Interaction of L-phenylalanine with carbonyl groups in mixed lipid membranes.

The interaction of L-Phe with the membrane components, i.e., lipids and proteins, has been discussed in the current literature due to the interest to understand the effect of single amino acids in relation to the formation of amyloid aggregates. In the present work, it is shown that L-Phe interacts with 9:1 DMPC (1,2-dimyristoyl-sn-glycero-3 phosphocholine)/DPPC (1,2-dipalmitoyl-sn-glycero-3 phosphocholine) mixtures but not in the 1:9 one. An important observation is that the interaction disappears when DPPC is replaced by diether PC (2-di-O-hexadecyl-sn-glycero-3-phosphocholine) a lipid lacking carbonyl groups (CO). This denotes that CO groups may interact specifically with L-Phe in accordance with the appearance of a new peak observed by Infrared spectroscopy (FTIR-ATR). The interaction of L-Phe affects the compressibility pattern of the 9:1 DMPC/DPPC mixture which is congruent with the changes observed by Raman spectra. The specific interaction of L-Phe with CO, propagates to phosphate and choline groups in this particular mixture as analyzed by FTIR-ATR spectroscopy and is absent when DMPC is dopped with diether PC.

Interfacial Hyperactivation of Candida rugosa Lipase onto Ca2Fe2O5 Nanoparticles: pH and Ionic Strength Fine-Tuning to Modulate Protein-Support Interactions.

The current study provides a comprehensive look of the adsorption process of Candida rugosa lipase (CRL) on Ca2Fe2O5 iron oxide nanoparticles (NPs). Protein-support interactions were identified across a broad range of pH and ionic strengths (mM) through a response surface methodology, surface charge determination, and spectroscopic and in silico analyses. The maximum quantity of immobilized protein was achieved at an ionic strength of 50 mM and pH 4. However, this condition did not allow for the greatest hydrolytic activity to be obtained. Indeed, it was recorded at acidic pH, but at 150 mM, where evaluation of the recovered activity revealed hyperactivation of the enzyme. These findings were supported by adsorption isotherms performed under different conditions. Based on zeta potential measurements, electrostatic interactions contributed differently to protein-support binding under the conditions tested, showing a strong correlation with experimentally determined immobilization parameters. Raman spectra revealed an increase in hydrophobicity around tryptophan residues, whereas the enzyme immobilization significantly reduced the phenylalanine signal in CRL. This suggests that this residue was involved in the interaction with Ca2Fe2O2 and molecular docking analysis confirmed these findings. Fluorescence spectroscopy showed distinct behaviors in the CRL emission patterns with the addition of Ca2Fe2O5 at pH 4 and 7. The calculated thermodynamic parameters indicated that the contact would be mediated by hydrophobic interactions at both pHs, as well as by ionic ones at pH 4. In this approach, this work adds to our understanding of the design of biocatalysts immobilized in iron oxide NPs.

Effect of cholesterol on the surface polarity and hydration of lipid interphases as measured by Laurdan fluorescence: New insights.

Comparison of the behavior of Laurdan in gel and in the liquid crystalline DPPC bilayers with that observed in chloroform and OctOH allow concluding that changes in the membrane lipid order cannot be ascribed to changes in viscosity of the local environment. Cholesterol acts as a spacer below the transition temperature of DPPC, promoting a disorder state in the acyl chain region. No evidence of water entrance has been detected with Laurdan up to 30% Cholesterol in DPPC in this condition. In contrast, Chol displaces to longer values the wavelength of Laurdan in membranes in the liquid crystalline state. This decrease in polarity occurs above 5% Chol and is directly related to the water extrusion produced by Chol. This effect is similar to that occurring in liquid crystalline membranes subjected to hypertonic stress. The behavior is comparable to that of Laurdan in OctOH at different water ratios below 5% Chol/DPPC. At higher ratios, other changes are evident.

Hepatic miR-33a/miR-144 and their target gene ABCA1 are associated with steatohepatitis in morbidly obese subjects.

