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Nansi López-Valverde,1 Antonio López-Valverde,1 Rafael Gómez de Diego,2 Clara Cieza-Borrella,3 Juan M Ramírez,4 Rogelio González-Sarmiento31Dental Clinic, Department of Surgery, Biomedical Research Institute of Salamanca (IBSAL), Salamanca, Spain; 2Dental Clinic, Alfonso X El Sabio University, Madrid, Spain; 3Molecular Medicine Unit, Department of Medicine, Biomedical Research Institute of Salamanca (IBSAL), Salamanca, Spain; 4Department of Morphological Sciences, School of Medicine, University of Córdoba, Córdoba, SpainAims: In this study we wanted to figure out if there was a correlation between OPRM1 N40D, TRPV1 I316M, TRPV1 I585V, NOS3 −786T>C and IL6 −174C>G polymorphisms and the response to locally applied articaine-epinephrine anesthetic.Methods: In this observational study, 114 oral cell samples of patients anesthetized with articaine-epinephrine (54 from men 60 from women), were collected from dental centers in Madrid (Spain). High molecular weight DNA was obtained from oral mucosa cells. The analysis of OPRM1 N40D (rs1799971), TRPV1 I316M (rs222747), TRPV1 I585V (rs8065080) and IL6 −174C>G polymorphism was performed through real-time PCR allelic discrimination using TaqMan probes. Polymorphism NOS3 −786T> C (rs2070744) was analyzed using RFLP-PCR.Results: The studied polymorphisms are involved neither in the response to the anesthetic, nor in the intensity of perceived dental pain. However, in a subset of female patients we found that TRPV1 I316M was associated with a delayed onset of anesthesia.Conclusions: There is no association among these polymorphisms and the time elapsed between the application of the anesthetic and the onset of its effect.Keywords: Pain, polymorphism, OPRM1, TRPV1, NOS3, IL6

Salvador Pastor-Idoate,1,2 Irene Rodríguez-Hernández,2,3 Jimena Rojas,1 Lucia Gonzalez-Buendia,1 Santiago Delgado-Tirado,1,4 Jose Carlos López,1 Rogelio González-Sarmiento,2,3 Jose C Pastor1,4 1IOBA Eye Institute, University of Valladolid, Valladolid, 2Molecular Medicine Unit, Department of Medicine, 3Molecular and Cellular Cancer Biology Institute, High Council of Scientific Research, Biomedical Research Institute of Salamanca, University of Salamanca, Salamanca, 4Department of Ophthalmology, Hospital Clínico Universitario, Valladolid, Spain Purpose: The objective of this study is to determine the expression and localization of lymphotoxin alpha (LTA) in human retinas and the functionality of one of its polymorphisms rs2229094 (C13R) (T>C), previously associated with proliferative vitreoretinopathy (PVR) development.Materials and methods: Total RNA from three healthy human retinas were extracted and subjected to reverse transcription-polymerase chain reaction (RT-PCR) analysis, using flanking primers of LTA cDNA. In addition, three human eyes with retinal detachment (RD) and three healthy control eyes were subjected to immunohistochemistry (IHC) with a specific antibody against LTA. The functionality of T and C alleles was assessed by using pCEFL-Flag expression vector and transient transfection assays in COS-1 cell line. In addition, expression analysis by RT-PCR, Western blot and subcellular localization of both alleles and by immunofluorescence assay was performed.Results: RT-PCR analysis revealed no significant levels of messenger RNA (mRNA) LTA in healthy human retinas. Sequential IHC staining showed differences between healthy human and RD retinas. No differences in mRNA and protein expression levels and in subcellular localization between both alleles were found. Both alleles were located in the cytoplasm of COS-1 cells.Conclusion: Although results suggest lack of functionality, the differences found in IHC study and its strong association with PVR and its relationship with tumor necrosis factor locus, warrant further studies and could justify the use of this polymorphism as a valid biomarker to identify high-risk patients to develop PVR after RD. Keywords: proliferative vitreoretinopathy, lymphotoxin alpha, tumor necrosis factor alpha, inflammation, cytokines, polymorphism

Fundamento: La enfermedad ósea de Paget (EOP) es un trastorno focal del hueso con aumento el número, tamaño y actividad de los osteoclastos. Algunos datos epidemiológicos apoyan la teoría de su relación con agentes ambientales tóxicos o infecciosos. Su interacción con algunas alteraciones genéticas predisponentes conducirían a la EOP. Las glutatión-S-transferasas (GST) intervienen en la metabolización de toxinas, al catalizar el ataque nucleofílico del sustrato fisiológico, glutatión reducido o GSH (g-Glu-Cys-Gly) sobre el centro electrófilo de un gran número de estructuras tóxicas. Estudiamos si la variabilidad de los genes GSTM1, GSTP1 y GSTT1 se relaciona con el riesgo a desarrollar EOP.Pacientes y métodos: Analizamos a 148 pacientes diagnosticados de EOP y a 207 individuos controles pareados en sexo y edad sin antecedentes de alteraciones óseas. Con DNA genómico obtenido de sangre periférica se estudió la presencia-ausencia de deleción en los genes GSTM1 y GSTT1, mediante PCR multiplex. El estudio del polimorfismo Ile105Val del gen GSTP1 se llevó a cabo mediante PCR y posterior digestión con la enzima de restricción BsmaI. Se analizó la distribución de genotipos mediante el test chi-cuadrado de Pearson. Cuando se encontraron diferencias estadísticamente significativas, realizamos una regresión logística multivariante para conocer el riesgo que puede generar la presencia de un determinado genotipo. Utilizamos el programa SPSS 21.0. Se consideraron diferencias estadísticamente significativas aquéllas con valores de p