Biopsia prostética randomizada: ¿influye el número de muestras y el valor del PSA para la detección del cáncer prostético? / Randomized prosthetic biopsy: does the number of samples and the value of PSA influence the detection of prostate cancer?

Objetivo: Determinar si la realización de un mayor número de muestras según el valor del antígeno prostático específico (PSA) incrementa la detección del cáncer prostático (CaP). Materiales y métodos: Se estudió transversalmente a 994 pacientes sometidos a una biopsia prostática transrrectal ecodirigida randomizada (BPTE), con sospecha de CaP. Los casos fueron divididos en dos grupos: A (esquema de 8 muestras y ampliado) con protocolo normal (n = 819) y B (esquema de 12 muestras o más) con protocolo extendido (n = 175). Estos se subdividieron de acuerdo con el valor del PSA en tres niveles (< 3,9ng/ml, 4-9,9 ng/ml y > 10ng/ml) y se evaluó la tasa de detección de CaP en cada subgrupo. Los datos clínicos fueron analizados con las pruebas T de student, chi-cuadrado y regresión logística, tomando como estadísticamente significativo un valor inferior a 0,05. Resultados: La tasa de detección de CaP en el grupo A fue mayor que la del B: 43,71%vs. 34,29%; p = 0,022. Al analizar los resultados, teniendo en cuenta la subdivisión de los valores de PSA, no encontramos diferencias estadísticamente significativas para la detección de malignidad entre los dos grupos. Conclusión: Aumentar el número de muestras en la BPTE no incrementó la tasa de detección de CaP de forma independiente ni en cada subgrupo de PSA, así como tampoco en los subgrupos de volumen prostático. Por otro lado, a mayor edad y PSA, hubo más detecciones de CaP.

Objective: To determine whether obtaining a larger number of core specimens, depending on the PSA value, increases the detection of prostate cancer. Materials and methods: A cross-sectional study was conducted on 994 patients with suspected prostate cancer who underwent transrectal ultrasound-guided prostate biopsy. The patients weredivided intogroup Awith a standard protocol (8 core specimens or more scheme) and group B with an extended protocol (12 core specimens or more scheme), and subdivided according to the PSA values into three groups (<3.9ng/ml, 4 to 9.9ng/ml, and > 10ng/ml). The prostate cancer detection rate was evaluated in each subgroup. Clinical data was analysed using the Student t and chisquared tests and logistic regression analysis, taking a P< 0.05 as statistically significant. Results: The detection rate of prostate cancer in the group A was higher than group B: 43.71% vs. 34.29%; P = 0.022. When analysing the results, taking into account the sub-division of PSA results, no significant statistical differences were found in the detection of malignancy in the two groups of patients evaluated. Conclusión: Increasing the number of core specimens in transrectal ultrasound-guided prostate biopsy, does not increase the overall detection rate of prostate cancer in either of the PSA sub-groups, or in either of the of prostate volume sub-groups. On the other hand, the older the patient and the higher the PSA value, the greater the detection of prostate cancer.

Beta-defensin-2 genomic copy number variation and chronic periodontitis.

Chronic periodontitis (ChP) is a multifactorial disease influenced by microbial and host genetic variability; however, the role of beta-defensin-2 genomic (DEFB4) copy number (CN) variation (V) in ChP remains unknown. The association of the occurrence and severity of ChP and DEFB4 CNV was analyzed. Our study included 227 unrelated Caucasians, that is, 136 ChP patients (combined ChP) and 91 control individuals. The combined ChP group was subdivided into the severe ChP and slight-to-moderate ChP subgroups. To determine DEFB4 CNV, we isolated genomic DNA samples and analyzed them by relative quantitation using the comparative CT method. The serum beta-defensin-2 (hBD-2) level was determined via ELISA. The distribution pattern and mean DEFB4 CN did not differ significantly in combined ChP cases vs. the controls; however, the mean DEFB4 CN in the severe ChP group differed significantly from those for the control and slight-to-moderate ChP groups. Low DEFB4 CN increased the risk of severe ChP by about 3-fold. DEFB4 CN was inversely associated with average attachment loss. Mean serum hBD-2 levels were highest in the controls, followed by the slight-to-moderate ChP group and the severe ChP group. The results suggested an association between decreased DEFB4 CN and serum hBD-2 levels and periodontitis severity.

Histamine modulates γδ-T lymphocyte migration and cytotoxicity, via Gi and Gs protein-coupled signalling pathways.

