El Diario de Pontevedra : periódico liberal: Ano LI Número 15301 - 1937 novembro 2

Fig. S1. Viability of FaDu and Cal27 cells at low concentrations of methylglyoxal. Assessment of the cytotoxic effect of MG at low concentration in a colony-forming assay. (TIFF 92 kb

Perfiles paleokarsticos en el techo de la unidad intermedia del mioceno de la cuenca de Madrid

An intra-Vallesian (Upper Miocene) paleokarst developed at the top of the Intermediate Miocene Unit in the continental Madrid basin is recognized. This paleokarst is a early shallow, tabular-shaped karst that shows a marked control by the depositional facies pattern and lithologies. By integrating morphological, petrological and geochemica1 data, three hydrogeological or environmental zones have been established throughout the paleokarstic profiles: (i) a vadose zone, characterized by vertically elongated caves and discontinuous speleothems and vadose cements (ii) a 3-7 m thick water table fringe, characterized by the wide development of stratiform breccia bodies, the superimposition of both vadose and phreatic features, and the lowest Fe and Mn contents in host-rock carbonates; and (iii) a phreatic zone characterized by an increase of 6I3C values and the predominance of phreatic cementation. The paleogeographic reconstruction for the intra-Vallesian paleokarst using paleokarstic profiles reveals relative topographic highs to the north and topographic lows to the south drawing the paleokarst landscape.<br><br>En el techo de la Unidad Intermedia del Mioceno de la Cuenca de Madrid se ha desarrollado un paleokarst temprano, somero y de forma tabular que muestra un marcado control litol&#243;gico y del dispositivo de facies deposicionales en su g&#233;nesis. Integrando criterios geomorfol&#243;gicos, petrogr&#225;ficos y geoqu&#237;micos se ha establecido una zonaci&#243;n hidrogeol&#243;gica en los perfiles paleok&#225;rsticos estudiados, diferenci&#225;ndose: (i) una zona vadosa caracterizada por la existencia de cavidades alargadas en la vertical tapizadas discontinuamente por espeleotemas y otros cementos vadosos; (ii) una franja de oscilaci&#243;n del nivel fre&#225;tico de unos 3-7 metros de espesor, caracterizado por el desarrollo extensivo de cuerpos brechoides estratiformes, la yuxtaposici&#243;n de cementos vadosos y fre&#225;ticos y los contenidos m&#225;s bajos en Fe y Mn en el material encajante, y (iii) una zona fre&#225;tica caracterizada por un aumento en los valores de 613C y el predominio de la cementaci&#243;n fre&#225;tica. La correlaci&#243;n de los perfiles paleok&#225;rticos revela una paleogeograf&#237;a para el techo de la Unidad Intermedia con un paisaje topogr&#225;ficamente descendente de norte a sur en la cuenca para el Vallesiense

Additional file 7: of Glyoxalase 1 expression is associated with an unfavorable prognosis of oropharyngeal squamous cell carcinoma

Table S5. Univariate analysis of distinct risk factors for progression-free and disease-specific survival. (DOCX 72 kb

Methylglyoxal is an intracellular TRPA1 agonist.

<p>(A) Application of MG evokes concentration-dependent [Ca²⁺]_i-responses in CHO cells expressing mouse or human TRPA1. (B) MG (0.5 mM) elicits inward currents with the characteristic rapid onset and inactivation in the presence of 2 mM Ca²⁺ (top panel). In Ca²⁺-free solutions (1 mM EGTA) the inward current grew more slowly and addition of Ca²⁺ produced the typical current surge followed by a rapid inactivation of the current (bottom panel). (C) MG rapidly activates TRPA1 in excised inside-out patches, with a markedly higher potency than when applied extracellularly (D, compare with A). (E) The current voltage relationship in membrane patches containing several channels demonstrated a reversal potential close to 0 mV and channel block or inactivation at positive membrane potential. (F) Current-voltage relationship for a single channel in an inside-out patch stimulated with MG.</p

Methylglyoxal produce persistent sensory neuropathy.

<p>(A) Intraplantar injections of MG (250 nmoles in 25 µl) evokes nociceptive behaviors in wildtype mice but is without effect in <i>Trpa1^−/−</i> mice (n = 6). Injections of the GLO-1 inhibitor Sr-p-Bromobenzylglutathione cyclopentyl diester (GLOI, 50 mg/kg, 1h before test) did not acutely affect the withdrawal threshold in the paw pressure (B) or von Frey (C) tests compared to mice treated with vehicle (n = 6). (D-G) Effect of GLOI or vehicle injections administered every 2 days during a 2 week treatment period on the paw withdrawal latency from a 50°C hotplate (D), 10°C coldplate (E), the paw withdrawal threshold in the paw pressure test (F) and in response to stimulation with von Frey filaments (G). The data points are mean ± SEM from n = 6 mice, except for the last time point (15 days after the final injection) where n = 3. Data were analyzed by t-test (panel A) or ANOVA followed by Tukey’s HSD test (**P<0.01, ***P<0.001 compared to vehicle treated group).</p

Methylglyoxal stimulates DRG neurons expressing TRPA1.

