Descending serotonergic controls regulate inflammation-induced mechanical sensitivity and methyl-CpG-binding protein 2 phosphorylation in the rat superficial dorsal horn

<p>Abstract</p> <p>Background</p> <p>Regulation of pain states is, in part, dependent upon plastic changes in neurones within the superficial dorsal horn. There is also compelling evidence that pain states are under the control of descending projections from the brainstem. While a number of transcription factors including Methyl-CpG-binding protein 2 (MeCP2), Zif268 and Fos have been implicated in the regulation of dorsal horn neurone sensitization following injury, modulation of their activity by descending controls has not been investigated.</p> <p>Results</p> <p>Here, we describe how descending controls regulate MeCP2 phosphorylation (P-MeCP2), known to relieve transcriptional repression by MeCP2, and Zif268 and Fos expression in the rat superficial dorsal horn, after CFA injection into the hind paw. First, we report that CFA significantly increased P-MeCP2 in Lamina I and II, from 30 min post injection, with a maximum reached after 1 h. The increase in P-MeCP2 paralleled that of Zif268 and Fos, and P-MeCP2 was expressed in large sub-populations of Zif268 and Fos expressing neurones. Serotonergic depletion of the lumbar spinal cord with 5,7 di-hydroxytryptamine creatinine sulphate (5,7-DHT) reduced the inflammation evoked P-MeCP2 in the superficial dorsal horn by 57%, and that of Zif268 and Fos by 37.5% and 30% respectively. Although 5,7-DHT did not change primary thermal hyperalgesia, it significantly attenuated mechanical sensitivity seen in the first 24 h after CFA.</p> <p>Conclusion</p> <p>We conclude that descending serotonergic pathways play a crucial role in regulating gene expression in the dorsal horn and mechanical sensitivity associated with an inflammatory pain state.</p

Cultivo in vitro de láminas de corteza ovárica en porcinos : efecto del suero fetal bovino y de la hormona folículo estimulante sobre la integridad de los folículos preantrales

El cultivo in vitro de láminas de tejido ovárico sería un método eficaz para analizar el potencial daño criogénico sobre los folículos preantrales (FPA). El objetivo fue evaluar el efecto de distintos porcentajes de suero fetal bovino (SFB) en el medio de cultivo en presencia o ausencia de hormona folículo estimulante (FSH), sobre la morfología y morfometría de los FPA porcinos en láminas de corteza ovárica. Se utilizaron ovarios de faena (n=4). Las muestras se cultivaron 24 h a 39 °C, en cámara húmeda (5% de CO2), en medio D-MEM, ITS (insulina 1mg/L, transferrina 0,55 mg/mL, selenio 0,5 μg/mL), glutamina 2 mM, gentamicina y anfotericina B 10 μg/mL, con SFB (0 %, 5%, 10% y 20%), en ausencia o presencia (50 ng/mL) de FSH. Se evaluaron morfológicamente cortes teñidos con Hematoxilina-Eosina por microscopía de campo claro (x400). Se analizó la morfométria sobre fotos digitales obtenidas con cámara LEICA DCC-380X® (programa de captura LAS LEICA Inc ®). Se determinó el área nuclear (AN), citoplasmática (AC) y total de cada ovocito con el software Leica Qwin Plus® 3.1. Se calculó la relación de área núcleo/citoplasma (AN/AC). Se analizaron 505 FPA (Test de Friedman p&lt;0,05). Los medios de cultivo con 0% y 5% de SFB sin FSH produjeron una reducción significativa en el porcentaje de FPA normales con respecto al control y a los restantes tratamientos. Los FPA primordiales presentaron una reducción significativa del AN y AN/AC. En presencia de FSH combinada con 20% de SFB, se observó una reducción significativa del porcentaje de FPA morfológicamente normales con respecto al control y los demás tratamientos. Se sugeriría la suplementación del medio de cultivo con 10% de SFBFil: Gabriel, P.. Buenos Aires (Argentina). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Fil: Cisale, H.. Buenos Aires (Argentina). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Fil: Fischman, M.. Buenos Aires (Argentina). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Fil: Fratto, M.. Buenos Aires (Argentina). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).Fil: Lombardo, D.. Buenos Aires (Argentina). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)

Evaluation of the capillary electrophoresis method for measurement of immunoglobulin concentration in ewe colostrum