BACKGROUND AND PURPOSE: The biogenic amine, histamine plays a pathophysiological regulatory role in cellular processes of a variety of immune cells. This work analyses the actions of histamine on γδ-T lymphocytes, isolated from human peripheral blood, which are critically involved in immunological surveillance of tumours. EXPERIMENTAL APPROACH: We have analysed effects of histamine on the intracellular calcium, actin reorganization, migratory response and the interaction of human γδ T cells with tumour cells such as the A2058 human melanoma cell line, the human Burkitt's Non-Hodgkin lymphoma cell line Raji, the T-lymphoblastic lymphoma cell line Jurkat and the natural killer cell-sensitive erythroleukaemia cell line, K562. KEY RESULTS: γδ T lymphocytes express mRNA for different histamine receptor subtypes. In human peripheral blood γδ T cells, histamine stimulated Pertussis toxin-sensitive intracellular calcium increase, actin polymerization and chemotaxis. However, histamine inhibited the spontaneous cytolytic activity of γδ T cells towards several tumour cell lines in a cholera toxin-sensitive manner. A histamine H(4) receptor antagonist abolished the histamine induced γδ T cell migratory response. A histamine H(2) receptor agonist inhibited γδ T cell-mediated cytotoxicity. CONCLUSIONS AND IMPLICATIONS: Histamine activated signalling pathways typical of chemotaxis (G(i) protein-dependent actin reorganization, increase of intracellular calcium) and induced migratory responses in γδ T lymphocytes, via the H(4) receptor, whereas it down-regulated γδ T cell mediated cytotoxicity through H(2) receptors and G(s) protein-coupled signalling. Our data suggest that histamine activated γδ T cells could modulate immunological surveillance of tumour tissue.

Taurine trophic modulation of goldfish retinal outgrowth and its interaction with the optic tectum.

Goldfish retinal explant outgrowth in the presence of fetal calf serum is stimulated by taurine. In the absence of it, but with glucose in the medium, length of neurites is still elevated by the amino acid. Using the medium in the presence of glucose, but in the absence of fetal calf serum, we explored the effect of optic tectum medium from cultures of them coming from goldfish without crush of the optic nerve or 3, 5, 10, 14 and 20 days after crush. Retinal explants, intact or from goldfish with crush of the optic nerve 10 days prior to starting the culture, were employed in order to measure the possible effect of optic tectum media and the inter action with taurine. In other type of experiments the optic nerve was crushed 1, 2, 4, 7 and 10 days before dissection of the optic tectum, and then co-cultured with intact or 10 days post-crush retinal explants. Optic tectum media produced a time-dependent effect on outgrowth in lesioned retinas with a maximum effect around 5 days after the lesion for the corresponding optic tectum. Taurine, 4 mM, did not further affect the outgrowth in the presence of optic tectum media, but did significantly increase length of neurites either in intact or in post-lesion retinas. Co-culture of optic tectum at different days post-lesion and retinas at 10 days post-lesion increased the outgrowth around 4 days post-lesion, in a preparation resulting in mutual effects of both types of tissues. The addition of taurine in these conditions did not further increase outgrowth, rather inhibited it according to the time after lesion of optic nerve corresponding to the co-cultured optic tectum. The effect of taurine was concentration-dependent, since 0.2 mM was more effective than 2 or 4 mM in the presence of optic tectum with lesion of 2 days. These results demonstrate the time-course of the regeneration processes in the visual system of goldfish, indicating the crucial periods after crush in which the tectum could produce stimulation and later decrease or no effect on outgrowth from the retina. In addition, they are evidences of the interaction between taurine and optic tectum production of time-produced specific agents. The mechanisms underlying these effects are closely related to calcium, as it was demonstrated by the addition of extracellular or intracellular chelators to the medium, which inhibited the effects of the optic tectum and the trophic properties of taurine in this system. The inhibitor of taurine transport, guanidoethylsulfonate, also decreased the stimulatory effects of the optic tectum and of taurine, indicating an interaction of substances produced by the tectum with taurine, and an effect of taurine mediated through its entrance to the cells. Overall, retinal explants outgrowth in the absence of fetal calf serum, the interaction of agents of the optic tectum and taurine modulates outgrowth from the retina, and these effects are mediated by calcium levels and by the levels of intracellular taurine.

Medium requirements for neuritic outgrowth from goldfish retinal explants and the trophic effect of taurine.

The use of culture media of known composition are necessary for studying the role of trophic molecules. Since most of the in vitro research on regeneration of the optic nerve has been performed in the presence of fetal calf serum, the aim of this study was to obtain a medium in which the neuritic outgrowth from post-crush goldfish retinal explants could take place without adding fetal calf serum. After the lesion of the optic nerve (10 days), the retina of goldfish was dissected and explants were cultured for 5 and 10 days in the absence or in the presence of fetal calf serum, at which time the neuritic outgrowth was determined. Various concentrations and combinations of glucose, albumin, calcium, HEPES and taurine were used. The highest neuritic outgrowth was observed in the presence of fetal calf serum, in which condition the amino acid taurine increased length and density of neurites. Media supplemented with albumin, calcium or HEPES did not modify the outgrowth of neurites from the explants. However, glucose favored the neuritic outgrowth in a bell-shaped manner, although fibers were thinner than those observed in the presence of fetal calf serum. Taurine did not stimulate outgrowth of neurites from explants growing in a medium with optimal concentrations of glucose, indicating that elements of the fetal calf serum are determinant for the trophic effect of taurine. The present results contribute to further studies, such as those related to the effect of taurine and of trophic factors derived from the optic tectum, which would be performed in the presence of a medium free of fetal calf serum.

