Tissue damage markers after a spinal manipulation in healthy subjects: a preliminary report of a randomized controlled trial.

Spinal manipulation (SM) is a manual therapy technique frequently applied to treat musculoskeletal disorders because of its analgesic effects. It is defined by a manual procedure involving a directed impulse to move a joint past its physiologic range of movement (ROM). In this sense, to exceed the physiologic ROM of a joint could trigger tissue damage, which might represent an adverse effect associated with spinal manipulation. The present work tries to explore the presence of tissue damage associated with SM through the damage markers analysis. Thirty healthy subjects recruited at the University of Jaén were submitted to a placebo SM (control group; n = 10), a single lower cervical manipulation (cervical group; n = 10), and a thoracic manipulation (n = 10). Before the intervention, blood samples were extracted and centrifuged to obtain plasma and serum. The procedure was repeated right after the intervention and two hours after the intervention. Tissue damage markers creatine phosphokinase (CPK), lactate dehydrogenase (LDH), C-reactive protein (CRP), troponin-I, myoglobin, neuron-specific enolase (NSE), and aldolase were determined in samples. Statistical analysis was performed through a 3 × 3 mixed-model ANOVA. Neither cervical manipulation nor thoracic manipulation did produce significant changes in the CPK, LDH, CRP, troponin-I, myoglobin, NSE, or aldolase blood levels. Our data suggest that the mechanical strain produced by SM seems to be innocuous to the joints and surrounding tissues in healthy subjects.

Age and sex-related serum changes in nitric oxide: correlations with serological markers.

Serum nitric oxide levels, systematically determined in 200 men and women from 18 to 65 year-old, undergo age and sex changes that strongly correlate with serological markers such as those related with cardiovascular functions and lipid profile.

Valoración de la determinación de la enzima lactato deshidrogenasa en los trastornos hipertensivos del embarazo / Evaluation of the determination of the lactate-dehydrogenase enzyme in the hypertensive diseases of pregnancy

Introducción: Los trastornos hipertensivos del embarazo (THE), en general, y la preeclampsia, en particular, son algunas de las complicaciones más graves que pueden afectar a madre y feto. Son muchas las pruebas clínicas y de laboratorio que se han utilizado en busca de un indicador de afección de los THE. Entre los segundos destaca la enzima lactato deshidrogenasa (LDH). Objetivo: Valorar la correlación de las distintas formas isoenzimáticas de LDH con los distintos parámetros indicativos de afección y gravedad en los casos de THE. Material y método: Se han estudiado la enzima LDH y cada una de sus 5 isoformas, en 30 pacientes, a las que, en el curso de su gestación, se les encontraron valores de presión arterial (PA) máxima de 140 mmHg y mínima de 90 mmHg. Las determinaciones se efectuaron en suero sanguíneo y la extracción se realizó antes de las 24 h de su ingreso. Resultados: No encontramos correlación entre la LDH total ni de las isoformas 1, 3 y 4, con ninguno de los parámetros estudiados. La LDH-2 sí mostró una fuerte correlación con la transaminasa glutámico-oxalacética (GOT) y la transaminasa glutámico-pirúvica (GPT). La LDH-5 mostró correlación con la proteinuria y la GOT, así como con la PA máxima y el hematocrito. No encontramos correlación alguna entre las PA máxima y mínima (r = 0,266; p = 0,156; n = 30), ni de la máxima con ninguno de los parámetros bioquímicos o hematológicos evaluados. En cambio, encontramos una fuerte correlación positiva entre la PA mínima y los valores de ácido úrico y GOT, que casi fue significativa para la GPT (p = 0,06). Conclusiones: Consideramos que la determinación de la LDH-5 en los casos de THE y su aumento en suero son indicadores, al presentar una fuerte correlación con alteraciones importantes en este cuadro, como la proteinuria, y con las alteraciones hepáticas y hematológicas descritas en esta enfermedad (AU)