ABSTRACT Capillary electrophoresis (CE) is a technique routinely used in clinical laboratories that allows the separation and quantification of blood serum proteins in a rapid, precise, accurate, and inexpensive manner. Recently, CE has been proposed to separate and measure colostral proteins, but an evaluation of the agreement between CE and radial immunodiffusion (RID) method, currently used to quantify IgG in colostrum, is still lacking. The purpose of this study was to test the ability of a CE instrument, normally used in blood serum protein analysis, to realize the correct quantification of total Ig concentration in ewe colostrum, using RID assay as reference. Colostrum samples (n = 68) were collected from 35 multiparous Sarda ewes at first milking (n = 33) and at 24 h postpartum (n = 35). The mean ± standard deviation of IgG concentration measured by RID and whey colostrum total Ig concentration measured by CE were 54.76 ± 41.82 g/L and 54.70 ± 41.43 g/L, respectively. Lin's concordance correlation coefficient (r = 0.993; 95% confidence interval=0.989 to 0.996) and linear regression analysis results (RID = 1.0022CE − 0.063; R 2 = 0.986) showed an excellent agreement between these 2 methods. Bland-Altman analysis confirmed that CE method can be a suitable alternative to RID: the mean of the differences between CE and RID was −0.055 ± 4.95 g/L (95% confidence interval=−1.25 to 1.14 g/L) and the agreement limits were −9.75 to 9.60 g/L (low limit 95% confidence interval=−11.82 to −7.68 g/L; high limit 95% confidence interval=7.57 to 11.72 g/L). In conclusion, the current study indicates that CE method may be a reliable tool for the quantification of the total Ig concentration in ewe colostrum

New molecular targets in bone metastases

Bone metastases have a major impact on morbidity and on mortality in cancer patients. Despite its clinical relevance, metastasis remains the most poorly elucidated aspect of carcinogenesis. The biological mechanisms leading to bone metastasis establishment have been referred as " vicious circle," a complex network between cancer cells and the bone microenvironment. This review is aimed to underline the new molecular targets in bone metastases management other than bisphosphonates. Different pathways or molecules such as RANK/RANKL/OPG, cathepsin K, endothelin-1, Wnt/DKK1, Src have recently emerged as potential targets and nowadays preclinical and clinical trials are underway. The results from those in the advanced clinical phases are encouraging and underlined the need to design large randomised clinical trials to validate these results in the next future.Targeting the bone by preventing skeletal related events (SREs) and bone metastases has major clinical impact in improving survival in bone metastatic patients and in preventing disease relapse in adjuvant setting. © 2010 Elsevier Ltd

Prognostic role of human equilibrative transporter 1 (hENT1) in patients with resected gastric cancer

Nucleoside transporter proteins are specialized proteins that mediate the transport of nucleosides and nucleoside analog drugs across the plasma membrane. The human equilibrative nucleoside transporter 1 (hENT1) is a member of these proteins and mediates cellular entry of gemcitabine, cytarabine, and fludarabine. The hENT1 expression has been demonstrated to be related with prognosis and activity of gemcitabine-based therapy in breast, ampullary, lung, and pancreatic cancer. We investigated the immunohistochemical expression of hENT in tumor samples from III patients with resected gastric adenocarcinoma, correlating these data with clinical parameters and disease outcomes. None of the patients received chemotherapy or radiation therapy before or after surgery as a part of an adjuvant or neoadjuvant program. On univariate survival analysis, the hENT1 expression was associated with overall survival (OS) and disease free survival (DFS). Specifically, those patients with overexpression of hENT1 showed a shorter OS ( P=0.021) and a shorter DFS ( P=0.033). Considering only the node positive patients, higher hENT levels were associated with significantly shorter median DFS (21.7 months; 95% CI 11.1-32.4) compared with patients with low expression of hENT1. The hENT1 expression was defined, in the lymph-node positive patients, as an independent prognostic factor ( P=0.019). Furthermore, considering only patients with diffuse or mixed tumors and lymph-node positive, the expression of hENT1 was strongly related with DFS and OS. Immunohistochemistry for the hENT1 protein carries prognostic information in patients with resected gastric cancer and holds promise as a predictive factor in chemotherapy decisions

Cognitive Changes and Quality of Life in Neurocysticercosis: A Longitudinal Study

Neurocysticercosis (NCC) is one of the most common parasitic infections of the central nervous system. Cognitive changes have been frequently reported with this disease but have not been well studied. Our study team recruited a group of new onset NCC cases and a matched set of healthy neighborhood controls and new onset epilepsy controls in Lima, Peru for this study. A neuropsychological battery was administered at baseline and at 6 months to all groups. Brain MRI studies were also obtained on NCC cases at baseline and at 6 months. Newly diagnosed patients with NCC had mild cognitive deficits and more marked decreases in quality of life at baseline compared with controls. Improvements were found in both cognitive status and quality of life in patients with NCC after treatment. This study is the first to assess cognitive status and quality of life longitudinally in patients with NCC and provides new data on an important clinical morbidity outcome