Sensibilidad a citostáticos de células de tumores cerebrales humanos en cultivo / Sensitivity to cytostatics of cells of human brain neoplasm in culture

Los cultivos de gliomas malignos se han realizado con el objeto de conocer su biología, estudiar nuevas drogas antineoplásicas y determinar la sensibilidad diferencial a citostáticos. Catorce pacientes con diagnósticos clínico y radiológico sugestivos de tumor cerebral maligno fueron seleccionados de acuerdo a criterios específicos, lo que incluyó la evaluación según el esquema de Karnofsky, el cual determina el estado funcional independiente del paciente. Se realizó la resección quirúrgica, se efectuó el estudio histopatológico y se tomó una muestra del tumor para cultivo en monocapas. La confluencia celular se alcanzó entre 15 y 45 días y los cultivos fueron expuestos a los citostáticos (1,3-bis(2-cloroetil)-1- nitrosouréa, cisplatino o vincristina por una hora. Se determinó la integridad de la membrana mediante la exclusión del colorante azúl de Tripán. Se encontraron diferencias estadísticamente significativas entre las drogas utilizadas y entre los tumores, incluso con el mismo diagnóstico histopatológico, lo cual indica la existencia de resistencia diferencial de las células tumorales a los citostáticos utilizados. Este estudio constituye un ensayo que sustenta la realización de evaluaciones a largo plazo con el fin de emplear una quimioterapia específica en pacientes en los que probablemente se combine ésta con la radioterapia

Taurine as a micronutrient in development and regeneration of the central nervous system.

Taurine is an amino acid known to possess trophic properties in the central nervous system. The relevance of its presence in maternal milk is related to its role as an essential nutrient. Taurine deficiency around birth produces anatomical and functional modifications in the brain and in the retina. In addition, taurine favors neuron proliferation and survival, as well as neurite extension. The mechanisms by which taurine exerts its trophic role in the regenerating retina are related to increases in calcium fluxes, to modifications of protein phosphorylation, and to influence of the target organ. Moreover, taurine-zinc interaction might be crucial in the development of structures such as the hippocampal formation. Thus, taurine can be considered as one of the determinant nutritional molecules during development and regeneration of the central nervous system.

Differential taurine effect on outgrowth from goldfish retinal ganglion cells after optic crush or axotomy. Influence of the optic tectum.

The interaction between innervated tissues, targets and nerves is crucial in the maintenance of physiological conditions, and the disturbance of this harmony causes the production of morphological and biochemical changes. After lesion of the optic nerve, several modifications take place in the retina, the optic tectum and the optic nerve. The influence of the tectum on the outgrowth from the goldfish retina and the possible role of taurine was studied. Ganglion retinal cells were identified by retrolabeling with Dil. Crushing the optic nerve 10 days prior to plating retinal cells, as compared with optic axotomy, did not affect the survival of cultured retinal cells, as well as the length of the neurites. However, the number of neurites per cell and the branching of the longest fiber were higher after axotomy than after crushing. The addition of taurine to the medium did not modify this response at 5 days in culture. At early periods in culture, the stimulatory effect on isolated ganglion cell outgrowth produced by taurine was enhanced after axotomy respecting crushing of the optic nerve, but was not affected in retinal explants. The addition of medium from cultured optic tectum several days post-crush of the optic nerve to retinal explants from intact retinas or coming from post-crush retina modified the outgrowth, being inhibitory or stimulatory in a time-dependent manner. The co-culture of optic tectum and retina also affected the outgrowth from the retina with a byphasic shape. The results support the differential response of the retina facing partial or complete interruption with the target and limit the effect of taurine to early periods in culture. In addition, the production of inhibitory factors from the tectum, plus the stimulatory ones, are strongly supported by this work.

Regulation of high affinity taurine transport in goldfish and rat retinal cells.

Adaptive regulation and modulation by phosphorylation are mechanisms by which some cells control taurine transport. Goldfish and rat retinal cells were incubated with the activator of protein kinase C, phorbol 12,13-dibutyrate (PDBu), or the inhibitor of protein phosphatases, okadaic acid (OKA). OKA, 1 nM, inhibited the uptake of taurine at short period of incubation in goldfish retinal cells, and at low concentrations in rat retinal cells incubated with the inhibitor for 1 h. PDBu treatment did not produce significant effects. Isolated Müller cells from the goldfish retina presented a clear adaptive regulation and a decrease of taurine uptake by increasing phosphorylation either by the stimulation of PKC with PDBu or the inhibition of phosphatases with OKA.

