regulators, involved in regulating cell proliferation, survival and cell cycle. In summary, the present study is the first to report on the anticancer activities of 5,6,7,3',4',5'-hexamethoxyflavone and to provide evidence that this flavone could have a greater potential than nobiletin for prevention or treatment of triple- negative breast cancer.

FAK tyrosine 407 organized with integrin αVß5 in Hs578Ts(i)8 advanced triple-negative breast cancer cells.

Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase known to promote cell migration and invasiveness. Overexpression and increased activity of FAK are closely associated with metastatic breast tumors and are linked to poor prognosis. This study discovered an inverse correlation between FAK activity and migratory and invasive behavior. We show decreased phosphorylation levels of FAK at tyrosine residues 397 and 861, and most prominently at Y407, in the more invasive Hs578Ts(i)8 subclone of the Hs578T breast cancer progression model. There is limited information available on FAK Y407, and here we demonstrate its presence in triple-negative breast cancer (TNBC) cell lines. Furthermore, our studies propose that localization of FAK Y407, rather than FAK expression and overall FAK Y407 phosphorylation levels, is crucial for the control of cell motility. FAK Y407 is found extensively at the cell periphery in focal adhesion-like structures at each end of actin stress fibers and organized with integrin αVß5 receptors, linking the αVß5 integrin-mediated migratory behavior of Hs578Ts(i)8 cells to FAK Y407. These data suggest that subcellular localization, next to expression and activity levels, are important for understanding TNBC progression. Such an approach opens new avenues for further studies and may provide novel insight for the classification of TNBC and facilitate the discovery of effective biomarkers for diagnosis and therapy of TNBC.

Intervención educativa en pacientes con demencia e impacto en la calidad de vida de cuidadores / Educational intervention in patients with dementia and Impact on the caregiver quality of life

Introducción: El deterioro de la memoria que se asocia con la edad (DEMAE) es un trastorno relativamente benigno, que puede ser bastante común por el hecho de vincularse con el envejecimiento normal y el evidente incremento de la expectativa de vida en el ámbito mundial y nacional, con una mayor población de personas de la tercera edad y el consecuente aumento de enfermedades crónicas, propias de este segmento poblacional. Objetivo: Evaluar el resultado de una intervención educativa en pacientes con demencia ligera y su impacto en la calidad de vida del cuidador. Métodos: Se realizó una intervención educativa a pacientes con diagnóstico de demencia ligera en el municipio Madruga de septiembre de 2015 a febrero de 2016. El universo se conformó por 110 ancianos. La muestra se integró por 16 pacientes que se asignaron al grupo de estudio conjuntamente con los 16 cuidadores y 16 al grupo de control mediante asignación aleatoria. Se controlaron variables como deterioro cognitivo, síntomas asociados y calidad de vida. La información se recogió a partir de entrevistas, encuestas a cuidadores y Minimental Test que se aplicaron a los pacientes antes y después de la intervención. Se utilizaron técnicas como la terapia de reminiscencia, repaso de la vida y reeducación al paciente. Resultados: Mejoró el negativismo, el miedo, la distractilidad y la comunicación de los pacientes. Se constató enlentecimiento del deterioro cognitivo en el grupo de estudio y progresó en el grupo control. Mejoró la calidad de vida del cuidador. Conclusiones: Mejoraron los síntomas conductuales, se retardó la evolución de la enfermedad en el grupo de estudio y progresó en el grupo control. Mejoró la calidad de vida del cuidador)AU)

Introduction: Age-related memory impairment is a relatively benign disorder, which can be quite common due to the fact of being associated with normal aging and the evident increase in life expectancy worldwide and nationally, with a greater population of people belonging to the third age and the consequent increase of chronic diseases, characteristic of this population group. Objective: To evaluate the result of an educational intervention in patients with mild dementia and its impact on the quality of life of the caregiver. Methods: An educational intervention was carried out with patients diagnosed with mild dementia in Madruga Municipality from September 2015 to February 2016. The study population was made up by 110 elderly people. The sample was integrated by 16 patients who were assigned to the study group together with the 16 caregivers and 16 assigned to the control group by random assignment. Variables such as cognitive impairment, associated symptoms and quality of life were controlled. The information was collected from interviews, caregiver surveys and the Minimental Test that were applied to patients before and after the intervention. Techniques such as reminiscence therapy, life review and patient reeducation were used. Results: Negativism, fear, distractibility and communication of patients improved. Deceleration of cognitive impairment was observed in the study group and progressed in the control group. The caregiver quality of life improved. Conclusions: Behavioral symptoms improved, the evolution of the disease was delayed in the study group and progressed in the control group. The caregiver quality of life improved(AU)