Introduction: Hypertensive disorders of pregnancy (HDP) and pre-eclampsia in particular, are some of the most serious complications that can affect mother and foetus. Numerous clinical and laboratory tests have been used in the search for an indicator for HDP; amongst the latter, lactate-dehydrogenase enzyme stands out. Objective: To evaluate the correlation of the different isoenzymatic forms of LDH (lactate-dehydrogenase) with the different indicative parameters of affectation and severity for the cases with HDP. Material and methods: Lactate-dehydrogenase enzyme and each of its 5 isoenzymes was studied in 30 patients who, during the course of their pregnancies, were found to have arterial pressure with a maximum value of 140 mm Hg and a minimum value of 90 mm Hg. Measurements were made using serum extracted within 24 hours of their admission. Results: We did not find any correlation between the total LDH and the isoenzymes 1,2,3 and 4 in any of the parameters studied. LDH-2 showed a strong correlation with GOT and GPT. LDH-5 showed correlation with proteinuria and GOT and also with maximum arterial tension and haematocrit. We did not find any correlation between maximum and minimum arterial tension (r=.266, p=.156, n=30), nor of the maximum arterial tension with any of the biochemical or haematological parameters evaluated. However we found a strong positive correlation between minimum arterial tension and uric acid and GOT values and that was almost significant for GPT (p=.06). Conclusions: We consider that the determination of LDH-5 in cases of HDP, and its increase in serum, is an indicator as it presents a strong correlation with important alterations in this clinical picture, such as the proteinuria and the hepatic and haematological alterations described in this pathology (AU)

Electroforesis en acetato de celulosa de sueros congelados / Electrophoresis on cellulose acetate of frozen serums

Se estudió el efecto de la congelación prolongada de sueros humanos sobre la electroforesis de proteínas. Para ello se conservaron 125 sueros a -30 °C durante 15 meses, realizándose un fraccionamiento antes y después de la congelación. La comparación entre los valores medios obtenidos en ambos momentos mostró diferencias estadísticamente significativas en cada uno de los parámetros electroforéticos estudiados. Se apreció una tendencia al incremento en la región de las globulinas y un descenso de la banda de albúmina. Estas variaciones superaron el 5 por ciento de los valores basales en más de la mitad de las muestras. Sin embargo, sólo originaron cambios clínicamente relevantes (reconsideración :como patológicos de resultados previamente normales) en la banda de y-globulinas (AU)

Efectos de la congelación sobre las proteínas totales / Effects of the freezing on total proteins

La congelación es un método muy utilizado para conservar las muestras biológicas y evitar los fenómenos de degradación. Sin embargo, se han descrito alteraciones tras la realización de determinaciones en muestras congeladas. Estudiamos los niveles de proteínas totales en 125 sueros congelados I S meses a -30 °C. Se encontró un aumento significativo de la concentración plasmática respecto a valores basales en el global de muestras analizadas y en el subgrupo de muestras normoproteicas. En el test de regresión lineal se obtuvo un coeficiente R2 de 0,8736. Se observaron diferencias superiores al 5 por ciento de los valores basales en más de la mitad de las muestras, aunque ello no influyó significativamente en su clasificación como normales o patológicas. La congelación prolongada a -30 °C no siempre evita la presencia de interferencias, hecho que hay que considerar al interpretar los resultados obtenidos con muestras conservadas (AU)

S-adenosylmethionine prevents hepatic tocopherol depletion in carbon tetrachloride-injured rats.