Taurine might be acting as a trophic factor in the retina by modulating phosphorylation of cellular proteins.

Protein phosphorylation is involved in the regeneration of the nervous system. Taurine modulates the phosphorylation of specific proteins in the retina, and also increases outgrowth from ganglion cells. In order to test the possible role of protein phosphorylation on the outgrowth from the retina and on the trophic effect of taurine, in vitro studies were performed in the presence of phorbol and nonphorbol protein kinase C activators, the protein kinase C inhibitor tamoxifen, and phosphatase inhibitors. After crush of the optic nerve, explants of the goldfish retina were cultured and the outgrowth was evaluated by measuring the length and the density of neurites. The activation of protein kinase C decreased the outgrowth from the explants and impaired the stimulatory effect of taurine. Phosphatase inhibitors produced a similar effect on basal and taurine-modulated outgrowth. In certain concentrations, some of these drugs did not affect the emission of neurites in the absence of taurine, but decreased the effect of the amino acid. Tamoxifen also reduced the outgrowth, probably acting at other cellular levels or indicating that the regulation of outgrowth by phosphorylation is a complex and dual process. The response to the drugs, evaluated by length or density of fibers, was not the same, since rate of outgrowth was more affected than density, which suggests that both parameters are modulated at differential stages or sensitivities to the tested agents.

Taurine-stimulated outgrowth from the retina is impaired by protein kinase C activators and phosphatase inhibitors.

Taurine increases neurite elongation of post-crush goldfish retinal explants, as well as the number of outgrowing isolated cells from goldfish and rat retina in culture. The trophic effect of taurine is related to an elevation in calcium flux rather than increased cell proliferation. Since taurine regulates phosphorylation in rat retina, we investigated if this process could be involved in the mechanism of taurine action on outgrowth. Control and taurine-supplemented post-crush goldfish retinal explants were cultured in the presence of protein kinase C activators or phosphatase inhibitors, and the length of neurites was measured after five days in culture. In some cases, there was an inhibition of the stimulatory effect of taurine without a modification in basal outgrowth. In others, outgrowth of control explants was also reduced. A certain level of protein phosphorylation seems to be critical for the trophic effect of taurine in the retina.

Differential effect of taurine and serotonin on the outgrowth from explants or isolated cells of the retina.

The sulfur amino acid taurine and the indoleamine serotonin increases and decreases, respectively, the outgrowth from goldfish retinal explants. Taurine seems to be acting, at least partially, through an increase in calcium fluxes, and the serotonin-inhibiting effect appears to be mediated by serotonin1A receptors and cAMP. Isolated cells of postcrush goldfish retina and of retina from 5-day-old rats were cultured in the presence of taurine or serotonin. In the goldfish, the classical morphology of postcrush ganglion cells was observed. An antibody against the glycoprotein Thy-1 labelled three types of cells in the cultures of goldfish retina. The number of cells outgrowing and the length of the main neurite was measured at 5 days in culture in both species. The number of cells presenting neurites was increased in the goldfish retina by the addition of taurine, and decreased by serotonin. However, the length of the neurites was unaffected by the addition of the modulators. In the rat, only a slight decrease in the number of cells outgrowing was observed in the presence of serotonin. The incorporation of [3H]thymidine was not modified after 5 days in culture in the presence of taurine or serotonin, either in the goldfish or in the rat retina. The antibody Thy 1.1 can label retinal cells of the goldfish in vitro, one of them being ganglion cells. The trophic effect exerted by taurine in the postcrush goldfish retina needs the integrity of the tissue favoring the interaction of cells and factors, because outgrowth increases in retinal explants, but not in isolated cells.

Effect of blocking TNF on IL-6 levels and metastasis in a B16-BL6 melanoma/mouse model.

We studied the relationship between tumour necrosis factor (TNF) and interleukin 6 (IL-6) levels, and the metastatic process in C57BL/6 mice after intravenous inoculation of B16-BL6 melanoma cells. Bioactive TNF was not detectable in the sera of inoculated mice, but these animals did show higher TNF levels following intraperitoneal challenge with lipopolysaccharide (LPS) compared to control animals. Serum IL-6 levels were increased in inoculated animals. Injection of a hybrid molecule (p55-sf2) composed of the human p55 TNF receptor extracellular domain coupled to a human constant region backbone, decreased serum TNF (after LPS challenge) and IL-6 levels in inoculated animals. Lung metastases at 7-14 days were reduced, compared to human IgG-injected control animals, but this effect was lost at day 21 postinoculation. The results suggest that the reduction in the number of metastases may be related to the effect of blocking TNF activity.