BACKGROUND AND AIM: Abnormal cholesterol metabolism may contribute to the pathogenesis of non-alcoholic steatohepatitis (NASH) and fibrosis. miR-33 and miR-144 regulate adenosine triphosphate binding cassette transporter (ABCA1) and other target genes involved in cholesterol efflux, fatty acid oxidation and inflammation. We explored relationships between non-alcoholic fatty liver disease (NAFLD) and the hepatic expression of ABCA1/ABCG1, as well as other target genes regulated by miR-33 (carnitine O-octanoyltransferase, CROT and hydroxyacyl-CoA-dehydrogenase ß-subunit, HADHB) and miR-144 (toll-like receptor-2, TLR2). Moreover, we evaluated whether the expression of these genes is correlated with miR-33a/b and miR-144 expression in Mexican individuals with morbid obesity. METHODS: Eighty-four morbidly obese subjects undergoing bariatric surgery were included in this study. Liver biopsies were obtained to measure hepatic triglyceride and free cholesterol contents, as well as ABCA1, ABCG1, CROT, HADHB, TLR2, miR-33a/b and miR-144 expression. RESULTS: Hepatic free cholesterol content was significantly increased in NASH as compared to non-NASH subjects, while ABCA1 and ABCG1 protein levels significantly decreased with NASH and fibrosis progression. The relative expression of miR-33a and miR-144 correlated inversely with ABCA1 but not with ABCG1 protein levels. Moreover, both miRNAs increased significantly in NASH individuals. miR-33 target genes CROT and HADHB correlated inversely with miR-33a. However, the expression of these genes was not associated with NASH. CONCLUSIONS: miR-33a/144 and their target gene ABCA1 may contribute to the pathogenesis of NASH in morbidly obese subjects.

SFRP5 hepatic expression is associated with non-alcoholic liver disease in morbidly obese women.

BACKGROUND AND AIMS: Secreted frizzled-related protein 5 (SFRP5) was recently described as a new adipokine protective for hepatic steatosis and other obesity-related complications in the mouse model. To date, SFRP5 expression in non-alcoholic fatty liver disease (NAFLD) has not been fully assessed in humans. We measured circulating SFRP5 levels and its expression in liver and adipose tissue, and evaluated its association with NAFLD in morbidly obese women. MATERIAL AND METHODS: Fifty-four morbidly obese women undergoing bariatric surgery were included in the study. Liver biopsies were used for histology and hepatic triglyceride content quantification. Circulating SFRP5 levels were measured through enzyme-linked immunoabsorbent assay, and SFRP5 expression was performed in hepatic and adipose tissue (subcutaneous and visceral). RESULTS: Although circulating SFRP5 levels showed a tendency to decrease with NAFLD progression, no significant differences were observed among non-alcoholic steatosis, steatohepatitis, and control subjects. Hepatic SFRP5 expression showed a negative correlation with hepatic triglyceride content (r = -0.349, P = 0.016 for mRNA and r = -0.291, P = 0.040 for SRFP5 protein) and ALT serum levels (r = -0.437, P = 0.001 for SRFP5 protein). In addition, hepatic SFRP5 protein levels were significantly lower in NASH than in control subjects (P = 0.006). CONCLUSION: This is the first study reporting an association of hepatic SFRP5 expression with NAFLD in humans.

Tratamiento de la distrofia simpático refleja (DSR) con nitroglicerina transdérmica (Estudio descriptivo de 81 pacientes) / Treatment of the reflex sympathetic dystrophy (RSD) with transdermal nitroglycerin. Descriptive study on 81 patients

En 81 pacientes de DSR, estadio 1 predominante, se realizó en el período comprendido entre los años 1996-2000 un estudio descriptivo con base fisiopatológica con el objetivo de valorar la acción terapéutica de la nitroglicerina transdérmica en este síndrome. En la muestra estudiada se destacan: la edad media que fue de 51,5+15.2 años (rango 16 a 82 años), el sexo femenino en 61 casos (75,3 por ciento) y, entre los factores desencadenantes, los traumas: 50 casos (61,7 por ciento). El diagnóstico de DSR fue clínico, complementado por la centellografía ósea con 25 mC de Tc 99 DMP, realizada en 36 casos (44,4 por ciento), siendo positiva en 32 casos (89,0 por ciento), y por la radiografía de la zona afectada en 54 pacientes (66,6 por ciento), observándose osteoporosis difusa en 53 (98,1 por ciento). La nitroglicerina se utilizó bajo la forma farmacéutica de parches de absorción transdérmica a concentraciones de 25 y 50 mg con liberación horaria de 0 mg 2-0 mg 4. La duración del tratamiento promedio fue de 4,4+3,0 meses. La acción terapéutica clínica se demuestra por los resultados siguientes: como variables: la edad -años-, la evolución previa de la DSR -en meses- y tiempo de tratamiento en meses, en función de la respuesta al tratamiento (evolución excelente o muy buena o evolución buena o sin cambios). Un valor de p < 0,05 fue considerado para establecer la significación de los test empleados. Los resultados expresan que los valores de p son de 0,06, 0,004 y 0,04 respectivamente. Se demuestra, por ende, el valor terapéutico de la nitroglicerina principalmente cuando es suministrada precozmente. Los efectos secundarios adversos se manifestaron en 68 por ciento de los pacientes, que cedieron total o parcialmente, al continuar con la medicación y/o con la administración nocturna; un solo caso abandonó el estudio por intolerancia a la nitroglicerina.