In various experimental models, S-adenosylmethionine (SAMe) has been shown to reduce liver injury by preventing depletion of glutathione, one of the antioxidant systems that plays a critical role in defence against oxidative stress. On the other hand, alpha-tocopherol may be decreased in liver diseases, and treatment with this vitamin reduces liver injury in CCl(4)-treated rats. Since there is a close relationship among the different antioxidant systems (mainly glutathione, alpha-tocopherol and ascorbic acid), we have assessed whether, as well as restoring hepatic glutathione content, SAMe has any effect on liver alpha-tocopherol and ascorbic acid levels in CCl(4)-injured rats. Four groups of seven male Wistar rats treated for 9 weeks were studied: rats induced to cirrhosis with CCl(4), rats induced to cirrhosis plus SAMe administration (10 mg x kg(-1) x day(-1)) and their respective controls. Liver samples were obtained for measuring levels of glutathione, alpha-tocopherol, ascorbic acid and thiobarbituric acid-reactive substances (TBARS), and hydroxyproline concentration as an index of collagen content. The hydroxyproline content was higher in CCl(4)-injured rats than in the control group (4.4+/-1.8 and 1.1+/-0.3 micromol/g respectively; P<0.05). In CCl(4)-injured rats, SAMe administration decreased collagen content (2.7+/-1.0 microl/g; P<0.05) and TBARS, and corrected glutathione depletion. alpha-Tocopherol was significantly lower in CCl(4)-injured rats than in controls (17.3+/-4.9 and 23.0+/-4.0 micromol/g respectively; P<0.05). By contrast, alpha-tocopherol levels were similar (23.8+/-5.1 micromol/g) in CCl(4)-injured rats receiving SAMe and in controls. In CCl(4)-injured rats, liver ascorbic acid was decreased in comparison with controls (4.9+/-1.8 and 8.2+/-1.0 micromol/g respectively; P<0.05), levels which were not replenished by SAMe (4.6+/-0.4 micromol/g). In conclusion, SAMe not only decreases fibrosis and protects against hepatic glutathione depletion, but has a further antioxidant effect of preventing alpha-tocopherol depletion in CCl(4)-injured rats.

Enhanced DNA binding and activation of transcription factors NF-kappa B and AP-1 by acetaldehyde in HEPG2 cells.

Because transcription factors NF-kappaB and activator protein-1 (AP-1) are known to regulate gene expression, we have analyzed the role of acetaldehyde in the activation of NF-kappaB and AP-1 in HepG2 cells. Binding activity and transactivation of NF-kappaB and AP-1 were determined by gel retardation assays and transfection of a luciferase reporter construct controlled by kappaB and AP-1 binding sites, respectively. Acetaldehyde enhanced the DNA binding of NF-kappaB and AP-1 by 1 and 4 h, respectively, increasing the kappaB- and AP-1-dependent luciferase expression. Supershift assays revealed the presence of NF-kappaB heterodimers p65/p50 and p50/p52, whereas nuclear c-Jun levels correlated with the DNA binding of AP-1. The enhanced binding of NF-kappaB to DNA by acetaldehyde in intact cells was accompanied by the proteolytic degradation of IkappaB-alpha. However, the addition of acetaldehyde to cytostolic extracts from untreated Hep G2 cells did not affect the DNA binding of AP-1 but activated the NF-kappaB heterodimer p65/p50 in the absence of IkappaB-alpha degradation. Preincubation of HepG2 cells with protein kinase C inhibitors abolished the enhanced DNA binding of NF-kappaB and AP-1 caused by acetaldehyde. Hence, these findings uncover a previously unrecognized role for acetaldehyde in the activation of NF-kappaB and AP-1, which may be of relevance in the alcohol-induced liver disease.

Influencia de la congelación a corto y largo plazo sobre la concentración de proteínas totales y su electroforesis / Influence of freezinf to short and long term on the concentration and electrophoresis of total proteins

La congelación es el método más utilizado para la conservación de las muestras biológicas aunque no siempre asegura la estabilidad de las mismas. Se estudiaron, mediante el método de Biuret y un fraccionamiento electroforético, 40 sueros congelados a 30 ºC durante 2 y 15 meses. Al comparar los valores obtenidos tras el procesamiento inmediato con los encontrados tras las congelaciones no se observaron diferencias en la concentración de proteínas totales pero sí cambios en el valor medio de cada parámetro electroforético. Estas modificaciones fueron especialmente evidentes en las bandas de alfa-1, alfa-2 y gammaglobulinas, con incrementos que oscilaron entre el 6,7 y 18,9 por ciento de los valores previos. A pesar de ello, estas variaciones no indujeron cambios significativos en la consideración de las muestras como "normales" o "patológicas" antes y después de las congelaciones (AU)

Carbohydrate-deficient transferrin as a marker of alcohol consumption in male patients with liver disease.