Intervención educativa sobre infecciones de trasmisión sexual en adolescentes / Educative intervention about sexually transmitted infections in adolescents

Introducción: los adolescentes son muy inestables en sus relaciones sexuales, de ahí la importancia que conozcan sobre las infecciones de trasmisión sexual a través de una intervención educativa. Objetivo: evaluar una estrategia de Intervención Educativa en adolescentes sobre las infecciones de trasmisión sexual y el VIH/SIDA. Métodos: investigación de desarrollo, estudio de tipo interventivo en adolescentes del Área de Aguacate, Municipio Madruga, durante el año 2013, universo constituido por 91 adolescentes, seleccionándose como muestra por criterio de identificación 26 adolescentes de 8vo grado a los cuales se les aplicó la entrevista semiestructurada y la intervención educativa, se controló la edad de comienzo de las relaciones sexuales, el nivel de conocimiento antes y después de la intervención sobre factores de riesgo, vías de trasmisión sexual, actitud ante el contagio y sobre las infecciones de trasmisión sexual y el VIH/SIDA. Resultados: de la muestra estudiada, 17 adolescentes (65,3 por ciento) mantenían relaciones sexuales, el nivel de conocimiento fue malo antes de la intervención y bueno después en todas las variables medidas: factores de riesgo (53,8 por ciento, y 80,7 por ciento respectivamente), vías de trasmisión sexual (46,1 por ciento y 84,6 por ciento respectivamente), sobre infecciones de trasmisión sexual y el VIH/SIDA (57,6 por ciento y 76,9 por ciento respectivamente). La actitud ante el contagio no le preocupaba a 11 adolescentes (42,3 por ciento) y después de la intervención solo a 1 (3,8 por ciento). Conclusiones: se modificaron los conocimientos de los adolescentes sobre las infecciones de trasmisión sexual y el VIH/SIDA luego de la intervención educativa, evaluándose la misma de satisfactoria(AU)

Introduction: Adolescents are very instable in their sexual relationships, therefore the importance for them to know about sexually transmitted infections by means of an educative intervention. Objective: Assess an educative intervention strategy in adolescents about sexually transmitted infections and HIV/AIDS. Methods: Development research, intervention study in adolescents from the Aguacate area, Madruga Municipality, during the year 2013, universe constituted by 91 adolescents, while the sample chosen by identification criteria was made up by 26 adolescents in eighth grade who were applied the semi-structured interview and the educative intervention. We controlled the age to have started sexual intercourse, the level of knowledge before and after the interventions about risk factors, the sexual transmission ways, attitude to the contagion and about the sexually transmitted infections and HIV/AIDS. Results: 17 adolescents (65.3 percent) of the studied sample maintained sexual intercourse, the level of knowledge was mad before the interventions and good later on and regarding all the variable measured: risk factors (53.8 percent and 80.7 percent, respectively), sexual transmission ways (46.1 percent and 84.6 percent, respectively). The attitude to the contagion was not concerned by 11 adolescents (42.3 percent), and after the intervention just by one (3.8 percent). Conclusions: The adolescents' knowledge about sexually transmitted infections and HIV/IADS was modified after the educative intervention, which was assessed as satisfactory(AU)

Tratamiento del síndrome lumbálgico agudo con auriculoterapia / Acute lumbar syndrome treatment with auriculotherapy

