Measurement of the top quark mass in the dilepton channel using mT2 at CDF

We present measurements of the top quark mass using the \mT2, a variable related to the transverse mass in events with two missing particles. We use the template method applied to t\tbar dilepton events produced in p\pbar collisions at Fermilab's Tevatron and collected by the CDF detector. From a data sample corresponding to an integrated luminosity of 3.4 \invfb, we select 236 t\tbar candidate events. Using the \mT2 distribution, we measure the top quark mass to be M_{Top} = 168.0^{+4.8}_{-4.0} $\pm$ {2.9} GeV/c^{2}. By combining the \mT2 with the reconstructed top mass distributions based on a neutrino weighting method, we measure M_{top}=169.3 $\pm$ 2.7 $\pm$ 3.2 GeV/c^{2}. This is the first application of the \mT2 variable in a mass measurement at a hadron collider.

Measurement of the top quark mass in the dilepton channel using mT2 at CDF

We present measurements of the top quark mass using the \mT2, a variable related to the transverse mass in events with two missing particles. We use the template method applied to t\tbar dilepton events produced in p\pbar collisions at Fermilab's Tevatron and collected by the CDF detector. From a data sample corresponding to an integrated luminosity of 3.4 \invfb, we select 236 t\tbar candidate events. Using the \mT2 distribution, we measure the top quark mass to be M_{Top} = 168.0^{+4.8}_{-4.0} $\pm$ {2.9} GeV/c^{2}. By combining the \mT2 with the reconstructed top mass distributions based on a neutrino weighting method, we measure M_{top}=169.3 $\pm$ 2.7 $\pm$ 3.2 GeV/c^{2}. This is the first application of the \mT2 variable in a mass measurement at a hadron collider.

Melatonin acts through MT1/MT2 receptors to activate hypothalamic Akt and suppress hepatic gluconeogenesis in rats

Melatonin can contribute to glucose homeostasis either by decreasing gluconeogenesis or by counteracting insulin resistance in distinct models of obesity. However, the precise mechanism through which melatonin controls glucose homeostasis is not completely understood. Male Wistar rats were administered an intracerebroventricular (icv) injection of melatonin and one of following: an icv injection of a phosphatidylinositol 3-kinase (PI3K) inhibitor, an icv injection of a melatonin receptor (MT) antagonist, or an intraperitoneal (ip) injection of a muscarinic receptor antagonist. Anesthetized rats were subjected to pyruvate tolerance test to estimate in vivo glucose clearance after pyruvate load and in situ liver perfusion to assess hepatic gluconeogenesis. The hypothalamus was removed to determine Akt phosphorylation. Melatonin injections in the central nervous system suppressed hepatic gluconeogenesis and increased hypothalamic Akt phosphorylation. These effects of melatonin were suppressed either by icv injections of PI3K inhibitors and MT antagonists and by ip injection of a muscarinic receptor antagonist. We conclude that melatonin activates hypothalamus-liver communication that may contribute to circadian adjustments of gluconeogenesis. These data further suggest a physiopathological relationship between the circadian disruptions in metabolism and reduced levels of melatonin found in type 2 diabetes patients.

Effect of Melatonin and Analogues on Corneal Wound Healing: Involvement of Mt2 Melatonin Receptor