Carbohydrate-deficient transferrin (CDT) has been proposed as a marker of alcohol abuse. However, its value in patients with associated liver disease is still controversial. The aim of the study was to investigate the usefulness of CDT as a marker of alcohol consumption in patients with liver disease. We measured serum levels of CDT and those of commonly used hematological and biochemical markers, mean corpuscular volume (MCV), transaminases (AST and ALT), and gamma-glutamyltransferase in 179 male subjects divided into four groups: 45 active drinkers (13 with normal liver, 21 with fibrosteatosis, and 11 with liver cirrhosis), 45 abstinent chronic alcoholics (18 with and 27 without liver disease), 58 patients with nonalcoholic liver disease, and 31 healthy controls. Serum CDT in active alcoholics was 37.5 +/- 3.6 units/liter, being significantly higher than that of abstinent alcoholics (20.3 +/- 1.5 units/liter), patients with nonalcoholic liver disease (18.1 +/- 1.1 units/liter), and controls (13.1 +/- 0.8 units/liter). Contrary to the other markers, no significant differences were observed in CDT values in relation with the presence and severity of liver disease in either the active drinkers or in the abstinent alcoholics. The sensitivity and specificity of CDT as a marker of alcoholism in the series as a whole was 64% and 82%, respectively, similar to the best conventional marker, MCV (64 and 82%). In patients with liver disease, CDT maintained good sensitivity (72%) and specificity (83%). Receiver operating characteristic analysis confirmed that CDT had a similar diagnostic value to that of MCV, but better than gamma-glutamyl-transferase and transaminases for the detection of alcohol abusers. The good diagnostic efficacy of CDT remained unchanged when analyzing only patients with liver disease. We conclude that serum CDT is a good marker of alcoholism and is less influenced than the currently used biochemical markers for associated liver disease. Thus, CDT is an effective laboratory test to detect alcohol abuse regardless of the presence of alcoholic liver disease.

Effects of S-adenosylmethionine on lipid peroxidation and liver fibrogenesis in carbon tetrachloride-induced cirrhosis.

BACKGROUND/AIM: The aim of this study was to investigate the effects of S-adenosylmethionine on liver peroxidation and liver fibrogenesis in carbon tetrachloride-induced cirrhosis. METHODS: Cirrhosis was induced in three groups of six rats by repeated injections of carbon tetrachloride over 9 weeks. One group of animals was treated only with carbon tetrachloride, and the other two received carbon tetrachloride plus S-adenosylmethionine (10 mg/kg intramuscularly daily) from week 3 to week 9, and from week 6 to week 9 of the study, respectively. Two additional groups of six rats, a control group and a group treated only with S-adenosylmethionine, were also studied. Glutathione concentration, thiobarbituric acid-reactive substances, collagen content, prolyl hydroxylase activity, and procollagen type I mRNA expression were determined in liver samples. RESULTS: All carbon tetrachloride-treated rats had cirrhosis at the end of the study. Cirrhosis was also present in five of the six carbon tetrachloride-treated rats receiving S-adenosylmethionine for 3 weeks, but in only one of the six rats that received S-adenosylmethionine for 6 weeks. Hepatic glutathione was significantly diminished in carbon tetrachloride-treated rats (2.7 +/- 0.3 mumol/g tissue) and returned to normal in rats receiving S-adenosylmethionine for 3 or 6 weeks (3.7 +/- 0.13 and 3.9 +/- 0.11 mumol/g tissue, respectively). The hepatic thiobarbituric acid-reactive substances were significantly lower in rats treated with carbon tetrachloride and S-adenosylmethionine for 6 weeks (98 +/- 5 nmol/g) than in rats treated with carbon tetrachloride (134 +/- 12 nmol/g) and in those treated with carbon tetrachloride and S-adenosylmethionine for 3 weeks (127 +/- 13 nmol/g). There were no differences in either hepatic collagen and prolyl hydroxylase activity between rats that received only carbon tetrachloride and those treated with S-adenosylmethionine for 3 weeks. In contrast, carbon tetrachloride-treated rats receiving S-adenosylmethionine for 6 weeks had significantly lower collagen and prolyl hydroxylase activity than the other two groups. A much greater increase in procollagen type I mRNA was found in carbon tetrachloride-treated rats than in rats treated with carbon tetrachloride and S-adenosylmethionine for 6 weeks. Furthermore, there was a significant correlation between the hepatic thiobarbituric acid-reactive substances and prolyl hydroxylase activity and hepatic collagen. CONCLUSIONS: We conclude that the early administration of S-adenosylmethionine in a model of carbon tetrachloride-induced liver injury restores glutathione levels and reduces lipid peroxidation, resulting in less advanced liver fibrosis.