Introducción: en el primer nivel de atención, las lumbalgias representan el 5 por ciento de las consultas. Se utiliza un abordaje racional, siendo tratados con técnicas como la Auriculoterapia. Objetivo: evaluar el uso de la técnica de Auriculoterapia en el tratamiento del síndrome lumbálgico agudo. Métodos: estudio exploratorio, prospectivo, en pacientes con síndrome lumbálgico que asistieron al cuerpo de guardia del Policlínico María Emilia Alfonzo de Madruga, en el período de enero del 2015 a enero del 2016. Universo constituido por 120 pacientes, se seleccionó una muestra no probabilística de tipo intencionada de 60 pacientes diagnosticados por ortopedia, divididos en dos grupos para el tratamiento: grupo I: fisioterapia convencional indicada más medicamentos según la sintomatología del proceso, grupo II: fisioterapia convencional indicada más medicamentos según la sintomatología del proceso más Auriculoterapia según el diagnóstico asiático y la exploración del pabellón auricular. Se utilizó la hoja de cargo y la historia clínica individual, se tuvieron en cuenta variables como edad, sexo, diagnóstico según causa, síntomas asociados y evolución del tratamiento en ambos grupos. Resultados: predominó el grupo de edades de 56 a 65 años para un 37 por ciento, del sexo femenino para un 65 por ciento, diagnóstico de artrosis con 49 pacientes para un 82 por ciento, síntomas asociados de dolor en 60 para un 100 por ciento, con evolución buena en 42 para un 70 por ciento. Conclusiones: los pacientes con síndrome lumbálgico evolucionaron satisfactoriamente con el tratamiento convencional, siendo más efectivo el tratamiento convencional más Auriculoterapia(AU)

Introduction: In the first level of health care, lumbagos represent 5 percent of the consultations. A rational approach is used, being treated with techniques such as auriculotherapy. Objective: Assess the use of the technique auriculotherapy to treat the acute lumbar syndrome. Methods: Exploratory, prospective study on patients with lumbar syndrome who attended the emergency room of María Emilia Alfonzo Polyclinic of Madruga, in the period from January 2015 to January 2016. The universe was made up by 120 patients, we chose a nonprobabilistic and nontargeted sample of 60 patients diagnosed by orthopaedics, divided into two groups for treatment: group I: conventional physiotherapy and medication indicated for the process symptoms; group II: conventional physiotherapy and medication indicated for the process symptoms, and auriculotherapy, for the Asian diagnostic and the auricular pavilion exploration. The chart and person's clinical record were used, and variables such as age, sex, cause diagnosis, associated symptoms and treatment evolution were considered in both groups. Results: The predominating age group was 56-65 years (37 percent), female sex (65 percent), arthrosis diagnosed with 49 patients (82 percent), pain-associated symptoms in 60 (100 percent), with good evolution (70 percent). Conclusions: The patients with lumbar syndrome evolved satisfactorily with the conventional treatment, being more effective the conventional treatment plus auriculotherapy(AU)

A hyperpolarization-activated, cyclic nucleotide-gated, (Ih-like) cationic current and HCN gene expression in renal inner medullary collecting duct cells.

The cation conductancein primary cultures of rat renal inner medullary collecting duct was studied using perforated-patch and conventional whole cell clamp techniques. Hyperpolarizations beyond -60 mV induced a time-dependent inward nonselective cationic current (I(vti)) that resembles the well-known hyperpolarization-activated, cyclic nucleotide-gated I(h) and I(f) currents. I(vti) showed a half-maximal activation around -102 mV with a slope factor of 25 mV. It had a higher conductance (but, at its reversal potential, not a higher permeability) for K(+) than for Na(+) (gK(+)/gNa(+) = 1.5), was modulated by cAMP and blocked by external Cd(2+) (but not Cs(+) or ZD-7288), and potentiated by a high extracellular K(+) concentration. We explored the expression of the I(h) channel genes (HCN1 to -4) by RT-PCR. The presence of transcripts corresponding to the HCN1, -2, and -4 genes was observed in both the cultured cells and kidney inner medulla. Western blot analysis with HCN2 antibody showed labeling of approximately 90- and approximately 120-kDa proteins in samples from inner medulla and cultured cells. Immunocytochemical analysis of cell cultures and inner medulla showed the presence of HCN immunoreactivity partially colocalized with the Na(+)-K(+)-ATPase at the basolateral membrane of collecting duct cells. This is the first evidence of an I(h)-like cationic current and HCN immunoreactivity in either kidney or any other nonexcitable mammalian cells.