Purpose: We have investigated the effect of melatonin and its analogues on rabbit corneal epithelial wound healing. Methods: New Zealand rabbits were anaesthetised and wounds were made by placing Whatman paper discs soaked in n-heptanol on the cornea. Melatonin and analogues (all 10 nmol) were instilled. Wound diameter was measured every 2 hours by means of fluorescein application with a Topcon SL-8Z slit lamp. Melatonin antagonists (all 10 nmol) were applied 2 hours before the application of the n-heptanol-soaked disc and then every 6 hours together with melatonin. To confirm the presence of MT2 receptors in corneal epithelial cells immunohistochemistry, Western blot and RT-PCR assays in native tissue and in rabbit corneal epithelial cells were performed. The tear components were extracted then processed by HPLC to quantify melatonin in tears. Results: Migration assays revealed that melatonin and particularly the treatment with the MT2 agonist IIK7, accelerated the rate of healing (p < 0.001). The application of the non-selective melatonin receptor antagonist luzindole and the MT2 antagonist DH97 (but not prazosin), prevented the effect of melatonin on wound healing (both p < 0.001). Immunohistochemistry, Western blot and RT-PCR assays showed the presence of MT2 melatonin receptor in corneal epithelial cells. In addition, we have identified melatonin in tears and determined its daily variations. Conclusions: These data suggest that MT2 receptors are implicated in the effect of melatonin on corneal wound healing regulating migration rate. This suggests the potential use of melatonin and its analogues to enhance epithelial wound healing in ocular surface disease.

Upregulated MT1-MMP/TIMP-2 axis in the TSU-Pr1-B1/B2 model of metastatic progression in transitional cell carcinoma of the bladder

Muscle invasive transitional cell carcinoma (TCC) of the bladder is associated with a high frequency of metastasis, resulting in poor prognosis for patients presenting with this disease. Models that capture and demonstrate step-wise enhancement of elements of the human metastatic cascade on a similar genetic background are useful research tools. We have utilized the transitional cell carcinoma cell line TSU-Pr1 to develop an in vivo experimental model of bladder TCC metastasis. TSU-Pr1 cells were inoculated into the left cardiac ventricle of SCID mice and the development of bone metastases was monitored using high resolution X-ray. Tumor tissue from a single bone lesion was excised and cultured in vitro to generate the TSU-Pr1-B1 subline. This cycle was repeated with the TSU-Pr1-B1 cells to generate the successive subline TSU-Pr1-B2. DNA profiling and karyotype analysis confirmed the genetic relationship of these three cell lines. In vitro, the growth rate of these cell lines was not significantly different. However, following intracardiac inoculation TSU-Pr1, TSU-Pr1-B1 and TSU-Pr1-B2 exhibited increasing metastatic potential with a concomitant decrease in time to the onset of radiologically detectable metastatic bone lesions. Significant elevations in the levels of mRNA expression of the matrix metalloproteases (MMPs) membrane type 1-MMP (MT1-MMP), MT2-MMP and MMP-9, and their inhibitor, tissue inhibitor of metalloprotease-2 (TIMP-2), across the progressively metastatic cell lines, were detected by quantitative PCR. Given the role of MT1-MMP and TIMP-2 in MMP-2 activation, and the upregulation of MMP-9, these data suggest an important role for matrix remodeling, particularly basement membrane, in this progression. The TSU-Pr1-B1/B2 model holds promise for further identification of important molecules.

Upregulation of matrix metalloproteinases (MMPs) in breast cancer xenografts : a major induction of stromal MMP-13

In human breast cancer (HBC), as with many carcinoma systems, most matrix metalloproteinases (MMPs) are largely expressed by the stromal cells, whereas the tumour cells are relatively silent in MMP expression. To determine the tissue source of the most relevant MMPs, we xenografted HBC cell lines and HBC tissues into the mammary fat pad (MFP) or bone of immunocompromised mice and measured the expression of human and mouse MMP-2, -9, -11, -13, membrane-type-1 MMP (MT1-MMP), MT2-MMP and MT3-MMP by species-specific real-time quantitative RT-PCR. Our data confirm a stromal origin for most tumour-associated MMPs and indicate marked and consistent upregulation of stromal (mouse) MMP-13 and MT1-MMP in all xenografts studied, irrespective of implantation in the MFP or bone environments. In addition, we show increased expression of both human MMP-13 and human MT1-MMP by the MDA-MB-231 tumour cells grown in the MFP compared to in vitro production. MMP protein and activity data confirm the upregulation of MMP mRNA production and indicate an increase in the activated MMP-2 species as a result of tumour implantation. These data directly demonstrate tumour induction of MMP production by stromal cells in both the MFP and bone environments. These xenografts are a valuable means for examining in vivo production of MMPs and suggest that MMP-13 and MT1-MMP will be relevant targets for inhibiting breast cancer progression.