Carbon tetrachloride-induced hepatic injury is associated with global DNA hypomethylation and homocysteinemia: effect of S-adenosylmethionine treatment.

Carbon tetrachloride (CCl4) administration to rats produces hepatic cirrhosis and supplementation with S-adenosylmethionine (SAM) can partially prevent CCl4-induced liver injury. These effects are thought to be caused by oxidative stress and the subsequent formation of free radicals, but the mechanism whereby this occurs and the accurate nature of the mechanisms by which SAM exerts its protective action are not well understood. The effect of short-term administration of CCl4 on hepatic DNA methylation and on SAM and S-adenosylhomocysteine (SAH) were assessed. CCl4 administration to rats for 3 weeks resulted in hypomethylation of liver DNA, determined by comparing the extent to which DNA from livers of control or treated animals could be methylated in vitro using [3H-methyl] SAM as methyl donor. This CCl4 effect on DNA methylation was corrected by the administration of SAM (10 mg/kg/d, intramuscularly), with values of methyl groups incorporation comparable with those observed in the control animals. hepatic SAM was decreased by CCl4 (65.3 +/- 5.27 vs. 102.2 +/- 4.89 nmol/g; P < .05) and SAH was increased (69.5 +/- 14.6 vs. 29.4 +/- 3.83 nmol/g; P < .05). This led to a marked reduction of the SAM/SAH ratio (the methylation ratio) from 3.47 in control rats to 0.94 in CCl4-treated animals (P < .05). SAM treatment partially prevented (P < .05) the reduction of the ratio SAM/SAH induced by CCl4. CCl4 also induced a marked elevation of serum homocysteine levels (more than 20-fold; P < .001), which was partially prevented by SAM administration.(ABSTRACT TRUNCATED AT 250 WORDS)

[Predisposing factors for the development of acute severe tubular necrosis in the immediate post-transplantation period]. / Factores predisponentes al desarrollo de necrosis tubular aguda severa en el postrasplante inmediato.

Retrospective study on the incidence of severe acute tubular necrosis (ATN), defined as the need for dialysis on the first week post-transplantation ruling out acute rejection or technical complication, in our series of 81 renal transplantations from corpse donor. It includes an evaluation of the influence on severe ATN presentation of parameters such as recipient and donor's age, level of plasma creatinine prior to extraction, whether the corpse had hypotension, duration of both vascular anastomosis and procedure, type of removal (single or multiorgan) and cold ischaemia. We conclude that in our experience, cold ischaemia of the graft is the only determining factor among the cases studied for ATN presentation conditioning a reduced long-term survival of the graft (0.758 vs 0.971 at 18 months: p.05). Peripheral blood typing of corpse donor would allow us to shorten cold ischaemia duration, and thus achieve a low rate of severe ATN (13.6%).