<p>(A) Pseudocoloured images illustrating [Ca²⁺]_i-concentrations in Fura-2 loaded cultured DRG neurons. MG evokes [Ca²⁺]_i-responses in TRPA1 containing neurons. (B) Typical examples of [Ca²⁺]_i-responses in capsaicin sensitive DRG neurons from <i>Trpa1^+/+</i> and <i>Trpa1^−/−</i> mice. (C) MG activates characteristic TRPA1 currents in DRG neurons (holding potential -60 mV). Before addition of 2 mM Ca²⁺ this neuron was superfused with a solution containing 15 µM Ca²⁺, which prevents the Ca²⁺-induced current surge.</p

Intra-epidermal nerve fiber density is unaffected by 2 weeks GLO-1 inhibition.

<p>(A) PGP9.5 positive nerve fibers crossing the basal membrane into the epidermis (the white arrow highlights an example) in 8 µm sections of plantar skin (scale bar = 20 µm). (B) Following two weeks treatment with the GLO-1 inhibitor (GLO-I, 50 mg/kg every other day), the number of intra-epidermal nerve fibers crossing into the epidermis/mm skin was unchanged in both <i>Trpa1^+/+</i> mice and in <i>Trpa1^−/−</i> littermates. Data were analyzed by ANOVA followed by Tukey’s HSD test (n = 6 mice, *P<0.05 compared to vehicle treated <i>Trpa1^+/+</i> mice).</p

TRPA1 is expressed in MDCK cells.

<p>(A) The TRPA1 agonists AITC, H₂O₂, MG and cinnamaldehyde stimulate [Ca²⁺]_i-responses in MDCK cells. (B, C) The selective TRPA1 antagonist AP18 (10 µM) produces a rightward shift of AITC and MG evoked [Ca²⁺]_i-responses in MDCK cells.</p

Methylglyoxal is a reversible TRPA1 agonist.

<p>The cysteine substitutions C665S, C641S and C621S (in human TRPA1) do not significantly affect responses evoked by MG (A), but dramatically reduce the sensitivity to AITC (B). (C) The lysine substitution K712Q (mouse TRPA1) reduced [Ca²⁺]_i-responses evoked by AITC (50 µM). (D) The K712Q mutation reduced the amplitude of [Ca²⁺]_i-responses evoked by MG (2 mM) and Δ⁹-tetrahydrocannabiorcol (20 µM; C16) in individual CHO cells (n = 22–64). (E) Outward TRPA1 current responses (+60 mV) evoked by MG (1 mM) in CHO cells decay relatively slowly when MG is removed (recorded under Ca²⁺-free conditions). Application of DTT (2 mM) produces a rapid, partial current inactivation. (F) Currents evoked by AITC (50 µM) under the same conditions inactivate relatively slowly, but are resistant to DTT and leave TRPA1 refractory to stimulation. (G) Currents elicited by the oxidant H₂O₂ (5 mM) remain at a stable level after removal of H₂O₂, but are rapidly and reversibly inactivated by DTT. Data were analyzed by ANOVA followed by Dunnett’s <i>post-hoc</i> test (panel B) or by t-test (*P<0.05, **P<0.01, ***P<0.001 compared to the wildtype channel).</p

TRPA1 does not influence insulin release.

<p>(A) MG (0.5 mM) evokes indistinguishable [Ca²⁺]_i-responses in β-cells from <i>Trpa1^+/+</i> (black) and <i>Trpa1^−/−</i> (red) mice (n = 15-28). (B) Illustration of the time course of MG and tolbutamide (200 µM) evoked [Ca²⁺]_i-responses in β-cells. (C, D) Effect of the TRPA1 antagonist HC030031 (HC, 100 mg/kg) or vehicle (Veh) administered 30 min before glucose or vehicle in the glucose tolerance test (2 g/kg, <i>i.p.</i>) performed in <i>Trpa1^+/+</i> (C) and <i>Trpa1^−/−</i> mice (D). Data points are mean ± s.e.m of n = 6 mice and were analyzed by ANOVA followed by Tukey’s HSD test (*P<0.05, **P<0.01, ***P<0.001 compared to the group treated with vehicle and glucose, †P<0.05 compared to the group treated with vehicle and vehicle).</p