Correlation of tumor- and stromal-derived MT1-MMP expression with progression of human ovarian tumors in SCID mice

Human ovarian carcinoma samples were orthotopically implanted into SCID mice to investigate the contribution of matrix metalloproteases (MMPs) to the spread of ovarian tumors. Mice were inoculated with patient tumor samples, and developed ovarian tumors over a 16-week period with metastasis occurring in some mice. Species-specific quantitative RT-PCR was used to identify the source of tumor-associated MMPs. Membrane-type (MT)1-MMP mRNA was significantly increased in high-grade tumors, tumors with evidence of serosal involvement, and tumors in which distant metastases were detected. The increase in MT1-MMP expression was predominantly from the human tumor cells, with a minor contribution from the mouse ovarian stroma. Neither human nor mouse MT2-MMP were correlated with tumor progression and MT3-MMP levels were negligible. While tumor cells did not produce significant amounts of MMP-2 or MMP-9, the presence of tumor was associated with increased levels of MMP-2 expression by mouse ovarian stroma. Stromal-derived MT1-MMP was greater in large tumors and was associated with stromal MMP-2 expression but neither was significantly linked with metastasis. These studies indicate that tumor-derived MT1-MMP, more so than other gelatinolytic MMPs, is strongly linked to aggressive tumor behavior. This orthotopic model of human ovarian carcinoma is appropriate for studying ovarian tumor progression, and will be valuable in the further investigation of the metastatic process.

Implication of collagen type I-induced membrane-type 1-matrix metalloproteinase expression and matrix metalloproteinase-2 activation in the metastatic progression of breast carcinoma

We have previously demonstrated that fibroblasts and invasive human breast carcinoma (HBC) cells specifically activate matrix metalloproteinase- 2 (MMP-2) when cultured on 3-dimensional gels of type I collagen but not a range of other substrates. We show here the constitutive expression of membrane-type 1 (MT1)-MMP in both fibroblasts, and invasive HBC cell lines, that have fibroblastic attributes presumably acquired through an epithelial- to-mesenchymal transition (EMT). Treatment with collagen type I increased the steady-state MT1-MMP mRNA levels in these cells but did not induce either MT1-MMP expression or MMP-2 activation in noninvasive breast carcinoma cell lines, which retain epithelial features. Basal MT3-MMP mRNA expression had a pattern similar to that of MT1-MMP but was not up-regulated by collagen. MT4- MMP mRNA was seen in both invasive and noninvasive HBC cell lines and was also not collagen-regulated, and MT2-MMP mRNA was not detected in any of the HBC cell lines tested. These data support a role for MT1-MMP in the collagen- induced MMP-2-activation seen in these cells. In situ hybridization analysis of archival breast cancer specimens revealed a close parallel in expression of both collagen type I and MT1-MMP mRNA in peritumoral fibroblasts, which was correlated with aggressiveness of the lesion. Relatively high levels of expression of both mRNA species were seen in fibroblasts close to invasive tumor nests and, although only focally, in certain areas close to preinvasive tumors. These foci may represent hot spots for local degradation and invasive progression. Collectively, these results implicate MT1-MMP in collagen- stimulated MMP-2 activation and suggest that this mechanism may be employed in vivo by both tumor-associated fibroblasts and EMT-derived carcinoma cells to facilitate increased invasion and/or metastasis.

Efecto de tres presentaciones de pienso sobre la productividad en conejos de engorde

El presente trabajo experimental se realizó, con el objetivo de estudiar el efecto de presentación del alimento, sobre los rendimientos productivos en conejos de engorde. Se aplica con tres tratamientos. A los animales del grupo control (Tl) se les suministró un pienso comercial en forma de harina. En el segundo tratamiento (T2) se suministró el mismo pienso en forma de amasijo (76% harina y 24% agua) y en el tercer tratamiento (T3) los animales consumieron el mismo pienso comercial pero en forma de gránulos o pellet. Se utilizaron 45 gazapos (de ambos sexos), de 42 días de edad de la raza Neozelandés blanco (NZB), con un peso vivo inicial promedio de 0.877 Kg. y se llevaron a los 77 días de edad con un promedio final de 1.849 Kg. de peso vivo. Para la evaluación estadística se empleó un diseño completamente aleatorio (DCA) con tres tratamientos y cinco repeticiones por tratamiento. Cada unidad experimental (jaulas de 0.40 mt2) estaba constituida por tres gazapos distribuidos aleatoriamente a los que se les registraron diariamente el consumo de alimento y semanalmente el aumento de peso vivo. Los parámetros (ANDEVA) fueron la promedio de alimento cada tratamiento. analizados estadísticamente por medio de ganancia media diaria (Kg/día), el consumo (Kg. MS/día) y el índice de conversión por todas las variables estudiadas se vieron afectadas por la forma de presentación del pienso y se registraron diferencias significativas entre los tratamientos, resultando que la ganancia media diaria (kg./día), fue similar en los tratamientos T1 y T2, pienso en forma de harina y pienso en forma de amasijo; pero si hubo diferencias significativas (P < 0.01) en relación al tratamiento T3, pienso en forma de gránulo (I, 0.02525; II, 0.02406 y III, 0.03395) respectivamente. Nuestros resultados demuestran que el consumo de pienso (kg. MS/día) difiere significativamente (P < 0.01) con un mayor consumo de pienso granulado (I, 0.06333; II, 0.07478 y III, 0.08854) respectivamente. A través del procedimiento de separación de medias (D.M.S.) se encontró que los índices de conversión para los alimentos en forma de harina y gránulo o pellet fueron similares, pero fue mayor el valor presentado por el amasijo (I, 2,51; II, 3.10 y III, 2.60) respectivamente. Durante el ensayo los animales del tratamiento tres (pienso granulado) fueron los primeros en alcanzar el peso comercial (2,072 kg.) a los 35 días del engorde, en tanto en los tratamientos I y II los pesos finales registrados fueron 1,753 y 1.721 kg de peso vivo respectivamente. Luego de analizados los resultados obtenidos se pudo detectar diferencias significativas debido a un mayor consumo y crecimiento en los animales que consumieron pienso granulado.

Valor nutritivo del pasto pangola (Digitaria decumbens stent.) vr. transvala a distintos niveles de fertilización y diversas edades de corte

Este estudio Se desarrolló en el periodo de julio a diciembre de 1990 en el Centro de Investigación Zootecnia “La Polvosa” de la UCA (Universidad Centroamericana), situada en el km 23 112 de la carretera nueva a León, depto. de Managua. El centro se ubica a una elevación de 40 msnrn a 1 2 grados 12 minutos, Latitud norte y 86 grados 22 minutos, longitud oeste registra temperaturas promedio anuales de 32 grados centígrados los y unos 8OO mm de precipitación media por año, por lo cual, es posible calificar su zona agroecológica como de trópico seco se evaluó la respuesta en términos de valor nutritivo del pasto Pangola (Digitaria decumbens Stent.) vr. Transvala, utilizándose un diseño experimental en bloques completos al azar (BCA) con arreglo factorial, se estudió el efecto de dos factores (Niveles de fertilizantes-edades de cortes) con cuatro repeticiones, formándose un total de 16 tratamientos El ensayo contó con un área experimental total de 357.75 mt2 para la toma de muestras, se empleó el método del metro cuadrado realizándose en las parcelas un muestreo sin reemplazo para cada frecuencia en estudio. Previo al momento experimental, se efectúo una poda de control procediéndose después a aplicar de una sola vez los respectivos niveles de fertizante. El estudio estadístico contempló el uso del análisis de varianza (ANDEVA), se hizo una separción de medias por Tukey, y se midió la influencia porcentual (Individual y asociada) de 4 niveles de fertilizante (0, 50,100 y150 kg urea (46%N2) / mz) y 4 edades de corte (15, 30.. 45y 60 días), sobre nueve variables o componentes bromatológicos (MS, PB,FC, ELN, EE, EB (Kcal/ 100g), Cz, Ca y P). Todos los componentes bromatológicos evaluados excepto el EE.. presentaron una variación significativa (P<0.05), bajo el efecto de los factores en estudio. Los niveles de fertilizante influyeron ios valores de mayor mérito con la dosis •150 kg urea / rnz, observando la PB.. 9.76% ;el ELN, 56.54 y la EB,190.97 Kcal ,' 100 g. En tanto que los 50 kg1mz mostraron !os mayores índices en FC (28.88), EE (2.75) y Cz. (11.:39) las parcelas no fertilizadas brindaron los porcentajes más altos de MS (23.78) y Ca (0.60),y el nivel 100 kg/ mz El mejor en P (0.39) y el más bajo en FC (26.61). A su vez con O kg / mz_. Observaron sus níveles minimos la PB (6.91), el ELN (53.58). el valor energético (175.45 Kcal/ 100 g) y el P (O.28), y con 100 kg/ mz los menores en MS y Cz (25.08 y 10.38 respectivamente), a. los 150kg/mz el EE (2.29) y el Ca (0.49), rindieron sus índices más bajos. En cuanto a la frecuencia de corte, la edad 15 días presentó los máximos contenidos en PH (11.95), ELN (56.45),EB (202.40 Kcal/ 100g), EE (2.65) y el mínimo en FC (27.09). Observando el corte a los 60 días, las mayores proporciones en MS (30.91) Cz (12.43) )'Ca (0.58),en tanto que con 30 y 45 días se dieron los más altos valores de P (0.34) FC (28.55) respectivamente los índices más bajos de PB (6.48),ELN (52.63), Ca (0.47)y P (0.30), se obtuvieron a los 45 días de madurez mientras que la MS (22.88) y la Cz (9.56),alcanzaron sus valores más pequeños con 15 días de corte a los 60 días, la EB (175. 42 Kcal ./ 100g) y el EE (2.20) respondieron con sus por cientos más bajos. Para los tratamientos resultantes de la combinación de un nivel de fertilizante y una edad de corte, la interacción 50 kg/mz-60 días reportó el mayor contenido en MS (32.72), sobresaliendo el tratamiento 150 kg / mz-15 días al presentar los valores más altos en EB (214.39 Kcal/100g), PB (14.30) y ELN (59.02), aunque la combinación 50 kg/mz-30 días brindó igual valor en ELN y el máximo en Cz (13.10) con aplicaciones de 100 kg/mz y frecuencia de 15 días se obtuvieron los menores índices en FC (24.82) y Cz (9.37). Consiguiéndose en O kg / mz-15 días el valor más alto en Ca (0.76) y el menor en P (0.22), observando la MS su índice más bajo (18.37) con el tratamiento 100 kg / mz-15 días. Finalmente, en las interacciones que incluyen la frecuencia 45 días, se encontró que las parcelas no fertilizadas presentaron los menores contenidos en PB (5.02), ELN (48.95) y EB (165.44 Kcal/ 1oo g). en cambio al aplicar 100 kg /rnz, se obtuvieron (para la misma interacción) los valores más bajos en EE (1.72), Ca (0.43) y el mayor en P (0.44),en tanto los niveles 50 y 150 kg / mz influyeron los índices más altos en EE (3.85) y FC (30.75) respectivamente.

Cloning and characterization of a new multi-stress inducible metallothionein gene in Tetrahymena pyriformis

A new multi-stress-inducible metallothionein (MT) gene isoform has been cloned and characterized from the ciliate Tetrahymena pyriformis. Both the 5'- and 3'-UT regions of the Tp-MT2 gene are very different from the previously reported Tp-MT1 isoform in this organism and from other described MT genes in Tetrahymena pigmentosa and Tetrahymena thermophila. The putative protein sequence of Tp-MT2 contains cysteine clusters with characteristics of the typical Tetrahymena Cd-inducible MT genes. However, the sequence has a special feature of four intragenic tandem repeats within its first half, with a conserved structural pattern x(5/8)CCCx(6)CCx(6)CxCxNCxCCK. To investigate the transcriptional activities of both Tp-MT2 and Tp-MT1 genes toward heavy metals (Cd, Hg, Cu, Zn) and H2O2, the mRNA levels of these two isoforms were evaluated by means of real-time quantitative PCR. Results showed that Tp-MT2 had a higher basal expression level than Tp-MT1 and both genes were induced by Cd, Hg, Cu, and Zn ions after short exposure (I h), although to different extents. Cd was the most effective metal inducer of both two isoforms, but the relative expression level of Tp-MT2 was much lower than that of Tp-MT1. Different expression patterns were also shown between the two genes when treated with Cd over a period of 24 h. We suggest that TpMT-1 plays the role of a multi-inducible stress gene, while TpMT-2 may have a more specific function in basal metal homeostasis although it may have undergone a functional differentiation process. The putative functional significance and evolutionary mode of the TpMT-2 isoform are discussed. (c) 2006 Elsevier GmbH. All rights reserved.

Os recetores da melatonina como novos alvos de fármacos antidepressivos

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Ligand arsenic complexation and immunoperoxidase detection of metallothionein in the earthworm Lumbricus rubellus inhabiting arsenic-rich soil

Although earthworms have been found to inhabit arsenic-rich soils in the U.K., the mode of arsenic detoxification is currently unknown. Biochemical analyses and subcellular localization studies have indicated that As3+-thiol complexes may be involved; however, it is not known whether arsenic is capable of inducing the expression of metallothionein (MT) in earthworms. The specific aims of this paper were (a) to detect and gain an atomic characterization of ligand complexing by X-ray absorption spectrometry (XAS), and (b) to employ a polyclonal antibody raised against an earthworm MT isoform (w-MT2) to detect and localize the metalloprotein by immunoperoxidase histochemistry in the tissues of earthworms sampled from arsenic-rich soil. Data suggested that the proportion of arsenate to sulfur-bound species varies within specific earthworm tissues. Although some arsenic appeared to be in the form of arsenobetaine, the arsenic within the chlorogogenous tissue was predominantly coordinated with S in the form of -SH groups. This suggests the presence of an As::MT complex. Indeed, MT was detectable with a distinctly localized tissue and cellular distribution. While MT was not detectable in the surface epithelium or in the body wall musculature, immunoperoxidase histochemistry identified the presence of MT in chloragocytes around blood vessels, within the typhlosolar fold, and in the peri-intestinal region. Focal immunostaining was also detectable in a cohort of cells in the intestinal wall. The results of this study support the hypothesis that arsenic induces MT expression and is sequestered by the metalloprotein in certain target cells and tissues.

Variants in MTNR1B influence fasting glucose levels.

To identify previously unknown genetic loci associated with fasting glucose concentrations, we examined the leading association signals in ten genome-wide association scans involving a total of 36,610 individuals of European descent. Variants in the gene encoding melatonin receptor 1B (MTNR1B) were consistently associated with fasting glucose across all ten studies. The strongest signal was observed at rs10830963, where each G allele (frequency 0.30 in HapMap CEU) was associated with an increase of 0.07 (95% CI = 0.06-0.08) mmol/l in fasting glucose levels (P = 3.2 x 10(-50)) and reduced beta-cell function as measured by homeostasis model assessment (HOMA-B, P = 1.1 x 10(-15)). The same allele was associated with an increased risk of type 2 diabetes (odds ratio = 1.09 (1.05-1.12), per G allele P = 3.3 x 10(-7)) in a meta-analysis of 13 case-control studies totaling 18,236 cases and 64,453 controls. Our analyses also confirm previous associations of fasting glucose with variants at the G6PC2 (rs560887, P = 1.1 x 10(-57)) and GCK (rs4607517, P = 1.0 x 10(-25)) loci.